Impaired immunoproteasomal function exacerbates renal ischemia-reperfusion injury.
Yasushi Ishii, Aya Fukui-Miyazaki, Sari Iwasaki, Takahiro Tsuji, Kiyohiko Hotta, Hajime Sasaki, Shimpei Nakagawa, Takuma Yoshida, Eri Murata, Koji Taniguchi, Nobuo Shinohara, Akihiro Ishizu, Masanori Kasahara, Utano Tomaru
Experimental and molecular pathology, 140, 104939, 104939, 18 Oct. 2024, [International Magazine]
English, Scientific journal, Oxidative stress caused by reactive oxygen species (ROS) is involved in the pathogenesis of renal ischemia-reperfusion injury (I/R injury), a major cause of acute kidney injury and delayed graft function (DGF). DGF is an early transplant complication that worsens graft prognosis and patient survival, but the underlying molecular changes are unclear. The proteasome is a multicatalytic enzyme complex that degrades both normal and damaged proteins, and recent studies have revealed that the immunoproteasome, a specific proteasome isoform whose proteolytic activity enhances the generation of antigenic peptides, plays critical roles in the cellular response against oxidative stress. In this study, we demonstrate the impact of the immunoproteasome in human DGF and in a mouse model of I/R injury. In patients with DGF, the expression of β5i, a specific immunoproteasome subunit, was decreased in vascular endothelial cells. In a mouse model, β5i knockout (KO) exacerbated renal I/R injury. KO mice showed greater inflammation, oxidative stress, and endothelial damage compared with wild-type mice. Impaired immunoproteasomal activity also caused increased cell death, ROS production, and expression of inflammatory factors in mouse renal vascular endothelial cells under conditions of hypoxia and reoxygenation. In conclusion, reduced expression of the immunoproteasomal catalytic subunit β5i exacerbates renal I/R injury in vivo, potentially increasing the risk of DGF. Further research targeting β5i expression in DGF could lead to the development of novel therapeutic strategies and biomarkers.

A Novel Ultrasound Finding for the Diagnosis of Giant Cell Arteritis: Comparison With Temporal Artery Biopsy Findings.
Yusuke Kudo, Karin Hara, Michito Murayama, Sanae Kaga, Satomi Omotehara, Takahito Iwai, Masaru Kato, Utano Tomaru, Yoshihiro Matsuno, Akihiro Ishizu
Journal of clinical ultrasound : JCU, 12 Oct. 2024, [International Magazine]
English, Scientific journal, Hypoechoic halo is a typical ultrasound finding in giant cell arteritis (GCA), but it may be a false positive due to arteriosclerosis. Therefore, we focused on the segmental distribution of GCA lesions, defined the luminal irregularity in long-axis images as the string of beads sign, and examined its diagnostic ability. As a result, the C-statistic of hypoechoic halo and string of beads sign was better than that of hypoechoic halo alone (1.00 vs. 0.89). Based on the above, the diagnostic ability of GCA can be improved by adding the string of beads sign to the conventional hypoechoic halo.

Cathepsin C inhibition reduces neutrophil serine protease activity and improves activated neutrophil-mediated disorders.
Yuka Nishibata, Suishin Arai, Mai Taniguchi, Issei Nakade, Hodaka Ogawa, Shota Kitano, Yumeka Hosoi, Ayano Shindo, Ryo Nishiyama, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Takafumi Shimizu, William Sinko, Tadashi Nagakura, Yoh Terada, Akihiro Ishizu
Nature communications, 15, 1, 6519, 6519, 22 Aug. 2024, [International Magazine]
English, Scientific journal, Cathepsin C (CatC) is an enzyme which regulates the maturation of neutrophil serine proteases (NSPs) essential for neutrophil activation. Activated neutrophils are key players in the innate immune system, and are also implicated in the etiology of various inflammatory diseases. This study aims to demonstrate a therapeutic potential for CatC inhibitors against disorders in which activated neutrophil-derived neutrophil extracellular traps (NETs) play a significant role. We demonstrate that a CatC inhibitor, MOD06051, dose-dependently suppresses the cellular activity of NSPs, including neutrophil elastase (NE), in vitro. Neutrophils derived from MOD06051-administered rats exhibit significantly lower NE activity and NET-forming ability than controls. Furthermore, MOD06051 dose-dependently ameliorates vasculitis and significantly decreases NETs when administered to a rat model of myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody-associated vasculitis (AAV). These findings suggest that CatC inhibition is a promising strategy to reduce neutrophil activation and improve activated neutrophil-mediated diseases such as MPO-AAV.

Decreased proteasome function increases oxidative stress in the early stage of pressure ulcer development.
Eri Murata, Takuma Yoshida, Utano Tomaru, Saaki Yamamoto, Aya Fukui-Miyazaki, Akihiro Ishizu, Masanori Kasahara
Experimental and molecular pathology, 137, 104891, 104891, Jun. 2024, [International Magazine]
English, Scientific journal, The aging process in the elderly results in heightened skin fragility associated with various disorders, including pressure ulcers (PUs). Despite the high incidence of PUs in the elderly population, there is a limited body of research specifically examining the impact of aging on the development of pressure ulcers. Therefore, investigating age-related physiological abnormalities is essential to elucidate the pathogenesis of PUs. Ischemia-reperfusion (I/R) injury and the subsequent oxidative stress caused by reactive oxygen species (ROS) play essential roles in the early stage of PUs. In this study, we used a mouse model of proteasomal dysfunction with an age-related phenotype to examine the role of proteasome activity in cutaneous I/R injury in vivo. Decreased proteasome function did not affect the expression of inflammatory cytokines and adhesion molecules in the I/R area in transgenic mice; however, proteasome inhibition increased oxidative stress that was not attenuated by activation of the oxidative stress response mediated by NF-E2-related factor 2 (Nrf2). In dermal fibroblasts (FCs) subjected to hypoxia-reoxygenation (H/R), proteasome inhibition induced oxidative stress and ROS production, and Nrf2 activation did not adequately upregulate antioxidant enzyme expression, possibly leading to antioxidant/oxidant imbalance. The free radical scavenger edaravone had protective effects against I/R injury in vivo and decreased oxidative stress in FCs treated with a proteasome inhibitor and subjected to H/R in vitro. The results suggest that the age-related decline in proteasome activity promotes cutaneous I/R injury-induced oxidative stress, and free radical scavengers may exert protective effects by preventing oxidative stress in the early stage of PUs.

Decreased Proteasomal Function Exacerbates Endoplasmic Reticulum Stress-Induced Chronic Inflammation in Obese Adipose Tissue.
Shimpei Nakagawa, Aya Fukui-Miyazaki, Takuma Yoshida, Yasushi Ishii, Eri Murata, Koji Taniguchi, Akihiro Ishizu, Masanori Kasahara, Utano Tomaru
The American journal of pathology, 194, 6, 1033, 1046, Jun. 2024, [International Magazine]
English, Scientific journal, Low-grade chronic inflammation contributes to both aging and the pathogenesis of age-related diseases. White adipose tissue (WAT) in obese individuals exhibits chronic inflammation, which is associated with obesity-related disorders. Aging exacerbates obesity-related inflammation in WAT; however, the molecular mechanisms underlying chronic inflammation and its exacerbation by aging remain unclear. Age-related decline in activity of the proteasome, a multisubunit proteolytic complex, has been implicated in age-related diseases. This study employed a mouse model with decreased proteasomal function that exhibits age-related phenotypes to investigate the impact of adipocyte senescence on WAT inflammation. Transgenic mice expressing proteasomal subunit β5t with weak chymotrypsin-like activity experience reduced lifespan and develop age-related phenotypes. Mice fed with a high-fat diet and experiencing proteasomal dysfunction exhibited increased WAT inflammation, increased infiltration of proinflammatory M1-like macrophages, and increased proinflammatory adipocytokine-like monocyte chemoattractant protein-1, plasminogen activator inhibitor-1, and tumor necrosis factor-α, which are all associated with activation of endoplasmic reticulum (ER) stress-related pathways. Impaired proteasomal activity also activated ER stress-related molecules and induced expression of proinflammatory adipocytokines in adipocyte-like cells differentiated from 3T3-L1 cells. Collectively, the results suggesed that impaired proteasomal activity increases ER stress and that subsequent inflammatory pathways play pivotal roles in WAT inflammation. Because proteasomal function declines with age, age-related proteasome impairment may be involved in obesity-related inflammation among elderly individuals.

Transcription factor Nrf2 activation regulates NETosis, endothelial injury, and kidney disease in myeloperoxidase-positive antineutrophil cytoplasmic antibody-associated vasculitis.
Yusho Ueda, Daigo Nakazawa, Saori Nishio, Satoka Shiratori-Aso, Takashi Kudo, Atsuko Miyoshi-Harashima, Kanako Watanabe-Kusunoki, Fumihiko Hattanda, Sari Iwasaki, Takahiro Tsuji, Utano Tomaru, Yasuaki Aratani, Mamiko Yamamoto, Akihiro Ishizu, Tatsuya Atsumi
Kidney international, 25 Mar. 2024, [International Magazine]
English, Scientific journal, Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a systemic autoimmune disease pathologically characterized by vascular necrosis with inflammation. During AAV development, activated neutrophils produce reactive oxygen species (ROS), leading to the aberrant formation of neutrophil extracellular traps (NETs) via NETosis and subsequent fibrinoid vascular necrosis. Nuclear factor-erythroid 2-related factor 2 (Nrf2) functions as an intracellular defense system to counteract oxidative stress by providing antioxidant properties. Herein, we explored the role of Nrf2 in the pathogenesis of AAV. The role and mechanism of Nrf2 in ANCA-stimulated neutrophils and subsequent endothelial injury were evaluated in vitro using Nrf2 genetic deletion and Nrf2 activator treatment. In corresponding in vivo studies, the role of Nrf2 in ANCA-transfer AAV and spontaneous AAV murine models was examined. Pharmacological activation of Nrf2 in vitro suppressed ANCA-induced NET formation via the inhibition of ROS. In contrast, NET formation was enhanced in Nrf2-deficient neutrophils. Furthermore, Nrf2 activation protected endothelial cells from ANC-induced NETs-mediated injury. In vivo, Nrf2 activation ameliorated glomerulonephritis in two AAV models by upregulating antioxidants and inhibiting ROS-mediated NETs. Furthermore, Nrf2 activation restrained the expansion of splenic immune cells, including T lymphocytes and limited the infiltration of Th17 cells into the kidney. In contrast, Nrf2 genetic deficiency exacerbated vasculitis in a spontaneous AAV model. Thus, the pathophysiological process in AAV may be downregulated by Nrf2 activation, potentially leading to a new therapeutic strategy by regulating NETosis.

Autophagy inhibition-induced cytosolic DNA sensing combined with differentiation therapy induces irreversible myeloid differentiation in leukemia cells.
Tomohisa Baba, Utano Tomaru, Atsushi Hirao, Naofumi Mukaida, Yoshikazu Johmura
Cancer research communications, 11 Mar. 2024, [International Magazine]
English, Scientific journal, Accumulating evidence indicates that various oncogenic mutations interfere with normal myeloid differentiation of leukemogenic cells during the early process of acute myeloid leukemia (AML) development. Differentiation therapy is a therapeutic strategy capable of terminating leukemic expansion by reactivating the differentiation potential; however, the plasticity and instability of leukemia cells counteract the establishment of treatments aimed at irreversibly inducing and maintaining their differentiation states. Based on our previous observation that autophagy inhibitor treatment induces the accumulation of cytosolic DNA and activation of cytosolic DNA-sensor signaling selectively in leukemia cells, we herein examined the synergistic effect of cytosolic DNA-sensor signaling activation with conventional differentiation therapy on AML. The combined treatment succeeded in inducing irreversible differentiation in AML cell lines. Mechanistically, cytosolic DNA was sensed by absent in melanoma 2 (AIM2), a cytosolic DNA sensor. Activation of the AIM2 inflammasome resulted in the accumulation of p21 through the inhibition of its proteasomal degradation, thereby facilitating the myeloid differentiation. Importantly, the combined therapy dramatically reduced the total leukemia cell counts and proportion of blast cells in the spleens of AML mice. Collectively, these findings indicate that the autophagy inhibition-cytosolic DNA-sensor signaling axis can potentiate AML differentiation therapy.

Methylprednisolone pulse-enhanced neutrophil extracellular trap formation in mice with imiquimod-induced lupus-like disease, resulting in ischaemia of the femoral head cartilage.
Hodaka Ogawa, Shunichi Yokota, Yumeka Hosoi, Ayano Shindo, Naho Ogawa, Ryodai Yamamura, Tomohiro Shimizu, Issei Nakade, Suishin Arai, Mai Taniguchi, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Norimasa Iwasaki, Akihiro Ishizu
Lupus science & medicine, 10, 2, 28 Dec. 2023, [International Magazine]
English, Scientific journal, OBJECTIVES: Methylprednisolone (mPSL) pulse therapy is an essential option for patients with active systemic lupus erythematosus, but there is a risk of adverse events related to microcirculation disorders, including idiopathic osteonecrosis of the femoral head (ONFH). Recent studies have revealed that excessive neutrophil extracellular traps (NETs) are involved in microcirculation disorders. This study aimed to demonstrate that mPSL pulse could induce NETs in lupus mice and identify the factors contributing to this induction. METHODS: Six mice with imiquimod (IMQ)-induced lupus-like disease and six normal mice were intraperitoneally injected with mPSL on days 39 to 41, and five mice with IMQ-induced lupus-like disease and six normal mice were injected with phosphate-buffered saline. Pathological examinations were conducted to evaluate the ischaemic state of the femoral head and tissue infiltration of NET-forming neutrophils. Proteome analysis was performed to extract plasma proteins specifically elevated in mPSL-administered mice with IMQ-induced lupus-like disease, and their effects on NET formation were assessed in vitro. RESULTS: Mice with IMQ-induced lupus-like disease that received mPSL pulse demonstrated ischaemia of the femoral head cartilage with tissue infiltration of NET-forming neutrophils. Proteome analysis suggested that prenylcysteine oxidase 1 (PCYOX1) played a role in this phenomenon. The reaction of PCYOX1-containing very low-density lipoproteins (VLDL) with its substrate farnesylcysteine (FC) induced NETs in vitro. The combined addition of IMQ and mPSL synergistically enhanced VLDL-plus-FC-induced NET formation. CONCLUSION: PCYOX1 and related factors are worthy of attention to understand the underlying mechanisms and create novel therapeutic strategies for mPSL-mediated microcirculation disorders, including ONFH.

Bruton's tyrosine kinase is a possible therapeutic target in microscopic polyangiitis.
Issei Nakade, Yuto Tamura, Fuyu Hashimoto, Yuko Ariza, Shingo Hotta, Hirofumi Fujigaya, Suishin Arai, Mai Taniguchi, Hodaka Ogawa, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
Arthritis research & therapy, 25, 1, 215, 215, 06 Nov. 2023, [International Magazine]
English, Scientific journal, BACKGROUND: Bruton's tyrosine kinase (Btk) is an enzyme expressed in leukocytes other than T lymphocytes and plasma cells and involved in B-cell receptor- and Fcγ receptor (FcγR)-mediated signal transduction. Btk inhibitors potentially suppress autoantibody production due to the expected inhibitory ability of B lymphocyte differentiation into antibody-producing plasma cells and reduce FcγR-mediated neutrophil activation, including the release of neutrophil extracellular traps (NETs). Microscopic polyangiitis (MPA) is a systemic small-vessel vasculitis characterized by the pathogenic autoantibody, antineutrophil cytoplasmic antibody (ANCA) that reacts with myeloperoxidase (MPO). MPO and MPO-ANCA immune complex (IC)-induced FcγR-mediated NETs are critically involved in MPA pathogenesis. This study aimed to demonstrate the therapeutic efficacy of the Btk inhibitor tirabrutinib on MPA. METHODS: Various doses of tirabrutinib or vehicle were orally administered to Sprague-Dawley rats daily. Four weeks later, the number of peripheral B lymphocytes was counted, and Btk phosphorylation in B lymphocytes was evaluated by flow cytometry. Human peripheral blood neutrophils were stimulated by MPO and anti-MPO antibody ICs (MPO and anti-MPO-ICs), and Btk and its downstream Vav phosphorylation were assessed by western blotting. The effects of tirabrutinib on MPO and anti-MPO-IC-induced NET formation were examined in vitro. Wistar Kyoto rats were immunized with human MPO to induce experimental MPA and given drug-free or tirabrutinib-containing feed (0.0037% or 0.012%) from day 0 or 28. All rats were euthanized on day 42 for serological and histological evaluation. RESULTS: Tirabrutinib inhibited Btk phosphorylation without decreasing B lymphocytes in vivo. Neutrophil Btk and Vav were phosphorylated when stimulated with MPO and anti-MPO-ICs. Tirabrutinib suppressed MPO and anti-MPO-IC-induced NET formation in vitro and ameliorated experimental MPA in a dose-dependent manner in vivo. Although MPO-ANCA production was not affected, NET-forming neutrophils in the blood were significantly reduced by tirabrutinib. CONCLUSIONS: The Btk inhibitor tirabrutinib suppressed MPO and anti-MPO-IC-induced NET formation in vitro and ameliorated experimental MPA by reducing NET-forming neutrophils but not decreasing MPO-ANCA titer in vivo. This study suggests that Btk is a possible therapeutic target in MPA.

Inhibition of Toll-like receptor 4 and Interleukin-1 receptor prevent SARS-CoV-2 mediated kidney injury.
Daigo Nakazawa, Yohei Takeda, Masatoshi Kanda, Utano Tomaru, Haruko Ogawa, Takashi Kudo, Satoka Shiratori-Aso, Kanako Watanabe-Kusunoki, Yusho Ueda, Atsuko Miyoshi, Fumihiko Hattanda, Saori Nishio, Ryo Uozumi, Akihiro Ishizu, Tatsuya Atsumi
Cell death discovery, 9, 1, 293, 293, 10 Aug. 2023, [International Magazine]
English, Scientific journal, Acute kidney injury (AKI) is a common and severe complication of the coronavirus disease 2019 (COVID-19). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) directly affects the glomerular and tubular epithelial cells to induce AKI; however, its pathophysiology remains unclear. Here, we explored the underlying mechanisms and therapeutic targets of renal involvement in COVID-19. We developed an in vitro human kidney cellular model, including immortalized tubular epithelial and endothelial cell lines, demonstrating that SARS-CoV-2 directly triggers cell death. To identify the molecular targets in the process of SARS-CoV-2-mediated cell injury, we performed transcriptional analysis using RNA sequencing. Tubular epithelial cells were more prone to dying by SARS-CoV-2 than endothelial cells; however, SARS-CoV-2 did not replicate in renal cells, distinct from VeroE6/transmembrane protease serine 2 cells. Transcriptomic analysis revealed increased inflammatory and immune-related gene expression levels in renal cells incubated with SARS-CoV-2. Toll-like receptor (TLR) 3 in renal cells recognized viral RNA and underwent cell death. Furthermore, analysis of upstream regulators identified several key transcriptional regulators. Among them, inhibition of the interleukin-1 receptor (IL-1R) and TLR4 pathways protects tubular epithelial and endothelial cells from injury via regulation of the signal transducer and activator of transcription protein-3/nuclear factor-kB pathway. Our results reveal that SARS-CoV-2 directly injures renal cells via the proinflammatory response without viral replication, and that IL-1R and TLR4 may be used as therapeutic targets for SARS-CoV-2 mediated kidney injury.

CD47 blockade ameliorates autoimmune vasculitis via efferocytosis of neutrophil extracellular traps.
Satoka Shiratori-Aso, Daigo Nakazawa, Takashi Kudo, Masatoshi Kanda, Yusho Ueda, Kanako Watanabe-Kusunoki, Saori Nishio, Sari Iwasaki, Takahiro Tsuji, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu, Tatsuya Atsumi
JCI insight, 27 Jun. 2023, [International Magazine]
English, Scientific journal, Neutrophil extracellular trap (NET) formation contributes to immune defense and is a distinct form of cell death. Excessive NET formation is found in patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), contributing to disease progression. The clearance of dead cells by macrophages, a process known as efferocytosis, is regulated by the CD47-mediated "don't eat me" signal. Hence, we hypothesized that pathogenic NETs in AAV escape from efferocytosis via the CD47 signaling pathway, resulting in the development of necrotizing vasculitis. Immunostaining for CD47 in human renal tissues revealed high CD47 expression in crescentic glomerular lesions of patients with AAV. In ex vivo studies, ANCA-induced netting neutrophils increased the expression of CD47 with the reduction of efferocytosis. After efferocytosis, macrophages displayed pro-inflammatory phenotypes. The blockade of CD47 in spontaneous crescentic glomerulonephritis-forming/Kinjoh (SCG/Kj) mice ameliorated renal disease and reduced myeloperoxidase (MPO)-ANCA titers with a reduction in NETs formation. Thus, CD47 blockade would protect against developing glomerulonephritis in AAV via restored efferocytosis of ANCA-induced NETs.

EGFR変異陽性肺癌におけるCD24発現の検討　　　　　　　　　　　　　　　
椎谷 研彦, 野口 卓郎, 外丸 詩野, 有賀 伸, 高島 雄太, 品川 尚文, 木下 一郎, 松野 吉宏, 榊原 純, 秋田 弘俊
日本病理学会会誌, 112, 1, 247, 247, (一社)日本病理学会, Mar. 2023
Japanese

ANCA関連血管炎に対するアバコパンを含む新規治療 C5a受容体拮抗薬と好中球エラスターゼ阻害剤の好中球活性化抑制比較　　　　　　　　　　　　　　　
荒井 粋心, 西端 友香, 谷口 舞, 益田 紗季子, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 629, 629, (一社)日本リウマチ学会, Mar. 2023
Japanese

ANCA関連血管炎に対するアバコパンを含む新規治療 Cathepsin C阻害による好中球細胞外トラップ形成抑制　　　　　　　　　　　　　　　
西端 友香, 荒井 粋心, 谷口 舞, 益田 紗季子, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 629, 629, (一社)日本リウマチ学会, Mar. 2023
Japanese

ANCA関連血管炎に対するアバコパンを含む新規治療 好中球細胞外トラップにDNase I抵抗性を付与するタンパクの同定　　　　　　　　　　　　　　　
谷口 舞, 益田 紗季子, 中村 哲朗, 荒井 粋心, 西端 友香, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 630, 630, (一社)日本リウマチ学会, Mar. 2023
Japanese

ベーチェット病 ベーチェット病における口内炎の発生原因の解明　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 川邊 智宏, 宮前 多佳子, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 649, 649, (一社)日本リウマチ学会, Mar. 2023
Japanese

自己抗体から紐解く疾患の病理病態 ANCA関連血管炎におけるintermolecular epitope spreadingによる抗GBM抗体の産生　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
日本病理学会会誌, 112, 1, 196, 196, (一社)日本病理学会, Mar. 2023
Japanese

SLEモデルマウスへのステロイドパルスは好中球細胞外トラップを誘導する　　　　　　　　　　　　　　　
小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 886, 886, (一社)日本リウマチ学会, Mar. 2023
Japanese

NKG2D Ligand Expression Induced by Oxidative Stress Mitigates Cutaneous Ischemia-Reperfusion Injury.
Keishi Makita, Noriyuki Otsuka, Utano Tomaru, Koji Taniguchi, Masanori Kasahara
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 71, 2, 61, 72, Feb. 2023, [International Magazine]
English, Scientific journal, Pressure ulcers represent a crucial clinical problem, especially in hospitalized patients. Ischemia-reperfusion (I-R) is an important cause of these lesions. Natural killer (NK), invariant NK T (iNKT), and dendritic epidermal T-cells, which express the natural killer group 2, member D (NKG2D) receptor, have been reported to have physiological roles in skin tissue repair and wound healing. However, a role for NKG2D-NKG2D ligand interactions in I-R-induced skin injury has not been determined. Using a murine pressure ulcer model, we demonstrated that I-R-induced ulcers in NKG2D-deficient mice were larger than those in wild-type or T-cell receptor δ knockout mice. Histopathological evaluation revealed that accumulation of macrophages and neutrophils at the peripheral deep dermis and subcutaneous tissue of the ulcers was enhanced in NKG2D-deficient mice. Rae-1 mRNA, which encodes an NKG2D ligand, was induced, and RAE-1 protein was detected immunohistochemically in fibroblasts and inflammatory cells in the dermis after reperfusion. RAE-1 expression was also increased in primary mouse fibroblasts treated with sodium arsenite. These results suggested that NKG2D ligand expression was induced by oxidative stress after I-R injury and support a putative role for this ligand in wound repair. Furthermore, the influx of NKG2D-positive cells at I-R sites may mitigate pressure ulcers via NKG2D-NKG2D ligand interactions.

巨細胞性動脈炎の診断に寄与する新たな超音波所見 生検所見との比較に基づく検討　　　　　　　　　　　　　　　
工藤 悠輔, 原 花梨, 村山 迪史, 加賀 早苗, 表原 里実, 岩井 孝仁, 進藤 由衣香, 菊池 桃佳, 加藤 将, 外丸 詩野, 松野 吉宏, 石津 明洋
脈管学, 63, 1, 16, 16, (一社)日本脈管学会, Feb. 2023
Japanese

COVID-19関連を含むIgA血管炎皮膚生検標本を使用したNeutrophil Extracellular Traps(NETs)の検証　　　　　　　　　　　　　　　
川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 石津 明洋, 外丸 詩野
日本皮膚科学会雑誌, 133, 2, 254, 254, (公社)日本皮膚科学会, Feb. 2023
Japanese

感染症と血管炎 COVID-19発症後およびCOVID-19ワクチン接種後IgA血管炎の皮膚生検組織における好中球細胞外トラップの沈着 COVID-19非関連IgA血管炎との比較　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 外丸 詩野, 横山 華英, 池田 高治, 川上 民裕, 石津 明洋
脈管学, 63, 1, 10, 11, (一社)日本脈管学会, Feb. 2023
Japanese

無症候性血尿を呈した抗糸球体基底膜(GBM)抗体陽性症例の血清を用いた抗体解析　　　　　　　　　　　　　　　
西端 友香, 佐藤 雅之, 長森 恒久, 益田 紗季子, 外丸 詩野, 石津 明洋
脈管学, 63, 1, 13, 14, (一社)日本脈管学会, Feb. 2023
Japanese

皮膚血管炎動物モデルの完成　　　　　　　　　　　　　　　
川上 民裕, 中出 一生, 田村 宥人, 伊藤 吹夕, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
脈管学, 63, 1, 15, 15, (一社)日本脈管学会, Feb. 2023
Japanese

巨細胞性動脈炎の診断に寄与する新たな超音波所見 生検所見との比較に基づく検討　　　　　　　　　　　　　　　
工藤 悠輔, 原 花梨, 村山 迪史, 加賀 早苗, 表原 里実, 岩井 孝仁, 進藤 由衣香, 菊池 桃佳, 加藤 将, 外丸 詩野, 松野 吉宏, 石津 明洋
脈管学, 63, 1, 16, 16, (一社)日本脈管学会, Feb. 2023
Japanese

EGFR inhibition in EGFR-mutant lung cancer cells perturbs innate immune signaling pathways in the tumor microenvironment.
Akihiko Shiiya, Takuro Noguchi, Utano Tomaru, Shin Ariga, Yuta Takashima, Yoshihito Ohhara, Jun Taguchi, Satoshi Takeuchi, Yasushi Shimizu, Ichiro Kinoshita, Tomonobu Koizumi, Yoshihiro Matsuno, Naofumi Shinagawa, Jun Sakakibara-Konishi, Hirotoshi Dosaka-Akita
Cancer science, 18 Dec. 2022, [International Magazine]
English, Scientific journal, Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) elicit potent cell cycle arrest in EGFR-mutant non-small-cell lung cancer (NSCLC) cells. However, little is known about the mechanisms through which these drugs alter the tumor phenotype that contributes to the immune escape of EGFR-mutant cells. Using EGFR-mutant NSCLC cell lines and tissue samples from patients, we investigated the changes in immune checkpoints expressed in tumor cells following EGFR inhibition. Subsequently, we also analyzed the role of soluble factors from the dying tumor cells in the activation of immune signaling pathways involved in therapy resistance. Upon EGFR-TKI treatment, we found that EGFR-mutant cells upregulated the expression of innate immune checkpoint CD24 in vitro. We then analyzed biopsy samples from six patients who developed resistance to a first-generation EGFR-TKI without the acquired T790M mutation. Immunohistochemistry revealed that levels of tumor CD24 expression were increased upon treatment compared with those from pre-treatment samples. Monocyte-derived macrophages facilitated antibody-dependent cellular phagocytosis when EGFR-TKI-treated EGFR-mutant cells were incubated with anti-CD24 antibodies in vitro, suggesting that CD24 may be a therapeutical target for EGFR-mutant lung cancer. Moreover, EGFR inhibition accelerated the release of cell-free DNA (cfDNA) from dying tumor cells, which activated the type I interferon signaling pathways in human THP-1 monocytes in a stimulator of interferon genes-dependent manner. Our study indicates that EGFR inhibition in EGFR-mutant NSCLC cells fosters a tumor microenvironment associated with immune escape. Thus, CD24 targeted therapy and cfDNA monitoring may contribute to improved treatment outcomes in patients with EGFR-mutant NSCLC.

Low disease activity of microscopic polyangiitis in patients with anti-myosin light chain 6 antibody that disrupts actin rearrangement necessary for neutrophil extracellular trap formation.
Miku Yoshinari, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Yoshihiro Arimura, Koichi Amano, Yukio Yuzawa, Ken-Ei Sada, Tatsuya Atsumi, Hiroaki Dobashi, Hitoshi Hasegawa, Masayoshi Harigai, Seiichi Matsuo, Hirofumi Makino, Akihiro Ishizu
Arthritis research & therapy, 24, 1, 274, 274, 16 Dec. 2022, [International Magazine]
English, Scientific journal, BACKGROUND: Neutrophil extracellular traps (NETs) are critically involved in microscopic polyangiitis (MPA) pathogenesis, and some patients with MPA possess anti-NET antibody (ANETA). Anti-myosin light chain 6 (MYL6) antibody is an ANETA that affects NETs. This study aimed to determine the significance of anti-MYL6 antibody in MPA. METHODS: The influence of anti-MYL6 antibody on NET formation and actin rearrangement necessary for NET formation was assessed by fluorescent staining. An enzyme-linked immunosorbent assay was established to detect serum anti-MYL6 antibody, and the prevalence of this antibody in MPA was determined. Furthermore, the disease activity and response to remission-induction therapy of MPA were compared between anti-MYL6 antibody-positive and anti-MYL6 antibody-negative MPA patients. RESULTS: Anti-MYL6 antibody disrupted G-actin polymerization into F-actin, suppressing phorbol 12-myristate 13-acetate-induced NET formation. Serum anti-MYL6 antibody was detected in 7 of 59 patients with MPA. The Birmingham vasculitis activity score (BVAS) of anti-MYL6 antibody-positive MPA patients was significantly lower than anti-MYL6 antibody-negative MPA patients. Among the nine BVAS evaluation items, the cutaneous, cardiovascular, and nervous system scores of anti-MYL6 antibody-positive MPA patients were significantly lower than anti-MYL6 antibody-negative MPA patients, although other items, including the renal and chest scores, were equivalent between the two groups. The proportion of patients with remission 6 months after initiation of remission-induction therapy in anti-MYL6 antibody-positive MPA patients was significantly higher than in anti-MYL6 antibody-negative MPA patients. CONCLUSIONS: Collective findings suggested that anti-MYL6 antibody disrupted actin rearrangement necessary for NET formation and could reduce the disease activity of MPA.

Similar deposition of neutrophil extracellular traps in the dermis among COVID-19-associated IgA vasculitis, post-COVID-19 vaccination IgA vasculitis, and COVID-19-unrelated IgA vasculitis.
Tamihiro Kawakami, Kae Yokoyama, Takaharu Ikeda, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
The Journal of dermatology, 50, 5, e151-e152, 15 Dec. 2022, [International Magazine]
English

正常ラットにヒストン皮下注射後,抗ホスファチジルセリン・プロトロンビン複合体抗体と抗リソソーム膜タンパク質2抗体の静脈注射により,皮膚血管炎の発症に成功した　　　　　　　　　　　　　　　
川上 民裕, 中出 一生, 田村 宥人, 伊藤 吹夕, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
日本皮膚免疫アレルギー学会総会学術大会プログラム・抄録集, 52回, 215, 215, (一社)日本皮膚免疫アレルギー学会, Dec. 2022
Japanese

正常ラットにヒストン皮下注射後,抗ホスファチジルセリン・プロトロンビン複合体抗体と抗リソソーム膜タンパク質2抗体の静脈注射により,皮膚血管炎の発症に成功した　　　　　　　　　　　　　　　
川上 民裕, 中出 一生, 田村 宥人, 伊藤 吹夕, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
日本皮膚免疫アレルギー学会総会学術大会プログラム・抄録集, 52回, 215, 215, (一社)日本皮膚免疫アレルギー学会, Dec. 2022
Japanese

A Novel Antineutrophil Extracellular Trap Antibody Targeting Myosin Light Chain 6 in Microscopic Polyangiitis.
Miku Yoshinari, Fumihiko Hattanda, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
The Journal of rheumatology, 49, 11, 1286, 1288, Nov. 2022, [International Magazine]
English

Transcriptional dynamics of granulocytes in direct response to incubation with SARS-CoV-2.
Daigo Nakazawa, Yohei Takeda, Masatoshi Kanda, Utano Tomaru, Haruko Ogawa, Takashi Kudo, Satoka Shiratori-Aso, Kanako Watanabe-Kusunoki, Yusho Ueda, Atsuko Miyoshi, Fumihiko Hattanda, Saori Nishio, Ryo Uozumi, Akihiro Ishizu, Tatsuya Atsumi
FEBS open bio, 22 Oct. 2022, [International Magazine]
English, Scientific journal, Severe coronavirus disease 2019 (COVID-19) is characterized by acute respiratory distress syndrome and multiple organ dysfunction, in which the host immune response plays a pivotal role. Excessive neutrophil activation and subsequent superfluity of neutrophil extracellular traps (NETs) can lead to tissue damage, and several studies have shown the involvement of neutrophils in severe COVID-19. However, the detailed responses of each neutrophil subset to SARS-CoV-2 infection has not been fully described. To explore this issue, we incubated normal-density granulocytes (NDGs) and low-density granulocytes (LDGs) with different viral titers of SARS-CoV-2. NDGs form NETs with chromatin fibers in response to SARS-CoV-2, whereas LDGs incubated with SARS-CoV-2 display a distinct morphology with condensed nuclei and moderate transcriptional changes. Based on these transcriptional changes, we suggest that AGO2 possibly plays a role in LDG regulation in response to SARS-CoV-2.

ベーチェット病皮下の血栓性静脈炎におけるNeutrophil Extracellular Trapsの発現
川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
アレルギー, 71, 6-7, 870, 870, (一社)日本アレルギー学会, Aug. 2022
Japanese

ベーチェット病皮膚生検標本を使用したNETsの検証　　　　　　　　　　　　　　　
川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
西日本皮膚科, 84, 4, 371, 371, 日本皮膚科学会-西部支部, Aug. 2022
Japanese

ベーチェット病皮下の血栓性静脈炎におけるNeutrophil Extracellular Trapsの発現
川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
アレルギー, 71, 6-7, 870, 870, (一社)日本アレルギー学会, Aug. 2022
Japanese

Cyclophilin D regulates NETosis and inflammation in myeloperoxidase-antineutrophil cytoplasmic antibody-associated vasculitis.
Takashi Kudo, Daigo Nakazawa, Kanako Watanabe-Kusunoki, Masatoshi Kanda, Satoka Shiratori-Aso, Nobuya Abe, Saori Nishio, Jun-Ichiro Koga, Sari Iwasaki, Takahiro Tsuji, Yuichiro Fukasawa, Miwako Yamasaki, Masahiko Watanabe, Sakiko Masuda, Utano Tomaru, Masaaki Murakami, Yasuaki Aratani, Akihiro Ishizu, Tatsuya Atsumi
Arthritis & rheumatology (Hoboken, N.J.), 75, 1, 71, 83, 29 Jul. 2022, [International Magazine]
English, Scientific journal, OBJECTIVE: Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is pathologically characterized by focal fibrinoid necrosis where ANCA-mediated neutrophil extracellular trap (NET) formation and subsequent endothelial necrosis occurs. Cyclophilin D (CypD) plays an important role in mediating cell necrosis and inflammation via opening of mitochondrial permeability transition pores (mPTP). Here, we examined the role of CypD in AAV pathogenesis. METHODS: In vitro, the role and mechanism of CypD in ANCA-stimulated neutrophils were assessed by immunostaining and electron microscopy. A comprehensive RNA sequencing analysis was performed on ANCA-treated murine neutrophils. To investigate the role of CypD in vivo, an-anti-MPO IgG-transfer AAV model or spontaneous AAV model mice were induced in CypD knockout or wild-type mice. RESULTS: In vitro, pharmacological and genetic inhibition of CypD suppressed ANCA-induced NET formation via the suppression of reactive oxygen species/cytochrome c release from the mitochondria. The analysis of RNA sequencing in ANCA-treated murine neutrophils revealed the involvement of inflammatory responses, and CypD deficiency reduced ANCA-induced alterations in gene expression. Furthermore, the upstream regulator analysis revealed the relevance of intracellular calcium (CypD activator) and cyclosporin (CypD inhibitor) in ANCA stimulation, indicating that CypD-dependent mPTP opening is associated with ANCA-induced neutrophil activation and NETosis. In both AAV models, the genetic deletion of CypD ameliorated crescentic glomerulonephritis via the inhibition of CypD-dependent neutrophil and endothelial necrosis. CONCLUSIONS: CypD targeting is a novel and specific therapeutic strategy for AAV via the resolution of necrotizing vasculitis.

Typical cutaneous small-vessel vasculitis induced by combined injection of antiphosphatidylserine/prothrombin complex antibody and anti-LAMP-2 antibody in normal rats.
Tamihiro Kawakami, Issei Nakade, Yuto Tamura, Fuyu Ito, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
The Journal of dermatology, 49, 12, 1233, 1237, 25 Jul. 2022, [International Magazine]
English, Scientific journal, We previously reported that IgA vasculitis and cutaneous arteritis could be dependently associated with the presence of the antiphosphatidylserine/prothrombin complex (anti-PS/PT) antibody and lysosomal-associated membrane protein-2 (LAMP-2). The copy number of LAMP-2 mRNA in skin tissue samples from rats with cutaneous vasculitis induced by intravenous administration of anti-PS/PT antibody after subcutaneous histone injection was significantly higher than in those without cutaneous vasculitis. We found LAMP-2 protein overexpression in neutrophils and vascular endothelial cells of the affected blood vessels in rats with cutaneous vasculitis induced by intravenous administration of anti-PS/PT antibody after cutaneous priming by histones. We found typical cutaneous small-vessel vasculitis in the skin of rats given intravenous injection of both anti-PS/PT antibody and anti-LAMP-2 antibody after cutaneous priming by histones. We suggested that the introduction of skin local histones and anti-PS/PT antibody in serum could move LAMP-2 to the cell surface of neutrophils and vascular endothelial cells, and that anti-LAMP-2 antibody could bridge these cells through antigen-specific binding in typical cutaneous small-vessel vasculitis.

Presence of neutrophil extracellular traps in superficial venous thrombosis of Behçet's disease.
Tamihiro Kawakami, Kae Yokoyama, Takaharu Ikeda, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
The Journal of dermatology, 49, 7, 741, 745, 17 Apr. 2022, [International Magazine]
English, Scientific journal, Behçet's disease (BD) has a heterogeneous spectrum of disease manifestations featuring the involvement of different organs and can be characterized with different symptoms depending on the clinical department in charge. We retrospectively reviewed BD patients seen at our hospital and investigated the presence of neutrophils producing neutrophil extracellular traps (NET) in those patients. Immunolabeling of myeloperoxidase and histone citrullination proteins was performed on skin biopsies from three BD patients who had skin biopsy-proven superficial vein thrombophlebitis in their erythema nodosum-like lesions. We observed a higher proportion of female patients and a higher incidence of acne-like eruptions among BD patients seen at our dermatology department, while there was a higher incidence of ocular and gastrointestinal involvement among BD patients treated in other departments. We suggest that sex statistical trends could lead to the co-development of different manifestations and may help clinicians choose the best therapeutic approaches, tailoring them to the specific phenotype of the patient rather than one based on single disease manifestations. NET were found in neutrophils of panniculitis concurrent with superficial vein thrombophlebitis. We suggest that the pathogenesis of BD-related thrombosis could be associated with neutrophil activation and NET are released in the panniculitis of affected skin lesions, erythema nodosum-like lesions.

肺病変にステロイドが奏功したPOIKTMPの1例(アンコール演題)
佐藤 理子, 鈴木 雅, 猪狩 智生, 中村 友彦, 高橋 桂, 佐々木 真知子, 木村 孔一, 木村 裕樹, 宮内 俊成, 乃村 俊史, 氏家 英之, 山口 直子, 大塚 紀幸, 外丸 詩野, 今野 哲
日本呼吸器学会誌, 11, 増刊, 272, 272, (一社)日本呼吸器学会, Apr. 2022
Japanese

Phorbol 12-myristate 13-acetate stimulation under hypoxia induces nuclear swelling with DNA outflow but not extracellular trap formation of neutrophils.
Sakiko Masuda, Kurumi Kato, Misato Ishibashi, Yuka Nishibata, Ayako Sugimoto, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Ichizo Tsujino, Akihiro Ishizu
Experimental and molecular pathology, 125, 104754, 104754, Apr. 2022, [International Magazine]
English, Scientific journal, Neutrophils stand sentinel over infection and possess diverse antimicrobial weapons, including neutrophil extracellular traps (NETs). NETs are composed of web-like extracellular DNA decorated with antimicrobial substances and can trap and eliminate invading microorganisms. Although phorbol 12-myristate 13-acetate (PMA) is a potent NET inducer, previous studies have demonstrated that not all neutrophils exhibit NET formation even if stimulated by PMA at high concentrations. This study first showed that some neutrophils stimulated by PMA displayed a swollen nucleus but not NET formation and that hypoxic environments suppressed the NET release. Next, characterization of PMA-stimulated neutrophils with a swollen nucleus was accomplished by differentiating between suicidal-type NETosis and apoptosis. Furthermore, the significance of the phenomenon was examined using formalin-fixed, paraffin-embedded human lung disease tissues with and without pneumonia. As a result, histone H3 citrullination, DNA outflow, propidium iodide labeling, resistance to DNase I, and suspended actin rearrangement were characteristics of PMA-stimulated neutrophils with a swollen nucleus distinct from neutrophils that underwent either suicidal-type NETosis or apoptosis. Neutrophils stimulated by PMA under hypoxic conditions secreted matrix metalloproteinase-9 cytotoxic to human lung-derived fibroblasts. Further, deposition of neutrophil-derived citrullinated histone H3+ chromatin substances in pulmonary lesions was greater in patients with pneumonia than in patients without pneumonia and positively correlated with hypoxia-inducible factor-1α expression. The collective findings suggested that neutrophils activated under hypoxic conditions could be putative modulators of hypoxia-related disease manifestations.

肺硬化性肺胞上皮腫および腺癌が同一結節内に併存した一例　　　　　　　　　　　　　　　
木内 隆之, 木内 静香, 大塚 紀幸, 外丸 詩野, 渡邊 幹夫, 松野 吉宏
日本病理学会会誌, 111, 1, 327, 327, (一社)日本病理学会, Mar. 2022
Japanese

好中球細胞外トラップにDNase I抵抗性を付与するタンパクの探索　　　　　　　　　　　　　　　
益田 紗季子, 北野 翔大, 西端 友香, 外丸 詩野, 石津 明洋
日本病理学会会誌, 111, 1, 214, 214, (一社)日本病理学会, Mar. 2022
Japanese

SLE・抗リン脂質抗体症候群:基礎 全身性エリテマトーデスへのステロイドパルスが好中球細胞外トラップ形成に及ぼす影響　　　　　　　　　　　　　　　
小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 347, 347, (一社)日本リウマチ学会, Mar. 2022
Japanese

ANCA関連血管炎:基礎研究・予後予測因子 ブルトン型チロシンキナーゼ阻害剤チラブルチニブによるMPO-ANCA関連血管炎誘導モデルの発症抑制　　　　　　　　　　　　　　　
中出 一生, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 406, 406, (一社)日本リウマチ学会, Mar. 2022
Japanese

ANCA関連血管炎:基礎研究・予後予測因子 MPO-ANCA関連血管炎モデルにおける新規好中球機能制御化合物薬の抑制効果　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 407, 407, (一社)日本リウマチ学会, Mar. 2022
Japanese

SLE・抗リン脂質抗体症候群:基礎 全身性エリテマトーデスへのステロイドパルスが好中球細胞外トラップ形成に及ぼす影響　　　　　　　　　　　　　　　
小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 347, 347, (一社)日本リウマチ学会, Mar. 2022
Japanese

Possible implication of intermolecular epitope spreading in the production of anti-glomerular basement membrane antibody in anti-neutrophil cytoplasmic antibody-associated vasculitis.
Yuka Nishibata, Mayu Nonokawa, Yuto Tamura, Rio Higashi, Ku Suzuki, Hideyuki Hayashi, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu
Clinical and experimental rheumatology, 40, 4, 691, 704, 04 Feb. 2022, [International Magazine]
English, Scientific journal, OBJECTIVES: Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is sometimes complicated by anti-glomerular basement membrane (GBM) disease. Proteases, including elastase, released from neutrophils activated by ANCA are implicated in the pathogenesis of AAV. Epitopes of anti-GBM antibody exist in the α3-subunit non-collagenous (NC1) domain of collagen type IV [Col (IV)]. This region, called α3(IV)NC1, is structurally cryptic. This study aimed to determine the production mechanism of anti-GBM antibody in AAV. METHODS: We first examined whether α3(IV)NC1 could be revealed by the digestion of formalin-fixed, paraffin-embedded (FFPE) normal kidney sections and Col (IV) by proteases, including neutrophil elastase, using immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA). Next, the reveal of α3(IV)NC1 and the infiltration of CD11c+ macrophages in the affected kidneys were evaluated by IHC and immunofluorescent staining using FFPE sections. Finally, the production of anti-GBM antibody in AAV rats was determined by ELISA. RESULTS: α3(IV)NC1 was revealed by the digestion of FFPE normal kidney sections and Col (IV) by proteases. Although the reveal of α3(IV)NC1 was observed in sclerotic glomeruli regardless of causative diseases, CD11c+ macrophages near α3(IV)NC1 were characteristics of AAV. Anti-GBM antibody was produced subsequent to ANCA in some AAV rats. IHC demonstrated the reveal of α3(IV)NC1 in affected renal tissues and the infiltration of CD11c+ macrophages around the sites. CONCLUSIONS: The collective findings suggest that, in AAV, proteases released from neutrophils activated by ANCA digest Col (IV) and result in the reveal of α3(IV)NC1, CD11c+ macrophages present GBM epitopes, and then the host's immune system produce anti-GBM antibody.

結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の人工知能による鑑別　　　　　　　　　　　　　　　
柏 航, 加藤 千恵次, 西端 友香, 益田 紗季子, 外丸 詩野, 川上 民裕, 石津 明洋
脈管学, 62, 2, 8, 8, (一社)日本脈管学会, Feb. 2022
Japanese

結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の人工知能による鑑別　　　　　　　　　　　　　　　
柏 航, 加藤 千恵次, 西端 友香, 益田 紗季子, 外丸 詩野, 川上 民裕, 石津 明洋
脈管学, 62, 2, 8, 8, (一社)日本脈管学会, Feb. 2022
Japanese

抗ホスファチジルセリン・プロトロンビン複合体抗体による皮膚血管炎動物モデルの完成　　　　　　　　　　　　　　　
川上 民裕, 董 宇鵬, 田村 宥人, 吉成 未来, 西端 友香, 益田 紗季子, 石津 明洋, 外丸 詩野
日本皮膚科学会雑誌, 132, 1, 91, 91, (公社)日本皮膚科学会, Jan. 2022
Japanese

抗ホスファチジルセリン・プロトロンビン複合体抗体による皮膚血管炎動物モデルの完成　　　　　　　　　　　　　　　
川上 民裕, 董 宇鵬, 田村 宥人, 吉成 未来, 西端 友香, 益田 紗季子, 石津 明洋, 外丸 詩野
日本皮膚科学会雑誌, 132, 1, 91, 91, (公社)日本皮膚科学会, Jan. 2022
Japanese

Neutrophil fixation protocols suitable for substrates to detect anti-neutrophil cytoplasmic antibodies by indirect immunofluorescence
Yuka Nishibata, Shun Matsuzawa, Yosuke Satomura, Takeshi Ohtsuka, Motoki Kuhara, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
PATHOLOGY RESEARCH AND PRACTICE, 228, ELSEVIER GMBH, Dec. 2021
English, Scientific journal, ABS T R A C T Anti-neutrophil cytoplasmic antibodies (ANCAs) are autoantibodies that recognize neutrophil cytoplasmic an-tigens. The major ANCA antigens are myeloperoxidase and proteinase 3. Necrotizing small vessel vasculitis accompanied by ANCA production is called ANCA-associated vasculitis (AAV). In addition to AAV, ANCA is sometimes produced in patients with connective tissue diseases, such as systemic lupus erythematosus, and in-flammatory bowel diseases. Indirect immunofluorescence (IIF) and enzyme immunoassay (EIA) have been used to detect ANCAs. Recently, the accuracy of EIA has improved and it has become the gold standard for ANCA detection. However, IIF does not lose its role in ANCA detection because EIA cannot detect ANCAs that recognize antigens other than those coated on the plate. For IIF, neutrophil substrates prepared with two different fixations, namely, ethanol fixation and formalin fixation, are used. There is a recommended protocol for ethanol fixation but not for formalin fixation. This study prepared neutrophil substrates according to the recommended protocol for ethanol fixation and protocols in the literature and original protocols for formalin fixation and then examined ANCA specificity and how storage period would influence the number of cells, antigen distribution, and anti -genicity of the substrates. As a result, the number of cells and antigen distribution did not change after storage for up to 2 months regardless of fixation protocols, whereas a time-dependent decline in ANCA antigenicity and a fixation protocol-dependent difference in ANCA specificity were observed. How neutrophils are fixed on the glass slide needs to be checked upon evaluation of ANCAs by IIF.

Neutrophil fixation protocols suitable for substrates to detect anti-neutrophil cytoplasmic antibodies by indirect immunofluorescence.
Yuka Nishibata, Shun Matsuzawa, Yosuke Satomura, Takeshi Ohtsuka, Motoki Kuhara, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
Pathology, research and practice, 228, 153661, 153661, Dec. 2021, [International Magazine]
English, Scientific journal, Anti-neutrophil cytoplasmic antibodies (ANCAs) are autoantibodies that recognize neutrophil cytoplasmic antigens. The major ANCA antigens are myeloperoxidase and proteinase 3. Necrotizing small vessel vasculitis accompanied by ANCA production is called ANCA-associated vasculitis (AAV). In addition to AAV, ANCA is sometimes produced in patients with connective tissue diseases, such as systemic lupus erythematosus, and inflammatory bowel diseases. Indirect immunofluorescence (IIF) and enzyme immunoassay (EIA) have been used to detect ANCAs. Recently, the accuracy of EIA has improved and it has become the gold standard for ANCA detection. However, IIF does not lose its role in ANCA detection because EIA cannot detect ANCAs that recognize antigens other than those coated on the plate. For IIF, neutrophil substrates prepared with two different fixations, namely, ethanol fixation and formalin fixation, are used. There is a recommended protocol for ethanol fixation but not for formalin fixation. This study prepared neutrophil substrates according to the recommended protocol for ethanol fixation and protocols in the literature and original protocols for formalin fixation and then examined ANCA specificity and how storage period would influence the number of cells, antigen distribution, and antigenicity of the substrates. As a result, the number of cells and antigen distribution did not change after storage for up to 2 months regardless of fixation protocols, whereas a time-dependent decline in ANCA antigenicity and a fixation protocol-dependent difference in ANCA specificity were observed. How neutrophils are fixed on the glass slide needs to be checked upon evaluation of ANCAs by IIF.

免疫疾患の形態病理学的理解 ヒストン皮下注射と抗ホスファチジルセリン・プロトロンビン複合体抗体静脈注射から完成した皮膚血管炎の動物モデル　　　　　　　　　　　　　　　
川上 民裕, 田村 宥人, 董 宇鵬, 吉成 未来, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
日本臨床免疫学会総会プログラム・抄録集, 49回, 58, 58, (一社)日本臨床免疫学会, Oct. 2021
Japanese

膠原病・自己免疫疾患 皮膚血管炎動物モデルの完成と抗ホスファチジルセリン・プロトロンビン複合体抗体
川上 民裕, 田村 宥人, 董 宇鵬, 吉成 未来, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
アレルギー, 70, 6-7, 809, 809, (一社)日本アレルギー学会, Aug. 2021
Japanese

Expression of the immunoproteasome subunit β5i in non-small cell lung carcinomas.
Takayuki Kiuchi, Utano Tomaru, Akihiro Ishizu, Makoto Imagawa, Sari Iwasaki, Akira Suzuki, Noriyuki Otsuka, Yoshihito Ohhara, Ichiro Kinoshita, Yoshihiro Matsuno, Hirotoshi Dosaka-Akita, Masanori Kasahara
Journal of clinical pathology, 74, 5, 300, 306, May 2021, [International Magazine]
English, Scientific journal, AIM: The immunoproteasome is a specific proteasome isoform whose proteolytic activity enhances the generation of antigenic peptides to be presented by major histocompatibility complex class I molecules to CD8+ T cells. Physiologically, it is expressed abundantly in immune cells and is induced in somatic cells by cytokines, especially interferon-γ. Recently, variable expression of immunoproteasomes has been demonstrated in different types of cancers. However, the clinical significance of immunoproteasome expression in malignant tumours is poorly understood. In this study, we performed clinicopathological evaluation of immunoproteasome subunit β5i in non-small cell lung carcinomas (NSCLCs). METHODS: Tumour tissues were collected from 155 patients with NSCLCs, and immunohistochemical analysis for β5i was performed in relation to the prognosis of patients. RESULTS: High expression of β5i was found in about 20% of all NSCLCs and was found significantly more frequently (40%) in the adenocarcinoma subset. High expression of β5i was associated with a better 5-year relative survival rate in patients with pStage I to II adenocarcinoma and was also a significant and independent favourable prognostic factor in adenocarcinoma patients. In addition, when we performed in vitro analysis using NSCLC cell lines, combined treatment with the immunoproteasome-specific inhibitor ONX0914 and the proteasome inhibitor MG132 enhanced cell death in β5i-expressing NSCLC cell lines. CONCLUSION: The expression of immunoproteasome can be explored as both a prognostic factor and a potential therapeutic target in NSCLCs. Since immunoproteasomes have crucial role in the antigen presentation, further studies may help to provide essential knowledge for therapeutic strategies in anticancer immunotherapy.

Anti-phosphatidylserine/prothrombin complex antibodies in patients with cutaneous vasculitis: Possible involvement in the pathogenesis.
Tamihiro Kawakami, Yuto Tamura, Yupeng Dong, Miku Yoshinari, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
The Journal of dermatology, 48, 5, 703, 706, May 2021, [International Magazine]
English, Scientific journal, We assessed the IgG and IgM prevalence of anti-phosphatidylserine/prothrombin complex (aPS/PT) antibodies (Abs) in patients with vasculitis using a novel commercial ELISA kit. To examine whether aPS/PT Abs were involved in the pathogenesis of cutaneous vasculitis, inbred wild-type rats were intravenously administered with a rat IgM class aPS/PT monoclonal Ab established previously or with rat immunoglobulins as controls. To express PS on the surface of vascular endothelium, these rats were given a subcutaneous injection of cell-free histones in advance. Serum IgM aPS/PT Ab levels were elevated in patients with systemic vasculitis with skin involvement and cutaneous arteritis compared to those in patients with systemic vasculitis without skin involvement and healthy controls. There was no significant difference in the serum levels of IgG aPS/PT Abs between the patients and healthy controls. Correspondingly, inbred wild-type rats intravenously administered with the aPS/PT monoclonal IgM Ab after appropriate priming-subcutaneous histone injection developed cutaneous vasculitis. Some rats given rat IgM instead of the aPS/PT monoclonal Ab also developed cutaneous vasculitis, whereas vasculitis did not occur in rats given IgG or only priming by histones. We suggested that IgM aPS/PT Abs could be involved in the pathogenesis of cutaneous vasculitis based on these findings.

RNase in the saliva can affect the detection of severe acute respiratory syndrome coronavirus 2 by real-time one-step polymerase chain reaction using saliva samples.
Yuka Nishibata, Shota Koshimoto, Kenta Ogaki, Erika Ishikawa, Kosuke Wada, Miku Yoshinari, Yuto Tamura, Ryo Uozumi, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
Pathology, research and practice, 220, 153381, 153381, Apr. 2021, [International Magazine]
English, Scientific journal, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a single-stranded RNA virus that causes coronavirus disease 2019, which spread worldwide immediately after the first patient infected with this virus was discovered in Wuhan, China, in December 2019. Currently, polymerase chain reaction (PCR) specimens for the detection of SARS-CoV-2 include saliva, nasopharyngeal swabs, and lower respiratory tract-derived materials such as sputum. Initially, nasopharyngeal swab specimens were applied mainly to the PCR detection of SARS-CoV-2. There was a risk of infection to healthcare workers due to coughing or sneezing by the subjects at the time of sample collection. In contrast, saliva specimens have a low risk of droplet infection and are easy to collect, and their application to PCR testing has been promoted. In this study, we have determined the detection limit of SARS-CoV-2 in saliva samples and examined the effects of storage temperature and storage time of saliva samples on the PCR detection results. As a result, 5 × 103 copies of SARS-CoV-2 could be detected in 1 mL phosphate-buffered saline, whereas 5 × 104 copies of SARS-CoV-2 were needed in 1 mL saliva to detect the virus by real-time one-step PCR. Interestingly, SARS-CoV-2 (5 × 103 copies/mL) could be detected in saliva supplemented with an RNase inhibitor. Concerning the saliva samples supplemented with an RNase inhibitor, the optimal temperature for sample storage was -20 °C, and PCR detection was maintained within 48 h without problems under these conditions. These finding suggest that RNase in the saliva can affect the detection of SARS-CoV-2 by PCR using saliva samples.

ALK転座を伴うdisseminated juvenile xanthogranuloma(D-JXG)の一例　　　　　　　　　　　　　　　
若林 健人, 福井 秀章, 中里 信一, 高桑 恵美, 長 祐子, 本多 昌平, 外丸 詩野, 中澤 温子, 松野 吉宏
日本病理学会会誌, 110, 1, 344, 344, (一社)日本病理学会, Mar. 2021
Japanese

Expression of cathepsins B, D and K in thymic epithelial tumours.
Naoko Yamaguchi, Utano Tomaru, Takayuki Kiuchi, Akihiro Ishizu, Takahiro Deguchi, Noriyuki Otsuka, Satoshi Tanaka, Katsuji Marukawa, Yoshihiro Matsuno, Masanobu Kitagawa, Masanori Kasahara
Journal of clinical pathology, 74, 2, 84, 90, Feb. 2021, [International Magazine]
English, Scientific journal, AIM: Cathepsins are proteases that regulate a wide range of physiological processes, including protein turnover, cell signalling and antigen presentation. Recent studies have shown that cathepsins are highly upregulated in many types of tumours. Of the 15 cathepsins in humans, cathepsins V and S are abundantly expressed in the thymus, and we previously showed that the immunostaining of these cathepsins could serve as diagnostic markers for thymic epithelial tumours. However, little is known about the expression of other cathepsins in thymic epithelial tumours. To determine the diagnostic implications of cathepsins, we performed immunohistochemical analysis of cathepsin B (CTB), cathepsin D (CTD) and cathepsin K (CTK), all of which have been reported to correlate with the progression of squamous cell carcinoma. METHODS: The association between cathepsin expression and clinicopathological features was evaluated in 122 cases of thymoma and thymic carcinoma. RESULTS: CTB and CTD were frequently expressed in type A and type AB thymomas. In contrast, CTB and CTD were significantly less common in type B thymomas than in type A or AB thymomas. In type AB thymomas, the expression of CTB correlated with histological features, and was found predominantly in the type A component. Notably, CTK was expressed most commonly in thymic carcinomas, and patients who died of the disease showed increased expression of CTK. CONCLUSIONS: The expression of CTB and CTD correlated with the histological subtype of thymoma. In addition, the expression of CTK appears to be useful for the diagnosis of thymic carcinomas and as a prognostic marker.

人工知能による結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の鑑別
新海 隼人, 加藤 千恵次, 川上 民裕, 高橋 啓, 西端 友香, 益田 紗季子, 田中 敏, 外丸 詩野, 石津 明洋
脈管学, 61, 1, 1, 2, (一社)日本脈管学会, Jan. 2021
Japanese

人工知能による結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の鑑別
新海 隼人, 加藤 千恵次, 川上 民裕, 高橋 啓, 西端 友香, 益田 紗季子, 田中 敏, 外丸 詩野, 石津 明洋
脈管学, 61, 1, 1, 2, (一社)日本脈管学会, Jan. 2021
Japanese

Decreased Proteasomal Function Induces Neuronal Loss and Memory Impairment.
Utano Tomaru, Tomoki Ito, Yu Ohmura, Kei Higashikawa, Syota Miyajima, Ruka Tomatsu, Tsunehito Higashi, Akihiro Ishizu, Yuji Kuge, Mitsuhiro Yoshioka, Masanori Kasahara
The American journal of pathology, 191, 1, 144, 156, Jan. 2021, [International Magazine]
English, Scientific journal, Alzheimer disease (AD) is a progressive neurodegenerative disorder and the most common type of dementia worldwide. There is considerable evidence of age-related disruption of proteostasis being responsible for the development of AD. The proteasome is a multicatalytic enzyme complex that degrades both normal and damaged proteins, and an age-related decline in its activity has been implicated in age-related pathologies. Although proteasomal dysfunction is assumed to be a key AD hallmark, it remains unclear whether its role in disease onset is causative or secondary. In this study, we demonstrate that mice with proteasomal dysfunction exhibited memory impairment with associated neuronal loss, accumulation of phosphorylated tau, and activation of endoplasmic reticulum (ER) stress-related apoptosis pathways. Impaired proteasomal activity also activated ER stress-related apoptosis pathways in HT-22, a murine hippocampal neuronal cell line. HT-22 cell death, caused by proteasomal inhibition, was prevented by an inhibitor of c-Jun N-terminal kinase, an ER stress-related molecule. Collective evidence suggests that impaired proteasomal activity alters proteostasis, and subsequent ER stress-mediated pathways play pivotal roles in neuronal loss. Because aging decreases proteasomal function, age-related impairment of proteasomes may be involved in the development and progression of AD in elderly patients.

A case of radio-insensitive SMARCA4-deficient thoracic undifferentiated carcinoma with severe right heart failure.
Shotaro Ito, Hajime Asahina, Naoko Yamaguchi, Utano Tomaru, Tadashi Hasegawa, Yutaka Hatanaka, Kanako C Hatanaka, Hiroshi Taguchi, Taisuke Harada, Hiroshi Ohira, Daisuke Ikeda, Hidenori Mizugaki, Eiki Kikuchi, Junko Kikuchi, Jun Sakakibara-Konishi, Naofumi Shinagawa, Satoshi Konno
Respiratory medicine case reports, 32, 101364, 101364, 2021, [International Magazine]
English, SMARCA4-deficient thoracic sarcomatoid tumors were characterized by inactivating mutations of SMARCA4 and often found in the chest of young and middle-aged males with a smoking history. Recently, SMARCA4-deficient thoracic sarcomatoid tumors were reported to represent primarily smoking-associated undifferentiated/de-differentiated carcinomas rather than primary thoracic sarcomas. The main complication of this tumor is compression of the respiratory tract and/or blood vessels. A 39-year-old man presented with a 2-month history of fever and dyspnea. Computed tomography revealed a mediastinal tumor invading the right and left pulmonary arteries. Because of severe right heart failure, we considered him ineligible for bronchoscopy. We scheduled palliative irradiation with 40 Gy/20 Fr to improve hemodynamics and perform endobronchial ultrasound transbronchial needle aspiration later. However, irradiation was ineffective, and his general condition deteriorated quickly and he died after a 7-week hospitalization. An autopsy revealed that the diagnosis was SMARCA4-deficient thoracic undifferentiated carcinoma. It has been reported that this tumor is insensitive to radiotherapy and there were some cases which responded to an immune checkpoint inhibitor. Therefore, when caring for patients with mediastinal tumors that invade and compress the trachea and large vessels, it is important to consider this tumor as a differential diagnosis and try to make a pathological diagnosis as soon as possible.

Association of Neutrophil Extracellular Traps with the Development of Idiopathic Osteonecrosis of the Femoral Head.
Mayu Nonokawa, Tomohiro Shimizu, Miku Yoshinari, Yamato Hashimoto, Yusuke Nakamura, Daisuke Takahashi, Tsuyoshi Asano, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Norimasa Iwasaki, Akihiro Ishizu
The American journal of pathology, 190, 11, 2282, 2289, Nov. 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, Idiopathic osteonecrosis of the femoral head (ONFH) is defined as necrosis of osteocytes due to a non-traumatic ischemia of the femoral head. Iatrogenic glucocorticoid administration and habitual alcohol intake are regarded as risk factors. It has been suggested that glucocorticoid-induced activation of platelets contributes to the local blood flow disturbance of the femoral head. Both activated platelets and alcohol can induce neutrophil extracellular traps (NETs). To determine the association of NETs with the development of idiopathic ONFH, surgically resected femoral heads of patients with idiopathic ONFH and osteoarthritis were assessed for existence of NET-forming neutrophils by immunofluorescence staining. NET-forming neutrophils were present in small vessels surrounding the femoral head of patients with idiopathic ONFH but not osteoarthritis. Moreover, Wistar-Kyoto rats were intravenously injected with NET-forming neutrophils or neutrophils without NET induction, and then the ischemic state of the tissue around the femoral head was evaluated by immunohistochemistry for hypoxia-inducible factor-1α. NET-forming neutrophils circulated into the tissue around the femoral head, and hypoxia-inducible factor-1α expression in the tissue was higher compared with that of rats intravenously administered with neutrophils without NET induction. Furthermore, ischemic change of osteocytes was observed in the femoral head of rats given an i.v. injection of NET-forming neutrophils. The collective findings suggest that NETs are possibly associated with the development of idiopathic ONFH.

特発性大腿骨頭壊死症の発生における好中球細胞外トラップの関与　　　　　　　　　　　　　　　
清水 智弘, 野々川 茉佑, 西端 友香, 益田 紗季子, 高橋 大介, 浅野 毅, 田中 敏, 外丸 詩野, 岩崎 倫政, 石津 明洋
日本整形外科学会雑誌, 94, 8, S1801, S1801, (公社)日本整形外科学会, Sep. 2020
Japanese

中小型血管炎(ANCA関連血管炎) 抗好中球細胞質抗体(ANCA)に続き抗糸球体基底膜(GBM)抗体が産生されるメカニズム　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 64回, 507, 507, (一社)日本リウマチ学会, Aug. 2020
Japanese

中小型血管炎(ANCA関連血管炎) 抗好中球細胞質抗体(ANCA)に続き抗糸球体基底膜(GBM)抗体が産生されるメカニズム　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 64回, 507, 507, (一社)日本リウマチ学会, Aug. 2020
Japanese

Fluvastatin prevents the development of arthritis in env-pX rats via up-regulation of Rho GTPase-activating protein 12.
Shun Tanimura, Mutsumi Nishida, Tatsunori Horie, Tamotsu Kamishima, Hitomi Matsumoto, Yutaka Morimura, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
Experimental and molecular pathology, 115, 104454, 104454, Aug. 2020, [International Magazine]
English, Scientific journal, The pleiotropic effects of statins, including an antiarthritic potential, have been noted. This study aimed to determine the efficacy of statins on rheumatoid arthritis (RA) and clarify how statins affect its pathogenesis. Fluvastatin (500 μg/kg/day) or vehicle was given per os to env-pX rats, which carry the human T-cell leukemia virus type I env-pX gene and spontaneously develop destructive arthritis mimicking RA, for 30 days. Blood sampling and ultrasonography (US) of the ankle joints were conducted on days 0, 10, 20, and 30. On day 30, all rats were euthanized, and the ankle joints were subjected to histological analysis. To clarify how fluvastatin affects the pathogenesis of RA, comprehensive serum exosomal microRNA (miRNA) analysis was performed. Gene expression in the primary culture of synovial fibroblasts derived from arthritic rat and human and non-arthritic rat periarticular tissues was determined quantitatively by real-time reverse transcription-polymerase chain reaction (RT-PCR). As a result, the development of arthritis in env-pX rats was significantly suppressed by fluvastatin, which was evident from the viewpoints of serology, US imaging, and histology. Comprehensive serum exosomal miRNA analysis suggested that the expression of Rho GTPase-activating protein 12 (Arhgap12) was decreased in arthritic env-pX rats but increased with the administration of fluvastatin. Corresponding results were obtained by quantitative RT- PCR using primary culture of synovial fibroblasts. The collective findings suggest that fluvastatin prevents the development of arthritis in env-pX rats via the up-regulation of ARHGAP12. This study suggests that ARHGAP12 can be a possible therapeutic target of RA.

Pharmaceutical immunoglobulins reduce neutrophil extracellular trap formation and ameliorate the development of MPO-ANCA-associated vasculitis.
Ryo Uozumi, Risa Iguchi, Sakiko Masuda, Yuka Nishibata, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
Modern rheumatology, 30, 3, 544, 550, May 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, Objectives: Intravenous immunoglobulin (IVIG) therapy is effective against some autoimmune diseases. We examined the effects of pharmaceutical immunoglobulins on the development of MPO-ANCA-associated vasculitis (MPO-AAV).Methods: Peripheral blood neutrophils were pretreated with 5 mg/ml sulfo-immunoglobulins (IVIG-S) and then exposed to 100 nM phorbol myristate acetate (PMA). Thereafter, neutrophil extracellular traps (NETs) were detected by flow cytometry. Next, Wistar-Kyoto rats were given oral administration of 10 mg/kg/day propylthiouracil for 28 days and intraperitoneal (i.p.) injection of 1 μg PMA on days 0 and 7. These rats were divided into two groups: Group 1 with i.p. injection of 400 mg/kg IVIG-S on days 8-12 and Group 2 with vehicle similarly. ANCA titers were chronologically determined by indirect immunofluorescence. On day 28, all rats were killed to examine NET formation in the peritoneum and the development of AAV.Results: IVIG-S significantly inhibited NET formation induced by PMA in vitro. NET amounts in the peritoneum in Group 1 were significantly smaller than in Group 2, and ANCA titers in Group 1 were significantly lower than in Group 2. The degree of pulmonary hemorrhage in Group 1 was also smaller than in Group 2.Conclusion: IVIG-S reduce NET formation and ameliorate the development of MPO-AAV.

胸腺上皮性腫瘍におけるカテプシンB、D、K発現　　　　　　　　　　　　　　　
山口 直子, 外丸 詩野, 木内 隆之, 石津 明洋, 松野 吉宏, 北川 昌伸, 笠原 正典
日本病理学会会誌, 109, 1, 299, 299, (一社)日本病理学会, Mar. 2020
Japanese

肺基礎疾患を有する肺炎患者における分解抵抗性好中球細胞外トラップの形成　　　　　　　　　　　　　　　
益田 紗季子, 石橋 美郷, 加藤 くるみ, 西端 友香, 田中 敏, 外丸 詩野, 辻野 一三, 石津 明洋
日本病理学会会誌, 109, 1, 308, 308, (一社)日本病理学会, Mar. 2020
Japanese

抗GBM抗体が認識するエピトープの表出に関する検討　　　　　　　　　　　　　　　
西端 友香, 野々川 茉佑, 益田 紗季子, 中沢 大悟, 田中 敏, 外丸 詩野, 石津 明洋
脈管学, 60, 2, 19, 19, (一社)日本脈管学会, Feb. 2020
Japanese

Relationship between lysosomal-associated membrane protein-2 and anti-phosphatidylserine/prothrombin complex antibody in the pathogenesis of cutaneous vasculitis.
Tamihiro Kawakami, Ayaka Kikuchi, Chie Miyabe, Takaharu Ikeda, Sora Takeuchi, Yuto Tamura, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
Clinical and experimental rheumatology, 27 Jan. 2020, [Peer-reviewed], [International Magazine]
English, OBJECTIVES: We investigated the relationship between lysosomal-associated membrane protein-2 (LAMP-2) and anti-phosphatidylserine/prothrombin (PS/PT) antibody in the pathogenesis of cutaneous vasculitis. METHODS: Cell surface LAMP-2 expression of human neutrophils was measured using flow cytometry. Twenty inbred wild-type Wistar-King-Aptekman-Hokudai (WKAH) rats were divided into four groups: Group 1, rabbit IgG injection only as negative control (n=5); Group 2, both histone and rabbit IgG injection (n=5); Group 3, anti-LAMP-2 antibody injection only (n=5); and Group 4, both histone and anti-LAMP-2 antibody injection (n=5). Ten WKAH rats were divided into two groups: Group A, histone, anti-PS/PT antibody, and anti-LAMP-2 antibody injection (n=5), and Group B, histone, anti-PS/PT antibody, and rabbit IgG injection as control (n=5). RESULTS: LAMP-2 expression on human neutrophils was induced by cell-free histone exposure in a dose- and time-dependent manner. Histopathological examination revealed the recruitment of neutrophils in cutaneous small vessels in all Group 4 rats. These observations were not evident in systemic organs other than the skin. LAMP-2 expression on the surface of vascular endothelial cells was evident in Group 2, exclusively in the skin, but not in Group 1. Thrombi were detected in various organs in all Groups A and B rats. However, no apparent thrombi were observed in the skin. CONCLUSIONS: Anti-PS/PT and anti-LAMP-2 antibodies are responsible for independent effector mechanisms in the rats given intravenous injection of cell-free histones. We considered that undetermined factors other than cell-free histones could be required for the induction of cutaneous vasculitis by anti-PS/PT and anti-LAMP-2 antibodies.

A histopathological report of a 16-year-old male with peripheral pulmonary artery stenosis and Moyamoya disease with a homozygous RNF213 mutation.
Kei Takahashi, Junichi Nakamura, Shinya Sakiyama, Toshitaka Nakaya, Takahiro Sato, Taku Watanabe, Hiroshi Ohira, Keishi Makita, Utano Tomaru, Akihiro Ishizu, Ichizo Tsujino
Respiratory medicine case reports, 29, 100977, 100977, 2020, [Peer-reviewed], [International Magazine]
English, Peripheral pulmonary artery stenosis (PPAS) is a rare pulmonary vasculopathy characterized by multiple stenoses and obstructions in the peripheral pulmonary arteries. PPAS often develops in children with congenital diseases such as Williams syndrome and Alagille syndrome; however, recent studies have reported PPAS cases in adults with Moyamoya disease (MMD). Recent genetic studies have demonstrated that ring finger protein 213 (RNF213) is a susceptibility gene for MMD. However, the pathophysiology of combined PPAS and MMD and the relationship between the two diseases remain largely unknown. Here we report a case of PPAS in a 16-year-old male, with a history of MMD, who died suddenly at 24. An autopsy was performed, and remarkable pathological changes were identified in the pulmonary arteries and in other arteries. Furthermore, genetic analysis revealed that the patient had a homozygous c.14576G > A (p.R4859K) mutation in RNF213. This is the first report to demonstrate the histopathology of systemic arteriopathy in a case with MMD and PPAS with a confirmed homozygous RNF213 mutation. We also review immunohistochemical data from the case and discuss how RNF213 mutation could have resulted in the observed vascular abnormalities.

Native myeloperoxidase is required to make the experimental vasculitis model.
Mayu Nonokawa, Ku Suzuki, Hideyuki Hayashi, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu
Arthritis research & therapy, 21, 1, 296, 296, 21 Dec. 2019, [Peer-reviewed], [International Magazine]
English

皮膚血管炎の発症機序における抗リソソーム膜タンパク2抗体(抗LAMP2抗体)と抗ホスファチジルセリン・プロトロンビン複合体抗体(抗PSPT抗体)の役割　　　　　　　　　　　　　　　
川上 民裕, 宮部 千恵, 池田 高治, 菊池 彩花, 西端 友香, 益田 紗季子, 石津 明洋, 中沢 大悟, 外丸 詩野
日本皮膚科学会雑誌, 129, 12, 2533, 2533, (公社)日本皮膚科学会, Nov. 2019
Japanese

Detection of Increased Vascular Signal in Arthritis-Prone Rats Without Joint Swelling Using Superb Microvascular Imaging Ultrasonography.
Tatsunori Horie, Mutsumi Nishida, Shun Tanimura, Tamotsu Kamishima, Erika Tamai, Yutaka Morimura, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
Ultrasound in medicine & biology, 45, 8, 2086, 2093, Aug. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, This study aimed to determine whether ultrasonography (US) can detect increased vascular signal in the synovial tissue before overt synovitis in rheumatoid arthritis (RA). Env-pX rats that spontaneously develop RA-like synovitis were used. Ankle joints of 15 pre-morbid env-pX rats were observed with power Doppler and superb microvascular imaging (SMI) using an ultrahigh-frequency (8-24 MHz) probe. Signal values were counted as the number of pixels. The total number of vessels and vessel area in the synovial tissue were histologically evaluated. Dilated vessels were determined from the mean value of synovial vessels in three wild-type rats. In all env-pX rats, apparent synovial proliferation was not observed. However, vasodilation was evident. Only SMI values were significantly correlated with the number of dilated vessels (r = 0.585, p = 0.022) but not with the total number of vessels. US with SMI using ultrahigh-frequency probe can detect increased vascular signal in the synovial tissue of arthritis-prone rats.

The presence of anti-neutrophil extracellular trap antibody in patients with microscopic polyangiitis.
Fumihiko Hattanda, Daigo Nakazawa, Kanako Watanabe-Kusunoki, Yoshihiro Kusunoki, Haruki Shida, Sakiko Masuda, Saori Nishio, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
Rheumatology (Oxford, England), 58, 7, 1293, 1298, 01 Jul. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, OBJECTIVE: Although ANCA is the major autoantibody in patients with ANCA-associated vasculitis, previous studies have suggested the presence of anti-neutrophil extracellular trap (NET) antibody in patients with microscopic polyangiitis (MPA), one type of ANCA-associated vasculitis. In this study, we aimed to determine the prevalence and pathogenic role of anti-NET antibody (ANETA) in MPA. METHODS: We examined the presence or absence of ANETA in sera obtained from 19 MPA patients by indirect immunofluorescence. We compared the clinical parameters, including age, sex, MPO-ANCA, creatinine, CRP, MPO-DNA complexes and vasculitis activity, in ANETA-positive and ANETA-negative MPA patients. We investigated the serum NET induction and degradation abilities of ANETA-positive and ANETA-negative MPA patients with reference to healthy controls (n = 8). Furthermore, we assessed the relationship between ANETA and the effect of IgG depletion on the serum NET degradation ability. RESULTS: ANETA was present in 10 of the 19 MPA patients. There was no significant difference in the clinical parameters in ANCA-positive and ANCA-negative MPA patients. Although the NET induction ability was higher and the NET degradation ability was lower in MPA sera than those in healthy controls, these abilities were not different between ANETA-positive and ANETA-negative MPA sera. Interestingly, the NET degradation ability in some sera with ANETA was markedly increased by IgG depletion. CONCLUSION: Some MPA patients produce ANETA and some ANETA possess an inhibitory function against the serum NET degradation ability. Although further studies are needed, ANETA is worthy of attention in order to understand the pathophysiology of MPA.

特発性大腿骨頭壊死症の発生における好中球細胞外トラップの関与
野々川 茉佑, 西端 友香, 益田 紗季子, 石津 明洋, 清水 智弘, 高橋 大介, 浅野 毅, 岩崎 倫政, 田中 敏, 外丸 詩野
北海道医学雑誌, 94, 1, 59, 59, 北海道医学会, May 2019
Japanese

プロテアソーム阻害剤使用後の薬剤性心筋症の一剖検例　　　　　　　　　　　　　　　
飯田 圭祐, 土橋 大樹, 木内 隆之, 大塚 紀幸, 石立 尚路, 辻 隆裕, 山口 圭介, 外丸 詩野, 深澤 雄一郎, 笠原 正典
日本病理学会会誌, 108, 1, 454, 454, (一社)日本病理学会, Apr. 2019
Japanese

末梢性T細胞リンパ腫関連血球貪食症候群による劇症肝炎の一例　　　　　　　　　　　　　　　
村山 千咲, 松田 真奈, 大塚 紀幸, 山口 直子, 木内 隆之, 紺野 潤, 田中 敏, 外丸 詩野, 笠原 正典
日本病理学会会誌, 108, 1, 456, 456, (一社)日本病理学会, Apr. 2019
Japanese

血小板増多を示し、腫瘍内に髄外造血巣を伴ったAB型胸腺腫の一例　　　　　　　　　　　　　　　
若林 健人, 福井 秀章, 高桑 恵美, 岡田 宏美, 新垣 雅人, 外丸 詩野, 三橋 智子, 松野 吉宏
日本病理学会会誌, 108, 1, 475, 475, (一社)日本病理学会, Apr. 2019
Japanese

抗GBM抗体が認識するエピトープの表出に関する検討　　　　　　　　　　　　　　　
西端 友香, 東 里緒, 益田 紗季子, 中沢 大悟, 田中 敏, 外丸 詩野, 石津 明洋
日本病理学会会誌, 108, 1, 298, 298, (一社)日本病理学会, Apr. 2019
Japanese

ANCA関連血管炎(AAV)の壊死性病変部における好中球細胞外トラップ(NETs)の存在と病的意義　　　　　　　　　　　　　　　
益田 紗季子, 野々川 茉佑, 西端 友香, 岩崎 沙理, 辻 隆裕, 田中 敏, 外丸 詩野, 川上 民裕, 石津 明洋
日本病理学会会誌, 108, 1, 327, 327, (一社)日本病理学会, Apr. 2019
Japanese

Formation and Disordered Degradation of Neutrophil Extracellular Traps in Necrotizing Lesions of Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis.
Sakiko Masuda, Mayu Nonokawa, Emika Futamata, Yuka Nishibata, Sari Iwasaki, Takahiro Tsuji, Yutaka Hatanaka, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Tamihiro Kawakami, Tatsuya Atsumi, Akihiro Ishizu
The American journal of pathology, 189, 4, 839, 846, Apr. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is characterized by the production of ANCAs and systemic necrotizing vasculitis in small vessels. Disordered regulation of neutrophil extracellular traps (NETs) is critically involved in the pathogenesis of AAV. NETs are web-like DNA decorated with antimicrobial proteins; they are extruded from activated neutrophils. The principal degradation factor of NETs in vivo is DNase I; however, NETs resistant to DNase I can persist in tissues and can lead to the production of ANCAs. Deposition of NETs has been demonstrated in glomerular crescents and necrotizing vasculitis in AAV. Here, the amount of NETs in formalin-fixed, paraffin-embedded tissue sections was examined, and the results for AAV were compared with the results for diseases that should be distinguished from AAV. NETs were more abundant in necrotizing vasculitis of AAV than in non-ANCA-associated vasculitis, or in granulomatous angiitis. Pulmonary granulomas in AAV and non-ANCA-associated diseases were further studied. The amount of NETs was significantly greater in necrotizing granulomas of AAV than in granulomas of sarcoidosis without necrosis. Although NETs were formed in necrotizing granulomas of tuberculosis equivalently to those formed in AAV, they were more susceptible to degradation by DNase I than were NETs in AAV. The formation and disordered degradation of NETs in necrotizing lesions are characteristics of AAV and are possibly related to its pathogenesis.

Epitope recognized by anti-glomerular basement membrane (GBM) antibody in a patient with repeated relapse of anti-GBM disease.
Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Mandkhai Nergui, Xiaoyu Jia, Zhao Cui, Ming-Hui Zhao, Kimimasa Nakabayashi, Akihiro Ishizu
Experimental and molecular pathology, 107, 165, 170, Apr. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, The major epitopes recognized by autoantibodies in anti-glomerular basement membrane (GBM) disease are found in the α3-subunit non-collagenous domain of type IV collagen [α3(IV)NC1], which is present in the glomerular and alveolar basement membranes. These epitopes are structurally cryptic, owing to the hexamer formation of the non-collagenous domain of α3, α4, and α5 subunits and are expressed by the dissociation of the hexamer. Anti-GBM disease usually manifests as a single attack (SA), and we rarely see patients who repeatedly relapse. We recently treated a patient with anti-GBM disease who exhibited repeated relapse (RR). Here, we conducted immunohistochemistry of formalin-fixed paraffin-embedded normal kidney sections and immunoblotting using recombinant human α3(IV)NC1 to compare the epitopes recognized by anti-GBM antibodies in the RR patient and SA patients. Although a clear staining of GBM especially in the connecting basement membrane of Bowman's capsule was observed when IgGs of SA patients were used as primary antibodies, such staining was not obtained when IgG of the RR patient was employed. In immunoblotting of α3(IV)NC1 using the IgG of the RR patient as a primary antibody, an 18-kDa band was detected besides the 56.8-kDa band corresponding to the whole-size α3(IV)NC1. Whereas the 56.8-kDa band disappeared after digestion of the recombinant α3(IV)NC1 by protease, the 18-kDa band remained. Furthermore, the 18-kDa band was not detected by a commercially available anti-α3(IV)NC1 monoclonal antibody. These findings suggest that the IgG of the RR patient recognizes the epitope distinct from that recognized by the anti-α3(IV)NC1 monoclonal antibody.

抗糸球体基底膜抗体病(抗GBM病)の頻回再発症例の抗GBMが認識するエピトープ　　　　　　　　　　　　　　　
西端 友香, 東 里緒, 益田 紗季子, 中沢 大悟, 田中 敏, 外丸 詩野, 中林 公正, 石津 明洋
脈管学, 59, 3, 19, 19, (一社)日本脈管学会, Mar. 2019
Japanese

ヒストンは好中球細胞上にLAMP2を表出し、抗LAMP2抗体と連携して皮膚小血管炎の発症機序に関与している　　　　　　　　　　　　　　　
川上 民裕, 竹内 そら, 菊池 彩翔, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋
脈管学, 59, 3, 22, 22, (一社)日本脈管学会, Mar. 2019
Japanese

ANCA関連血管炎の壊死性病変部における好中球細胞外トラップの存在と病的意義　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 中沢 大悟, 外丸 詩野, 川上 民裕, 渥美 達也, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 63回, 777, 777, (一社)日本リウマチ学会, Mar. 2019
Japanese

Author Correction: Pathogenesis and therapeutic interventions for ANCA-associated vasculitis.
Daigo Nakazawa, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
Nature reviews. Rheumatology, 15, 2, 123, 123, Feb. 2019, [International Magazine]
English, In the originally published online version of this article there were errors in the Supplementary Information. All three Supplementary Tables had incorrectly numbered references. These errors have now been corrected in the HTML and PDF versions of the manuscript.

The Diagnostic and Clinical Utility of the Myeloperoxidase-DNA Complex as a Biomarker in Otitis Media With Antineutrophil Cytoplasmic Antibody-associated Vasculitis.
Shinya Morita, Yuji Nakamaru, Daigo Nakazawa, Masanobu Suzuki, Kimiko Hoshino, Atsushi Fukuda, Fumihiko Hattanda, Kanako Kusunoki, Utano Tomaru, Akihiro Ishizu, Akihiro Homma
Otology & neurotology : official publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology, 40, 2, e99-e106, e106, Feb. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, OBJECTIVE: This prospective study aimed to evaluate the diagnostic and clinical utility of the myeloperoxidase (MPO)-DNA complex as a NETosis-derived product in the middle ear fluid of patients with otitis media with antineutrophil cytoplasmic antibody-associated vasculitis (OMAAV). STUDY DESIGN: Prospective study. SETTING: Tertiary referral center. PATIENTS: Twenty-two patients diagnosed with OMAAV. INTERVENTION: Collection of the fluid samples from middle ear. MAIN OUTCOME MEASURE: The levels of the MPO-DNA complex in the fluid samples were quantified using an enzyme-linked immunosorbent assay. RESULTS: Patients with both systemic and localized forms of OMAAV showed significantly higher levels of the MPO-DNA complex compared to the controls (p < 0.001 and p = 0.002, respectively). In particular, they showed significantly higher levels of MPO-DNA complex compared to the controls, regardless of serum antineutrophil cytoplasmic antibody status (p < 0.001 and p < 0.001, respectively) or immunosuppressive therapy (p < 0.001 and p < 0.001, respectively) at the time of sampling. An optical density cutoff value of 0.16 at 405 nm according to the receiver operating characteristic curve showed a sensitivity of 86.4%, specificity of 95.5%, positive predictive value of 95.0% and negative predictive value of 87.5% for the diagnosis of OMAAV. Significant positive correlations were observed between the levels of MPO-DNA complex and the values for air conduction - (r = 0.49, p = 0.022) and bone conduction - pure tone average thresholds (r = 0.45, p = 0.035). CONCLUSIONS: The detection and quantification of the MPO-DNA complex in the otitis media fluid may aid in providing a definite diagnosis as well as predicting the activity and severity of OMAAV.

Pathogenesis and therapeutic interventions for ANCA-associated vasculitis.
Daigo Nakazawa, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
Nature reviews. Rheumatology, 15, 2, 91, 101, Feb. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) affects systemic small vessels and is accompanied by the presence of ANCAs in the serum. This disease entity includes microscopic polyangiitis, granulomatosis with polyangiitis, eosinophilic granulomatosis with polyangiitis and drug-induced AAV. Similar to other autoimmune diseases, AAV develops in patients with a predisposing genetic background who have been exposed to causative environmental factors. The mechanism by which ANCAs cause vasculitis involves ANCA-mediated excessive activation of neutrophils that subsequently release inflammatory cytokines, reactive oxygen species and lytic enzymes. In addition, this excessive activation of neutrophils by ANCAs induces formation of neutrophil extracellular traps (NETs). Although NETs are essential elements in innate immunity, excessive NET formation is harmful to small vessels. Moreover, NETs are involved not only in ANCA-mediated vascular injury but also in the production of ANCAs themselves. Therefore, a vicious cycle of NET formation and ANCA production is considered to be involved in the pathogenesis of AAV. In addition to this role of NETs in AAV, some other important discoveries have been made in the past few years. Incorporating these new insights into our understanding of the pathogenesis of AAV is needed to fully understand and ultimately overcome this disease.

Restricted Expression of the Thymoproteasome Is Required for Thymic Selection and Peripheral Homeostasis of CD8+ T Cells.
Utano Tomaru, Saori Konno, Syota Miyajima, Rikuto Kimoto, Mari Onodera, Shizuka Kiuchi, Shigeo Murata, Akihiro Ishizu, Masanori Kasahara
Cell reports, 26, 3, 639, 651, 15 Jan. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, The thymoproteasome subunit β5t is specifically expressed in cortical thymic epithelial cells (TECs) and generates unique peptides to support positive selection. In this study, using a mouse model ubiquitously expressing β5t, we showed that aberrant expression of self-peptides generated by β5t affects CD8+ T cell homeostasis, including thymic selection and maintenance of the peripheral naive pool of CD8+ T cells. In mice in which β5t was expressed both in cortical and medullary TECs, the abundance of CD8+ lineage thymocytes was reduced, and extra-thymic expression of β5t caused accumulation of CD8+ T cells with the memory or exhausted phenotype and induced autoreactive T cell responses. We found that thymoproteasomes are essential for positive selection but that the subsequent change in peptide repertoire in the medulla is also crucial for thymic selection and that β5t-derived peptide must be confined to the thymus to avoid autoimmunity in peripheral tissues.

Detection of Autoreactive Type II NKT Cells: A Pilot Study of Comparison Between Healthy Individuals and Patients with Vasculitis.
Yusuke Nishioka, Takaomi Sonoda, Haruki Shida, Yoshihiro Kusunoki, Fumihiko Hattanda, Shun Tanimura, Ryo Uozumi, Mai Yamada, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 93, 11, 1157, 1164, Nov. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, NKT cells are defined as T cells that recognize hydrophobic antigens presented by class I MHC-like molecules, including CD1d. Among CD1d-restricted NKT cells, type I and type II subsets have been noted. CD1d-restricted type I NKT cells are regarded as pro-inflammatory cells in general. On the contrary, accumulated evidence has demonstrated an anti-inflammatory property of CD1d-restricted type II NKT cells. In our earlier study using a rat model with vasculitis, we demonstrated the pro-inflammatory function of CD1d-restricted type II NKT cells and identified that one such cell recognized P518-532 of rat sterol carrier protein 2 (rSCP2518-532 ), which appeared on vascular endothelial cells presented by CD1d. Based on this evidence, we attempted to detect human CD1d-restricted type II NKT cells in peripheral blood using hSCP2518-532 , the human counterpart of rSCP2518-532, together with a CD1d tetramer in flow cytometry. First, we determined the binding of hSCP2518-532 to CD1d. Next, we detected CD3-positive hSCP2518-532 -loaded CD1d (hSCP2518-532 /CD1d) tetramer-binding cells in peripheral blood of healthy donors. The abundance of TGF-β-producing cells rather than TNF-α-producing cells in CD3-positive hSCP2518-532 /CD1d tetramer-binding cells suggests the anti-inflammatory property of SCP2-loaded CD1d (SCP2/CD1d) tetramer-binding type II NKT cells in healthy individuals. Furthermore, we compared cytokine profile between healthy individuals and patients with vasculitis in a pilot study. Interestingly, the percentage of TGF-β-producing cells in SCP2/CD1d tetramer-binding type II NKT cells in vasculitic patients was significantly lower than that in healthy controls despite the greater number of these cells. Although further studies to clarify the mechanism and significance of this phenomenon are needed, SCP2/CD1d tetramer-binding type II NKT cells in peripheral blood should be examined in more detail to understand the pathophysiology of vasculitides in humans. © 2018 International Society for Advancement of Cytometry.

Clinicopathological significance of PSF3 expression in uterine endometrial carcinomas.
Park JK, Otsuka N, Tomaru U, Suzuki H, Azuma M, Okamoto K, Yamashiro K, Kasahara M
Human pathology, 80, 104, 112, Oct. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, PSF3 (Partner of SLD Five 3) is a member of the heterotetrameric complex termed GINS. Previous studies have shown that PSF3 is up-regulated in several cancers and is associated with tumor malignancy. However, the clinicopathological significance of PSF3 expression in endometrial lesions is still poorly understood. To investigate whether PSF3 could serve as a useful biomarker for endometrial carcinomas, we performed immunohistochemical analysis of PSF3 expression. In 155 cases of endometrial carcinomas (ECs), the mean tumor proportion score of PSF3 expression was 30.7% in G1 endometrioid carcinoma, 55.0% in G2 endometrioid carcinoma, 59.0% in G3 endometrioid carcinoma, and 58.9% in nonendometrioid carcinomas. In 25 cases of atypical hyperplasia, the mean tumor proportion score of PSF3 expression was significantly lower (10.4%). High expression of PSF3 was associated with more advanced pathologic T stage (P = .000), lymphatic invasion (P = .001), and poor clinical outcomes such as shorter relapse-free survival (P = .000) and overall survival (P = .001). When we compared the immunostaining of PSF3 and Ki-67, the proportions of PSF3-positive cells in tumor epithelial cells were comparable to those of Ki-67-positive cells. However, PSF3-positive cells were selectively found in tumor cells, whereas Ki-67-positive cells were also found in tumor stromal cells. These results demonstrated that PSF3 immunostaining was valuable as a histopathologic marker for differential diagnosis between atypical hyperplasia and ECs, for tumor histologic grading, and for determining a patient's prognosis. PSF3 may play a crucial role in tumor progression in EC.

Anti-neutrophil extracellular trap antibody in a patient with relapse of anti-neutrophil cytoplasmic antibody-associated vasculitis: A case report
Haruki Shida, Nobuhiro Hashimoto, Yoshihiro Kusunoki, Fumihiko Hattanda, Yayoi Ogawa, Terumasa Hayashi, Daigo Nakazawa, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
BMC Nephrology, 19, 1, 145, BioMed Central Ltd., 22 Jun. 2018, [Peer-reviewed]
English, Scientific journal, Background: Neutrophil extracellular traps (NETs) are web-like DNA decorated with antimicrobial proteins, such as myeloperoxidase (MPO), which are extruded from activated neutrophils. Although NETs are essential in innate immunity, an excessive formation of NETs has adverse effects, e.g., induction of anti-neutrophil cytoplasmic antibody (ANCA), to the hosts. Since ANCA can induce NET formation in the primed neutrophils, a positive feedback loop can be formed between NETs and ANCA, which is called "ANCA-NETs vicious cycle." Case presentation: A 79-year-old Japanese woman developed hydralazine-induced pauci-immune necrotizing crescentic glomerulonephritis with MPO-ANCA. Although the illness improved after cessation of hydralazine, MPO-ANCA-associated vasculitis relapsed 16 months later. Remission was achieved 5 months after beginning of administration of prednisone. In order to determine the involvement of ANCA-NETs vicious cycle in this patient, we examined NET degradation and induction activities in sera obtained at the disease onset (Serum A
MPO-ANCA, 107 IU/ml), at relapse (Serum B
MPO-ANCA, 195 IU/ml), at 3 months after treatment (Serum C
MPO-ANCA, 4.5 IU/ml), and at remission (Serum D
MPO-ANCA, 2.4 IU/ml). NET degradation activity was low in the all sera. NET induction activity was high in Sera A, B, and C but not in D. Additionally, we demonstrated the presence of anti-NET antibody (ANETA) in Sera B and C but not in A or D. Conclusions: The collective findings suggest NET induction potential of ANETA in the present patient and that the ANETA could contribute to the enhancement of NETs resulting in amplification of the ANCA-NETs vicious cycle.

Pleuloparenchymal fibroelastosisの病理解剖5症例　　　　　　　　　　　　　　　
土橋 大樹, 木内 隆之, 桑原 健, 牧田 啓史, 仲川 心平, 鈴木 雅, 大塚 紀幸, 田中 敏, 外丸 詩野, 笠原 正典
日本病理学会会誌, 107, 1, 528, 528, (一社)日本病理学会, Apr. 2018
Japanese

致死的な腹腔内出血を来した顕微鏡的多発血管炎の一剖検例　　　　　　　　　　　　　　　
伊丹 久実, 木内 隆之, 外丸 詩野, 大塚 紀幸, 田中 敏, 石津 明洋, 笠原 正典
日本病理学会会誌, 107, 1, 535, 535, (一社)日本病理学会, Apr. 2018
Japanese

ANCA関連血管炎(AAV)の壊死性病変部における好中球細胞外トラップ(NETs)の存在と病的意義　　　　　　　　　　　　　　　
益田 紗季子, 野々川 茉佑, 西端 友香, 岩崎 沙理, 辻 隆裕, 田中 敏, 外丸 詩野, 川上 民裕, 石津 明洋
日本病理学会会誌, 107, 1, 330, 330, (一社)日本病理学会, Apr. 2018
Japanese

免疫グロブリン大量静注療法(IVIG)は好中球細胞外トラップ(NETs)の抑制によりMPO-ANCA関連血管炎を抑制する　　　　　　　　　　　　　　　
魚住 諒, 井口 理彩, 益田 紗季子, 西端 友香, 谷村 瞬, 中沢 大悟, 外丸 詩野, 石津 明洋
日本病理学会会誌, 107, 1, 331, 331, (一社)日本病理学会, Apr. 2018
Japanese

Elevated Level of Myeloperoxidase-Deoxyribonucleic Acid Complex in the Middle Ear Fluid Obtained from Patients with Otitis Media Associated with Antineutrophil Cytoplasmic Antibody-associated Vasculitis
Shinya Morita, Yuji Nakamaru, Daigo Nakazawa, Fumihiko Hattanda, Haruki Shida, Yoshihiro Kusunoki, Kanako Watanabe, Sakiko Masuda, Dai Takagi, Masanobu Suzuki, Kimiko Hoshino, Atsushi Fukuda, Utano Tomaru, Akihiro Homma, Akihiro Ishizu
Otology and Neurotology, 39, 4, e257, e262, Lippincott Williams and Wilkins, 01 Apr. 2018, [Peer-reviewed]
English, Scientific journal, Objective:The purpose was to explore the presence of myeloperoxidase (MPO)-deoxyribonucleic acid (DNA) complex as a surrogate marker of neutrophil extracellular traps (NETs) in the middle ear fluid, and to clarify the correlation between its quantifiable level and hearing outcome in patients with otitis media associated with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV).Study Design:Prospective study.Setting:Tertiary referral center.Patients:Nine AAV patients presenting with otitis media.Intervention:Collection of the fluid samples from middle ear.Main Outcome Measure:The quantifiable levels of MPO-DNA complex using an enzyme-linked immunosorbent assay.Results:The quantifiable levels of MPO-DNA complex in patients with AAV were significantly higher than those in controls (p <
0.001). In particular, both ANCA-positive and-negative cases indicated higher levels of MPO-DNA complex compared with the controls (p = 0.004 and p = 0.006, respectively). The significant negative correlations were observed between the level of MPO-DNA complex and the functional hearing values for air (r =-0.82, p = 0.009) and bone conduction (r =-0.73, p = 0.028), respectively.Conclusion:This analysis is the first to reveal the presence of elevated levels of MPO-DNA complex in the middle ear fluid, suggesting the pathogenic role of NETs in otitis media associated with AAV. NETs may be a valuable biomarker for use in clinical decision-making and predicting hearing outcome, regardless of ANCA status.

Brain-derived neurotrophic factor induces angiogenin secretion and nuclear translocation in human umbilical vein endothelial cells
Ayako Mori, Yusuke Nishioka, Mai Yamada, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Naoyuki Honma, Takanori Moriyama, Akihiro Ishizu
Pathology Research and Practice, 214, 4, 521, 526, Elsevier GmbH, 01 Apr. 2018, [Peer-reviewed]
English, Scientific journal, Brain-derived neurotrophic factor (BDNF) is a well-known humoral protein that induces growth of neurons. Recent studies have suggested that BDNF could act as an angiogenesis inducer similar to vascular endothelial growth factor (VEGF). Angiogenin is a strong mediator of angiogenesis. It has particular characteristics both as a secreted protein and a transcription factor. After being incorporated into the cytoplasm, angiogenin is immediately transferred to the nucleus and then mediates the angiogenic effects of angiogenesis inducers, including VEGF. The aim of this study is to determine the association between BDNF and angiogenin. At first, we determined the secretion of angiogenin from human umbilical vein endothelial cells (HUVEC) induced by BDNF with enzyme-linked immunosorbent assay. Next, we determined BDNF-induced nuclear translocation of angiogenin by immunofluorescent staining. In addition, we examined the mRNA expression of angiogenin in HUVEC before and after BDNF stimulation by quantitative reverse transcriptase-polymerase chain reaction. As a result, we noted that BDNF induced angiogenin secretion and nuclear translocation without an increase in the mRNA expression in HUVEC. Furthermore, we demonstrated that BDNF-induced HUVEC proliferation was significantly suppressed when neomycin, a specific inhibitor of nuclear translocation of angiogenin, was administered. These findings indicate that nuclear translocation of angiogenin is critically involved in BDNF-induced proliferation of HUVEC. In conclusion, angiogenin contributes to angiogenesis induced by BDNF.

CD1d-restricted type II NKT cells reactive with endogenous hydrophobic peptides
Yusuke Nishioka, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
Frontiers in Immunology, 9, 548, Frontiers Media S.A., 15 Mar. 2018, [Peer-reviewed]
English, NKT cells belong to a distinct subset of T cells that recognize hydrophobic antigens presented by major histocompatibility complex class I-like molecules, such as CD1d. Because NKT cells stimulated by antigens can activate or suppress other immunocompetent cells through an immediate production of a large amount of cytokines, they are regarded as immunological modulators. CD1d-restricted NKT cells are classified into two subsets, namely, type I and type II. CD1d-restricted type I NKT cells express invariant T cell receptors (TCRs) and react with lipid antigens, including the marine sponge-derived glycolipid a-galactosylceramide. On the contrary, CD1d-restricted type II NKT cells recognize a wide variety of antigens, including glycolipids, phospholipids, and hydrophobic peptides, by their diverse TCRs. In this review, we focus particularly on CD1d-restricted type II NKT cells that recognize endogenous hydrophobic peptides presented by CD1d. Previous studies have demonstrated that CD1d-restricted type I NKT cells usually act as pro-inflammatory cells but sometimes behave as anti-inflammatory cells. It has been also demonstrated that CD1d-restricted type II NKT cells play opposite roles to CD1d-restricted type I NKT cells
thus, they function as anti-inflammatory or pro-inflammatory cells depending on the situation. In line with this, CD1d-restricted type II NKT cells that recognize type II collagen peptide have been demonstrated to act as anti-inflammatory cells in diverse inflammation-induction models in mice, whereas pro-inflammatory CD1d-restricted type II NKT cells reactive with sterol carrier protein 2 peptide have been demonstrated to be involved in the development of small vessel vasculitis in rats.

ANCA関連血管炎の壊死性病変部におけるNETsの存在と病的意義　　　　　　　　　　　　　　　
益田 紗季子, 野々川 茉佑, 西端 友香, 岩崎 沙理, 辻 隆裕, 田中 敏, 外丸 詩野, 川上 民裕, 石津 明洋
脈管学, 58, 3, 35, 35, (一社)日本脈管学会, Mar. 2018
Japanese

プロテアーゼによる抗糸球体基底膜抗体病の隔絶抗原の表出　　　　　　　　　　　　　　　
西端 友香, 植松 千浩, 益田 紗季子, 田中 敏, 外丸 詩野, 石津 明洋
脈管学, 58, 3, 36, 37, (一社)日本脈管学会, Mar. 2018
Japanese

血管炎1:血管炎のバイオマーカー・病態解析 免疫グロブリン製剤は好中球細胞外トラップ(NETs)の形成抑制を介してMPO-ANCA関連血管炎(MPO-AAV)の発症を抑制する　　　　　　　　　　　　　　　
魚住 諒, 益田 紗季子, 西端 友香, 谷村 瞬, 中沢 大悟, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 62回, 433, 433, (一社)日本リウマチ学会, Mar. 2018
Japanese

リウマチ性疾患の画像2:関節エコーその他 超高周波プローブを用いたSuperb Micro-vascular Imagingによる超音波検査でのラット足関節の早期関節炎診断　　　　　　　　　　　　　　　
西田 睦, 谷村 瞬, 神島 保, 西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 渥美 達也, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 62回, 485, 485, (一社)日本リウマチ学会, Mar. 2018
Japanese

スタチンの関節炎抑制効果とその機序の解明　　　　　　　　　　　　　　　
谷村 瞬, 西田 睦, 神島 保, 西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 渥美 達也, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 62回, 778, 778, (一社)日本リウマチ学会, Mar. 2018
Japanese

Vanishing Immunoglobulins: The Formation of Pauci-Immune Lesions in Myeloperoxidase-Antineutrophil Cytoplasmic Antibody-Associated Vasculitis
Emika Futamata, Sakiko Masuda, Yuka Nishibata, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu
Nephron, 138, 4, 328, 330, S. Karger AG, 01 Mar. 2018, [Peer-reviewed]
English

Superb micro-vascular imaging法のシグナル強度と関節内血管径の関係性 関節リウマチの実験動物モデル(Relationship between Superb Micro-vascular Imaging Signal Quantity and Vessel Diameter in the Joints: Experimental Animal Models of Rheumatoid Arthritis)　　　　　　　　　　　　　　　
玉井 絵里香, 神島 保, 森村 豊, 堀江 達則, 谷村 瞬, 西田 睦, 石津 明洋, 外丸 詩野
日本放射線技術学会雑誌, 73, 9, 855, 855, (公社)日本放射線技術学会, Sep. 2017
English

Measurement of NET formation in vitro and in vivo by flow cytometry
Sakiko Masuda, Sakika Shimizu, Junji Matsuo, Yuka Nishibata, Yoshihiro Kusunoki, Fumihiko Hattanda, Haruki Shida, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
CYTOMETRY PART A, 91A, 8, 822, 829, WILEY, Aug. 2017, [Peer-reviewed]
English, Scientific journal, Neutrophil extracellular traps (NETs) are extracellular chromatin fibers adorned with antimicrobial proteins, such as myeloperoxidase (MPO), which are extruded from activated neutrophils. NETosis is the metamorphosis of neutrophils with NET formation that follows decondensation of DNA and rupture of the plasma membrane. Although NETs play important roles in innate immunity, excessive formation of NETs can be harmful to the hosts. Until now, various methods for evaluation of NETs have been reported. Although each has a virtue, the gold standard has not been established. Here we demonstrate a simple, objective, and quantitative method to detect NETs using flow cytometry. This method uses a plasma membrane-impermeable DNA-binding dye, SYTOX Green. SYTOX Green-positive cells were detected in human peripheral polymorphonuclear cells exposed to a NET inducer, phorbol 12-myristate 13-acetate (PMA). The number of SYTOX Green-positive cells was increased depending on the exposure duration and concentrations of PMA. Furthermore, co-localization of MPO and plasma membrane-appendant DNA of SYTOX Green-positive cells was demonstrated. Moreover, a NET inhibitor, diphenylene iodonium, could significantly reduce the number of SYTOX Green-positive cells induced by PMA. The collective evidence suggests that SYTOX Green-positive cells include neutrophils that formed NETs. The established method could detect neutrophils that underwent NETosis but not early apoptosis with equivalence in quantification to another well-used image analysis, which is based on fluorescent staining. Additionally, NETs that were formed in vivo were also detectable by this method. It is conceivable that the established method will bring us better understanding of the relation between NETosis and human diseases. (c) 2017 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC.

Establishment of a rat model of thrombosis induced by intravenous injection of anti-phosphatidylserine-prothrombin complex antibody
Mai Yamada, Tamihiro Kawakami, Kohei Takashima, Yusuke Nishioka, Yuka Nishibata, Sakiko Masuda, Shigeru Yoshida, Utano Tomaru, Akihiro Ishizu
RHEUMATOLOGY, 56, 6, 1013, 1018, OXFORD UNIV PRESS, Jun. 2017, [Peer-reviewed]
English, Scientific journal, Objective. Recent studies have suggested that aPS-PT antibody is one of the most relevant autoantibodies to APS. This study aimed to demonstrate the pathogenicity of aPS-PT antibody in vivo.
Methods. At first, cultured rat vascular endothelial cells (RECs) were exposed to calf thymus-derived histones. Two hours later, lactate dehydrogenase release from the RECs and expression of PS on the cell surface were assessed. Next, we administered an i.v. injection of calf thymus-derived histones into Wistar rats (12.5 mu g/g weight of 8-week-old female rats), and 2 h later they were given an i.v. injection of aPS-PT mAb (1.25 mg/g weight, n = 6) or an equal dose of rat IgM as controls (n = 5). Three days later, histological examination was conducted.
Results. Calf thymus-derived histones (> 12.5 mg/ml) could injure RECs in vitro. Simultaneously, annexin V could bind to the RECs; thereby, this result indicated that cell-free histone exposure of vascular endothelial cells induced cell surface expression of PS, which is naturally present inside the plasma membrane. Thrombosis developed with higher frequency in the rats given an i. v. injection of aPS-PT mAb than in controls.
Conclusion. We established a rat model of thrombosis induced by i. v. injection of aPS-PT mAb.

Prediction of response to remission induction therapy by gene expression profiling of peripheral blood in Japanese patients with microscopic polyangiitis
Akihiro Ishizu, Utano Tomaru, Sakiko Masuda, Ken-ei Sada, Koichi Amano, Masayoshi Harigai, Yasushi Kawaguchi, Yoshihiro Arimura, Kunihiro Yamagata, Shoichi Ozaki, Hiroaki Dobashi, Sakae Homma, Yasunori Okada, Hitoshi Sugiyama, Joichi Usui, Naotake Tsuboi, Seiichi Matsuo, Hirofumi Makino
ARTHRITIS RESEARCH & THERAPY, 19, 1, 117, BIOMED CENTRAL LTD, May 2017, [Peer-reviewed]
English, Scientific journal, Background: Microscopic polyangiitis (MPA), which is classified as an anti-neutrophil cytoplasmic antibody (ANCA)associated small vessel vasculitis, is one of the most frequent primary vasculitides in Japan. We earlier nominated 16 genes (IRF7, IFIT1, IFIT5, OASL, CLC, GBP-1, PSMB9, HERC5, CCR1, CD36, MS4A4A, BIRC4BP, PLSCR1, DEFA1/DEFA3, DEFA4, and COL9A2) as predictors of response to remission induction therapy against MPA. The aim of this study is to determine the accuracy of prediction using these 16 predictors.
Methods: Thirty-nine MPA patients were selected randomly and retrospectively from the Japanese nationwide RemIT-JAV-RPGN cohort and enrolled in this study. Remission induction therapy was conducted according to the Guidelines of Treatment for ANCA-Associated Vasculitis published by the Ministry of Health, Labour, and Welfare of Japan. Response to remission induction therapy was predicted by profiling the altered expressions of the 16 predictors between the period before and 1 week after the beginning of treatment. Remission is defined as the absence of clinical manifestations of active vasculitis (Birmingham Vasculitis Activity Score 2003: 0 or 1 point). Persistent remission for 18 months is regarded as a "good response," whereas no remission or relapse after remission is regarded as a "poor response."
Results: "Poor" and "good" responses were predicted in 7 and 32 patients, respectively. Five out of 7 patients with "poor" prediction and 1 out of 32 patients with "good" prediction experienced relapse after remission. One out of 7 patients with "poor" prediction was not conducted to remission. Accordingly, the sensitivity and specificity to predict poor response was 85.7% (6/7) and 96.9% (31/32), respectively.
Conclusions: Response to remission induction therapy can be predicted by monitoring the altered expressions of the 16 predictors in the peripheral blood at an early point of treatment in MPA patients.

比較的急速に進行した脱髄疾患の1剖検例　　　　　　　　　　　　　　　
松林 里佳, 小林 珠代, 木内 隆之, 外丸 詩野, 柳内 充, 大塚 紀幸, 田中 敏, 辻 隆裕, 深澤 雄一郎, 笠原 正典
日本病理学会会誌, 106, 1, 508, 508, (一社)日本病理学会, Mar. 2017
Japanese

フローサイトメトリーによる定量的好中球細胞外トラップ測定法　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 松尾 淳司, 外丸 詩野, 石津 明洋
日本病理学会会誌, 106, 1, 350, 350, (一社)日本病理学会, Mar. 2017
Japanese

血管炎の実験動物モデル 抗PSPT抗体は、正常ラットに血栓を発症させる　　　　　　　　　　　　　　　
川上 民裕, 山田 真衣, 高島 滉平, 西岡 佑介, 西端 友香, 益田 紗季子, 吉田 繁, 外丸 詩野, 石津 明洋
脈管学, 57, 2, 22, 22, (一社)日本脈管学会, Feb. 2017
Japanese

Expression of cathepsins V and S in thymic epithelial tumors
Shizuka Kiuchi, Utano Tomaru, Akihiro Ishizu, Makoto Imagawa, Takayuki Kiuchi, Sari Iwasaki, Akira Suzuki, Noriyuki Otsuka, Takahiro Deguchi, Tomohiro Shimizu, Katsuji Marukawa, Yoshihiro Matsuno, Masanori Kasahara
HUMAN PATHOLOGY, 60, 66, 74, W B SAUNDERS CO-ELSEVIER INC, Feb. 2017, [Peer-reviewed]
English, Scientific journal, Cathepsins are a group of proteolytic enzymes of the endosomal/lysosomal pathway involved in the thymic development of T cells restricted by major histocompatibility complex class II molecules. In the normal thymus, cathepsin V (CTV) and cathepsin S (CTS) are expressed in cortical and medullary epithelial cells, respectively. To investigate whether cathepsins could serve as a diagnostic marker, we performed immunohistochemical analysis for CTV and CTS in 77 cases of thymic epithelial tumors. Almost all cases (59/ 60) of thymoma expressed CTV, whereas 28 of 60 cases of thymoma expressed CTS. Notably, CTS was expressed in most cases of type A and type AB thymomas, but not in type B thymoma. The expression of cathepsins in type AB thymoma showed a clear correlation with histologic features; CTV was found predominantly in the type B component, and CTS was frequently expressed in the type A component. In thymic carcinoma, CTV was expressed in less than half cases (7/17), and the ratio of CTS-positive cases was equivalent to that of thymoma (8/17). Cases of CTV-negative thymic carcinoma tended to have a higher incidence of recurrence than did CTV-positive cases. Although further studies with a larger number of cases are required to confirm the utility of cathepsin immunostaining, CTV and CTS appear to serve as auxiliary diagnostic and/or prognostic markers in thymic epithelial tumors. (C) 2016 Elsevier Inc. All rights reserved.

Type II Natural Killer T Cells that Recognize Sterol Carrier Protein 2 Are Implicated in Vascular Inflammation in the Rat Model of Systemic Connective Tissue Diseases
Yusuke Nishioka, Madoka Yamaguchi, Ai Kawakami, Maya Munehiro, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
AMERICAN JOURNAL OF PATHOLOGY, 187, 1, 176, 186, ELSEVIER SCIENCE INC, Jan. 2017, [Peer-reviewed]
English, Scientific journal, We previously generated a rat model that developed systemic connective tissue diseases, including synovitis, myositis, and small-vessel vasculitis (SVV), and established a vascular endothelial cell-reactive T-cell clone, VASC-1, from the model. VASC-1 was determined to be a type II natural killer T-cell clone. In this study, we attempted to identify the antigen recognized by VASC-1. The monkey-derived cell line COS-7 was used because VASC-1 does not bind naturally to COS-7, although the amino acid sequences are well conserved between monkey CD1d and rat CD1d. We generated 98 COS-7 clones transfected with miscellaneous rat cDNA and screened them for VASC-1 binding. Consequently, we found one clone, 4D2, which could bind to VASC-1. Sequencing identified the rat cDNA introduced into 4D2 as sterol carrier protein 2 (SCP2). When VASC-1 was co-cultured with SCP2 knockdown rat vascular endothelial cells, VASC-1 binding was reduced significantly. Moreover, we designed a series of rat SCP2 peptides and introduced them into COS-7 cells. On the basis of VASC-1 binding and proliferation, we revealed that the peptide rSCP2(518-532) included the epitope recognized by VASC-1. Furthermore, immunization with rSCP2(518-532) accelerated the development of SVV in the rat model. The collective findings suggest that type II natural killer T cells reactive with autologous SCP2 are implicated in vascular inflammation in the rat model.

The Presence of Anti-Lactoferrin Antibodies in a Subgroup of Eosinophilic Granulomatosis with Polyangiitis Patients and Their Possible Contribution to Enhancement of Neutrophil Extracellular Trap Formation
Haruki Shida, Daigo Nakazawa, Yu Tateyama, Arina Miyoshi, Yoshihiro Kusunoki, Fumihiko Hattanda, Sakiko Masuda, Utano Tomaru, Tamihiro Kawakami, Tatsuya Atsumi, Akihiro Ishizu
FRONTIERS IN IMMUNOLOGY, 7, 636, FRONTIERS MEDIA SA, Dec. 2016, [Peer-reviewed]
English, Scientific journal, Lactoferrin (Lf) is one of the antigens of antineutrophil cytoplasmic antibodies (ANCA) and functions as an endogenous suppressor of neutrophil extracellular trap (NET) formation. However, the prevalence and pathogenicity of anti-lactoferrin antibodies (aLf) in ANCA-associated vasculitis (AAV) remain unrevealed. This study aimed to examine the significance of aLf in AAV, initially. Sixty-five sera from AAV patients, including 41 microscopic polyangiitis, 5 granulomatosis with polyangiitis, and 19 eosinophilic granulomatosis with polyangiitis (EGPA) patients, were subjected to aLf detection using enzyme-linked immunosorbent assay. Clinical characteristics were compared between aLf-positive and aLf-negative patients. Neutrophils from healthy donors were exposed to suboptimal dose (10 nM) of phorbol myristate acetate (PMA) with aLf followed by evaluation of NET formation. Results demonstrated that 4 out of 65 AAV sera (6.2%) were positive for aLf. All of them were EGPA sera (4/19, 21.1%). In EGPA, the frequency of renal involvement, serum CRP levels, and Birmingham Vasculitis Activity Score (BVAS) in the aLf-positive patients was significantly higher than those in the aLf-negative patients, and the aLf titer correlated positively with the serum CRP level and BVAS. The NET formation was particularly enhanced by combined stimulation of 10 nM PMA and 1 mu g/mL aLf. IgG isolated from sera of the aLf-positive EGPA patients (250 mu g/mL) enhanced NET formation induced by 10 nM of PMA, and the effect was abolished completely by absorption of the aLf. This pilot study suggests that aLf enhance NET formation induced by PMA and are associated with disease activity of EGPA.

膵島移植の長期生着のための課題と克服 膵島移植におけるプロテアソーム阻害による早期グラフト障害の抑制
小野 仁, 旭 よう, 吉田 雅, 腰塚 靖之, 渡辺 正明, 外丸 詩野, 江本 慎, 深井 原, 嶋村 剛, 武冨 紹信, 藤堂 省, 山下 健一郎
移植, 51, 2-3, 273, 273, (一社)日本移植学会, Aug. 2016
Japanese

NETosis markers: Quest for specific, objective, and quantitative markers
Saldko Masuda, Daigo Nakazawa, Haruki Shida, Anna Miyoshi, Yoshihiro Kusunoki, Utano Tomaru, Akihiro Ishizu
CLINICA CHIMICA ACTA, 459, 89, 93, ELSEVIER SCIENCE BV, Aug. 2016, [Peer-reviewed]
English, More than 10 years have passed since the discovery of neutrophil extracellular traps (NETs) in 2004. NETs are extracellular web-like DNA decorated with antimicrobial proteins, which are released from activated neutrophils. The state of neutrophils with NET formation is called NETosis. It has been realized that NETosis includes suicidal NETosis and vital NETosis. The former state means cell death of neutrophils, whereas the latter state preserves living neutrophilic functions. Although both suicidal and vital NETosis play essential roles in elimination of microorganisms, excessive formation of NETs, especially the ones derived from suicidal NETosis, can harm the hosts. Therefore, the discovery of NETosis markers and development of evaluation methods are important. In this review, we compare the methods for evaluating NETosis, including immunocytological and immunohistological detection of co-localized neutrophil-derived proteins and extracellular DNA, and citrullinated histones, detection of NET remnants in fluid samples, and flow cytometric detection of cell-appendant NET components, with focus on the specificity, objectivity, and quantitativity. Since the gold standard marker of NETosis or method of NET detection has not been established yet, researchers should choose the most appropriate marker or method in each situation based on the knowledge of the respective virtues and faults. (C) 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production
Yoshihiro Kusunoki, Daigo Nakazawa, Haruki Shida, Fumihiko Hattanda, Arina Miyoshi, Sakiko Masuda, Saori Nishio, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
FRONTIERS IN IMMUNOLOGY, 7, 1, 7, FRONTIERS MEDIA SA, Jun. 2016, [Peer-reviewed]
English, Scientific journal, Myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA)-associated vasculitis is a systemic small-vessel vasculitis, wherein. MPO-ANCA plays a critical role in the pathogenesis. Neutrophil extracellular traps (NETs) released from activated neutrophils are composed of extracellular web-like DNA and antimicrobial proteins, including MPO. Diverse stimuli, such as phorbol myristate acetate (PMA) and ligands of toll-like receptors (TLR), induce NETs. Although TLR-mediated NET formation can occur with preservation of living neutrophilic functions (called vital NETosis). PMA-stimulated neutrophils undergo cell death with NET formation (called suicidal NETosis). In the process of suicidal NETosis, histones are citrullinated by peptidylarginine deiminase 4 (PAD4). Since this step is necessary for decondensation of DNA. PAD4 plays a pivotal role in suicidal NETosis. Although NETs are essential for elimination of microorganisms, excessive formation of NETs has been suggested to be implicated in MPO-ANCA production. This study aimed to determine if pan-PAD inhibitors could suppress MPO-ANCA production in vivo. At first, NETs were induced in peripheral blood neutrophils derived from healthy donors (1 x 10(6)/ml) by stimulation with 20 nM PMA with or without 20 mu M propylthiouracil (PTU), an anti-thyroid drug. We then determined that the in vitro NET formation was inhibited completely by 200 mu M Cl-amidine, a pan PAD inhibitor. Next, we established mouse models with MPO-ANCA production. BALB/c mice were given intraperitoneal (i.p.) injection of PMA (50 ng at days 0 and 7) and oral PTU (2.5 mg/day) for 2 weeks. These mice were divided into two groups; the first group was given daily i.p. injection of PBS (200 mu l/day) = 13) and the other group with daily i.p. injection of Cl-amidine (0.3 mg/200 mu l PBS/day) (n = 7). Two weeks later, citrullination as an indicator of NET formation in the peritoneum and serum MPO-ANCA titer was compared between the two groups. Results demonstrated that citrullination in the peritoneum was significantly reduced in the Cl-amidine-treated mice compared with the vehicle-injected control mice (38% reduction). Additionally, the serum MPO-ANCA titer of the Cl-amidine-treated mice (32.3 +/- 31.0 ng/ml) was significantly lower than that in the vehicle-injected mice (132.1 +/- 41.6 ng/ml). The collective findings indicate that excessive formation of NETs may be implicated in MPO-ANCA production in vivo.

胆嚢悪性腫瘍が疑われたIgG4関連胆嚢炎の一例　　　　　　　　　　　　　　　
庄子 尚子, 竹元 小乃美, 渡邉 洋章, 木内 隆之, 大塚 紀幸, 和田 雅孝, 外丸 詩野, 笠原 正典
日本病理学会会誌, 105, 1, 579, 579, (一社)日本病理学会, Apr. 2016
Japanese

The responses of macrophages in interaction with neutrophils that undergo NETosis
Daigo Nakazawa, Haruki Shida, Yoshihiro Kusunoki, Arina Miyoshi, Saori Nishio, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
JOURNAL OF AUTOIMMUNITY, 67, 19, 28, ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, Feb. 2016, [Peer-reviewed]
English, Scientific journal, Neutrophil extracellular traps (NETs) are net-like chromatin fibers decorated with antimicrobial proteins, which are released from dying neutrophils. The death of neutrophils with NET formation is called NETosis. Although NETs play important roles in the innate immunity, especially in the elimination of microbes, the extracellular release of DNA and intra-cytoplasmic/nuclear proteins can, on the other hand, result in diverse adversities to the hosts. Therefore, NETosis is adequately regulated in vivo. Currently, two mechanisms, namely DNase I-dependent digestion and phagocytosis by macrophages, have been shown as such regulatory mechanisms. In this study, we focused on the interaction of macrophages and neutrophils that underwent NETosis. Results demonstrated that macrophages displayed a phenotype dependent response after degradation of NETs. Several hours after the interaction, M2 macrophages induced a pro-inflammatory response, while M1 macrophages underwent cell death with nuclear decondensation. This nuclear decondensation of M1 macrophages occurred in a peptidylarginine deiminase 4-dependent manner and resulted in a local release of extracellular DNA. Thereafter, M1 macrophages degraded DNA derived from themselves in a caspase-activated DNase-dependent manner resulting in the clearance of extracellular DNA within 24 h. This transient increase and subsequent clearance mechanism of extracellular DNA seems very reasonable in terms of the double-edged sword-like property of NETs. The collective findings demonstrate a novel phenotype- and time-dependent regulation of NETosis by macrophages. (C) 2015 The Authors. Published by Elsevier Ltd.

Novel monoclonal antibodies that recognize both rat and mouse phosphatidylserine/prothrombin complexes
Tamihiro Kawakami, Sun Young Yoon, Sora Takeuchi, Yoshinao Soma, Sayo Kuroha, Shigeru Yoshida, Haruki Shida, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
MODERN RHEUMATOLOGY, 26, 3, 470, 471, SPRINGER, 2016, [Peer-reviewed]
English

Circulating Neutrophil Extracellular Trap Levels in Well-Controlled Type 2 Diabetes and Pathway Involved in Their Formation Induced by High-Dose Glucose
Arina Miyoshi, Mai Yamada, Haruki Shida, Daigo Nakazawa, Yoshihiro Kusunoki, Akinobu Nakamura, Hideaki Miyoshi, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
PATHOBIOLOGY, 83, 5, 243, 251, KARGER, 2016, [Peer-reviewed]
English, Scientific journal, Objectives: Although intensive therapy for type 2 diabetes (T2D) prevents microvascular complications, 10% of well-controlled T2D patients develop microangiopathy. Therefore, the identification of risk markers for microvascular complications in well-controlled T2D patients is important. Recent studies have demonstrated that high-dose glucose induces neutrophil extracellular trap (NET) formation, which can be a risk for microvascular disorders. Thus, we attempted to determine the correlation of circulating NET levels with clinical/laboratory parameters in well-controlled T2D patients and to reveal the mechanism of NET formation induced by high-dose glucose. Methods: Circulating NET levels represented by myeloperoxidase (MPO)-DNA complexes in the serum of 11 well-controlled T2D patients and 13 healthy volunteers were determined by enzyme-linked immunosorbent assay. The pathway involved in the NET formation induced by high-dose glucose was determined using specific inhibitors. Results: Serum MPO-DNA complex levels were significantly higher in some well-controlled T2D patients in correlation with the clinical/laboratory parameters which have been regarded as risk markers for microvascular complications. The aldose reductase inhibitor, ranirestat, could inhibit the NET formation induced by high-dose glucose. Conclusions: Elevated levels of circulating NETs can be a risk marker for microvascular complications in well-controlled T2D patients. The polyol pathway is involved in the NET formation induced by high-dose glucose. (C) 2016 The Author(s) Published by S. Karger AG, Basel

A PROTEASOME INHIBITOR, BORTEZOMIB PREVENTS PANCREATIC ISLET GRAFT LOSS AFTER TRANSPLANTATION.
Hitoshi Ono, Yoh Asahi, Tadashi Yoshida, Yasuyuki Koshizuka, Masaaki Watanabe, Utano Tomaru, Nozomi Kobayashi, Shin Emoto, Moto Fukai, Tsuyoshi Shimamura, Akinobu Taketomi, Satoru Todo, Kenichiro Yamashita
TRANSPLANTATION, 99, 11, S122, S122, LIPPINCOTT WILLIAMS & WILKINS, Nov. 2015, [Peer-reviewed]
English

A proteasome inhibitor, bortezomib prevents pancreatic islet graft loss after transplantation
Hitoshi Ono, Yoh Asahi, Tadashi Yoshida, Yasuyuki Koshizuka, Masaaki Watanabe, Utano Tomaru, Nozomi Kobayashi, Shin Emoto, Moto Fukai, Tsuyoshi Shimamura, Akinobu Taketomi, Satoru Todo, Kenichiro Yamashita
XENOTRANSPLANTATION, 22, S75, S75, WILEY-BLACKWELL, Nov. 2015, [Peer-reviewed]
English

Fatal cardiac small-vessel involvement in ANCA-associated vasculitis: an autopsy case report
Sari Iwasaki, Akira Suzuki, Takashi Fujisawa, Taiju Sato, Shinya Shirai, Mitsunori Kamigaki, Noriyuki Otsuka, Utano Tomaru, Akihiro Ishizu
CARDIOVASCULAR PATHOLOGY, 24, 6, 408, 410, ELSEVIER SCIENCE INC, Nov. 2015, [Peer-reviewed]
English, Scientific journal, An 80-year-old Japanese man, who had fever and generalized fatigue not improved by antibiotics, was admitted to our hospital. Laboratory data indicative of renal dysfunction and antineutrophil cytoplasmic antibody (ANCA) in the serum led to the consideration of ANCA-associated vasculitis as a differential diagnosis. However, before the diagnostic confirmation, he was found dead on the bed. Autopsy revealed necrotizing crescentic glomerulonephritis in the kidneys. In addition, necrotizing granulomatous vasculitis with infiltration of multinucleated giant cells and neutrophils but not eosinophils was present in multiple organs. The direct cause of death was presumed as cardiac arrest by lethal arrhythmia because vasculitic lesions were distributed widely in the cardiac walls, acute congestion was observed in the systemic organs, and other causes of death were ruled out. This report presents the unusual manifestation of cardiac small-vessel involvement in ANCA-associated vasculitis related to sudden death. (C) 2015 Elsevier Inc. All rights reserved.

HTLV-1 Tax induces Th1 master regulator T-bet and thus IFN-gamma in CD4+CCR4+T-cells of virus-associated myelopathy patients
Araya Natsumi, Sato Tomoo, Tomaru Utano, Coler-Reilly Ariella, Yagishita Naoko, Yamauchi Junji, Hasegawa Atsuhiko, Kannagi Mari, Akiyama Hisanao, Hasegawa Yasuhiro, Takahashi Katsunori, Kunitomo Yasuo, Tanaka Yuetsu, Utsunomiya Atae, Jacobson Steven, Yamano Yoshihisa
RETROVIROLOGY, 12, 28 Aug. 2015, [Peer-reviewed]

Decreased expression of thymus-specific proteasome subunit 5t in Down syndrome patients
Utano Tomaru, Takahiro Tsuji, Shizuka Kiuchi, Akihiro Ishizu, Akira Suzuki, Noriyuki Otsuka, Tomoki Ito, Hitoshi Ikeda, Yuichiro Fukasawa, Masanori Kasahara
HISTOPATHOLOGY, 67, 2, 235, 244, WILEY-BLACKWELL, Aug. 2015, [Peer-reviewed]
English, Scientific journal, AimsThe majority of patients with Down syndrome (DS), trisomy 21, have morphologically abnormal thymuses and present with intrinsic immunological abnormalities affecting mainly the cellular immune response. The aim of this study was to examine whether the expression of functionally important molecules is altered in thymic stromal cells in patients with DS.
Methods and resultsWe analysed thymic tissues from patients with trisomy 13 (n=4), trisomy 18 (n=14) and trisomy 21 (n=13) for histological alterations, and for the expression of functionally important molecules such as 5t, a thymoproteasome subunit, and cathepsins L and S. In patients with trisomy 13 and trisomy 18, the thymus was morphologically normal or showed only mild depletion of cortical thymocytes. In contrast, the thymus showed variable histological changes in patients with trisomy 21; six of 13 cases showed severe depletion of thymocytes accompanied by the disappearance of thymic lobular architecture. In such thymuses, spindle-shaped keratin-positive cells were densely distributed, and expression of 5t, but not of cathepsin L, was markedly decreased.
ConclusionsThe present study suggests that abnormal thymic architecture and decreased expression of functionally important molecules in thymic stromal cells may be involved in immunological abnormalities in DS patients.

Presence of autoimmune regulator and absence of desmoglein 1 in a thymoma in a patient with pemphigus foliaceus.
Tsuchisaka A, Kaneko S, Imaoka K, Ota M, Kishimoto K, Tomaru U, Kasahara M, Ohata C, Furumura M, Takamori S, Morita E, Hashimoto T
The British journal of dermatology, 173, 1, 268, 271, Jul. 2015, [Peer-reviewed]

Decreased proteasomal function accelerates cigarette smoke-induced pulmonary emphysema in mice
Yosuke Yamada, Utano Tomaru, Akihiro Ishizu, Tomoki Ito, Takayuki Kiuchi, Ayako Ono, Syota Miyajima, Katsura Nagai, Tsunehito Higashi, Yoshihiro Matsuno, Hirotoshi Dosaka-Akita, Masaharu Nishimura, Soichi Miwa, Masanori Kasahara
LABORATORY INVESTIGATION, 95, 6, 625, 634, NATURE PUBLISHING GROUP, Jun. 2015, [Peer-reviewed]
English, Scientific journal, Chronic obstructive pulmonary disease (COPD) is a disease common in elderly people, characterized by progressive destruction of lung parenchyma and chronic inflammation of the airways. The pathogenesis of COPD remains unclear, but recent studies suggest that oxidative stress-induced apoptosis in alveolar cells contributes to emphysematous lung destruction. The proteasome is a multicatalytic enzyme complex that plays a critical role in proteostasis by rapidly destroying misfolded and modified proteins generated by oxidative and other stresses. Proteasome activity decreases with aging in many organs including lungs, and an age-related decline in proteasomal function has been implicated in various age-related pathologies. However, the role of the proteasome system in the pathogenesis of COPD has not been investigated. Recently, we have established a transgenic (Tg) mouse model with decreased proteasomal chymotrypsin-like activity, showing age-related phenotypes. Using this model, we demonstrate here that decreased proteasomal function accelerates cigarette smoke (CS)-induced pulmonary emphysema. CS-exposed Tg mice showed remarkable airspace enlargement and increased foci of inflammation compared with wild-type controls. Importantly, apoptotic cells were found in the alveolar walls of the affected lungs. Impaired proteasomal activity also enhanced apoptosis in cigarette smoke extract (CSE)-exposed fibroblastic cells derived from mice and humans in vitro. Notably, aggresome formation and prominent nuclear translocation of apoptosis-inducing factor were observed in CSE-exposed fibroblastic cells isolated from Tg mice. Collective evidence suggests that CS exposure and impaired proteasomal activity coordinately enhance apoptotic cell death in the alveolar walls that may be involved in the development and progression of emphysema in susceptible individuals such as the elderly.

A case of sargomatoid carcinoma of the penis
Atsushi Wanifuchi, Keisuke Taguchi, Yoshinori Ikehata, Yuichiro Kurimura, Yoshiki Hiyama, Utano Tomaru
Acta Urologica Japonica, 61, 6, 249, 252, Editorial Board of Acta Urologica Japonica, 01 Jun. 2015, [Peer-reviewed]
Japanese, Scientific journal, A 75-year-old man visited our hospital complaining of a foul smelling, painful swelling of the glans of the penis. Physical examination showed a true phimosis and a huge solid mass on the glans under the foreskin. After postectomy and penile tumor biopsy, we performed partial penectomy. Histologically, the tumor was composed of atypical spindle cells arranged in an epithelioid pattern and stained positive for both epithelial and mesenchymal markers. Therefore we diagnosed the tumor as sarcomatoid carcinoma of the penis. One month after surgery, advanced gastric cancer was discovered. Thereafter, cancer rapidly spread throughout the whole body, and he died six months postoperatively.

Establishment of a vascular endothelial cell-reactive type II NKT cell clone from a rat model of autoimmune vasculitis
Chihiro Iinuma, Masashi Waki, Ai Kawakami, Madoka Yamaguchi, Utano Tomaru, Naomi Sasaki, Sakiko Masuda, Yuki Matsui, Sari Iwasaki, Tomohisa Baba, Masanori Kasahara, Takashi Yoshiki, Daniel Paletta, Thomas Herrmann, Akihiro Ishizu
INTERNATIONAL IMMUNOLOGY, 27, 2, 105, 114, OXFORD UNIV PRESS, Feb. 2015, [Peer-reviewed]
English, Scientific journal, We previously generated a rat model that spontaneously developed small vessel vasculitis (SVV). In this study, a T cell clone reactive with rat vascular endothelial cells (REC) was established and named VASC-1. Intravenous injection of VASC-1 induced SVV in normal recipients. VASC-1 was a TCR alpha beta/CD3-positive CD4/CD8 double-negative T cell clone with expression of NKG2D. The cytokine mRNA profile under unstimulated condition was positive for IL-4 and IFN-gamma but negative for IL-2 and IL-10. After interaction with REC, the mRNA expression of IL-2, IL-5 and IL-6 was induced in VASC-1, which was inhibited by blocking of CD1d on the REC surface. Although the protein levels of these cytokines seemed to be lower than the detection limit in the culture medium, IFN-gamma was detectable. The production of IFN-gamma from the VASC-1 stimulated with LPS-pre-treated REC was inhibited by the CD1d blockade on the REC. These findings indicated VASC-1 as an NKT cell clone. The NKT cell pool includes two major subsets, namely types I and II. Type I NKT cells are characterized by expression of semi-invariant TCRs and the potential to bind to marine sponge-derived alpha-galactosylceramide (alpha-GalCer) loaded on CD1d; whereas, type II NKT cells do not manifest these characteristics. VASC-1 exhibited a usage of TCR other than the type I invariant TCR alpha chain and did not bind to alpha-GalCer-loaded CD1d; therefore, it was determined as a type II NKT cell clone. The collective evidence suggested that REC-reactive type II NKT cells could be involved in the pathogenesis of SVV in rats.

Expression of Fucosyltransferase 8 Is Associated with an Unfavorable Clinical Outcome in Non-Small Cell Lung Cancers
Rio Honma, Ichiro Kinoshita, Eiji Miyoshi, Utano Tomaru, Yoshihiro Matsuno, Yasushi Shimizu, Satoshi Takeuchi, Yuka Kobayashi, Kichizo Kaga, Naoyuki Taniguchi, Hirotoshi Dosaka-Akita
ONCOLOGY, 88, 5, 298, 308, KARGER, 2015, [Peer-reviewed]
English, Scientific journal, Objecitive: Fucosyltransferase 8 (FUT8), the only enzyme responsible for the core alpha 1,6-fucosylation of asparagine-linked oligosaccharides of glycoproteins, is a vital enzyme in cancer development and progression. We examined FUT8 expression in non-small cell lung cancers (NSCLCs) to analyze its clinical significance. We also examined the expression of guanosine diphosphate-mannose-4,6-dehydratase (GMD), which is imperative for the synthesis of fucosylated oligosaccharides. Methods: Using immunohistochemistry, we evaluated the expression of FUT8 and GMD in relation to patient survival and prognosis in potentially curatively resected NSCLCs. Results: High expression of FUT8 was found in 67 of 129 NSCLCs (51.9%) and was significantly found in non-squamous cell carcinomas (p = 0.008). High expression of FUT8 was associated with poor survival (p = 0.03) and was also a significant and independent unfavorable prognostic factor in patients with potentially curatively resected NSCLCs (p = 0.047). High expression of GMD was significantly associated with high FUT8 expression (p = 0.04). Conclusions: High expression of FUT8 is associated with an unfavorable clinical outcome in patients with potentially curatively resected NSCLCs, suggesting that FUT8 can be a prognostic factor. The analysis of FUT8 expression and its core fucosylated products may provide new insights for the therapeutic targets of NSCLCs. (C) 2015 S. Karger AG, Basel

Overexpression of TNF-alpha converting enzyme promotes adipose tissue inflammation and fibrosis induced by high fat diet
Yuki Matsui, Utano Tomaru, Arina Miyoshi, Tomoki Ito, Shinji Fukaya, Hideaki Miyoshi, Tatsuya Atsumi, Akihiro Ishizu
EXPERIMENTAL AND MOLECULAR PATHOLOGY, 97, 3, 354, 358, ACADEMIC PRESS INC ELSEVIER SCIENCE, Dec. 2014, [Peer-reviewed]
English, Scientific journal, Obesity is a state in which chronic low-grade inflammation persists in adipose tissues. Pro-inflammatory cytokines, including TNF-alpha, produced by adipose tissues have been implicated as active participants in the development of obesity-related diseases. Since TNF-alpha converting enzyme (TACE) is the major factor that induces soluble TNF-alpha, TACE has been noted as a pivotal regulator in this field. To reveal the role of TACE in adipose tissue inflammation, TACE-transgenic (TACE-Tg) and wild type (WT) mice were fed with high fat diet (HFD) or control diet for 16 weeks. At 13 weeks after the beginning of the diet, serum TNF-alpha and macrophage-related cytokine/chemokine levels were elevated in TACE-Tg mice fed with HFD (Tg-HFD mice), and the number of the so-called crown-like adipocyte was significantly increased in adipose tissues of Tg-HFD mice at the end of the experiment. Although macrophage infiltration was not detected in the adipose tissues at this time, fibrosis was observed around the crown-like adipocytes. These findings suggested that TACE overexpression induced macrophage infiltration and subsequent fibrosis in adipose tissues under HFD regimen. The collective evidence suggested that TACE could be a therapeutic target of HFD-induced obesity-related adipose tissue inflammation. (C) 2014 Elsevier Inc. All rights reserved.

Expression of α1,6-fucosyltransferase is associated with prognosis and histology in non-small cell lung cancers　　　　　　　　　　　　　　　
Honma R, Kinoshita I, Miyoshi E, Tomaru U, Matsuno Y, Shimizu Y, Takeuchi S, Kobayashi Y, Kaga K, Taniguchi N, Akita DH
Oct. 2014, [Peer-reviewed]
English, Scientific journal

HTLV-1 induces a Th1-like state in CD4+CCR4+ T cells.
Natsumi Araya, Tomoo Sato, Hitoshi Ando, Utano Tomaru, Mari Yoshida, Ariella Coler-Reilly, Naoko Yagishita, Junji Yamauchi, Atsuhiko Hasegawa, Mari Kannagi, Yasuhiro Hasegawa, Katsunori Takahashi, Yasuo Kunitomo, Yuetsu Tanaka, Toshihiro Nakajima, Kusuki Nishioka, Atae Utsunomiya, Steven Jacobson, Yoshihisa Yamano
J. Clin. Invest., 124, 8, 3431, 3442, Aug. 2014, [Peer-reviewed], [International Magazine]
English, Scientific journal, Human T-lymphotropic virus type 1 (HTLV-1) is linked to multiple diseases, including the neuroinflammatory disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T cell leukemia/lymphoma. Evidence suggests that HTLV-1, via the viral protein Tax, exploits CD4+ T cell plasticity and induces transcriptional changes in infected T cells that cause suppressive CD4+CD25+CCR4+ Tregs to lose expression of the transcription factor FOXP3 and produce IFN-γ, thus promoting inflammation. We hypothesized that transformation of HTLV-1-infected CCR4+ T cells into Th1-like cells plays a key role in the pathogenesis of HAM/TSP. Here, using patient cells and cell lines, we demonstrated that Tax, in cooperation with specificity protein 1 (Sp1), boosts expression of the Th1 master regulator T box transcription factor (T-bet) and consequently promotes production of IFN-γ. Evaluation of CSF and spinal cord lesions of HAM/TSP patients revealed the presence of abundant CD4+CCR4+ T cells that coexpressed the Th1 marker CXCR3 and produced T-bet and IFN-γ. Finally, treatment of isolated PBMCs and CNS cells from HAM/TSP patients with an antibody that targets CCR4+ T cells and i

HTLV-1 induces a Th1-like state in CD4(+)CCR4(+) T cells
Natsumi Araya, Tomoo Sato, Hitoshi Ando, Utano Tomaru, Mari Yoshida, Ariella Coler-Reilly, Naoko Yagishita, Junji Yamauchi, Atsuhiko Hasegawa, Mari Kannagi, Yasuhiro Hasegawa, Katsunori Takahashi, Yasuo Kunitomo, Yuetsu Tanaka, Toshihiro Nakajima, Kusuki Nishioka, Atae Utsunomiya, Steven Jacobson, Yoshihisa Yamano
JOURNAL OF CLINICAL INVESTIGATION, 124, 8, 3431, 3442, AMER SOC CLINICAL INVESTIGATION INC, Aug. 2014, [Peer-reviewed]
English, Scientific journal, Human T-Iymphotropic virus type 1 (HTLV-1) is linked to multiple diseases, including the neuroinflammatory disease HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T cell leukemia/lymphoma. Evidence suggests that HTLV-1, via the viral protein Tax, exploits CD4(+)T cell plasticity and induces transcriptional changes in infected T cells that cause suppressive CD4(+)CD25(+)CCR4(+) Tregs to lose expression of the transcription factor FOXP3 and produce IFN-gamma, thus promoting inflammation. We hypothesized that transformation of HTLV-1 infected CCR4(+)T cells into Th1-like cells plays a key role in the pathogenesis of HAM/TSP. Here, using patient cells and cell lines, we demonstrated that Tax, in cooperation with specificity protein 1 (Sp1), boosts expression of the Th1 master regulator T box transcription factor (T-bet) and consequently promotes production of IFN-gamma. Evaluation of CSF and spinal cord lesions of HAM/TSP patients revealed the presence of abundant CD4(+)CCR4(+)T cells that coexpressed the Th1 marker CXCR3 and produced T-bet and IFN-gamma. Finally, treatment of isolated PBMCs and CNS cells from HAM/TSP patients with an antibody that targets CCR+T cells and induces cytotoxicity in these cells reduced both viral load and IFN-gamma production, which suggests that targeting CCR4(+)T cells may be a viable treatment option for HAM/TSP.

Decreased proteasomal activity causes photoreceptor degeneration in mice
Ryo Ando, Kousuke Noda, Utano Tomaru, Mamoru Kamoshita, Yoko Ozawa, Shoji Notomi, Toshio Hisatomi, Mika Noda, Atsuhiro Kanda, Tatsuro Ishibashi, Masanori Kasahara, Susumu Ishida
Investigative Ophthalmology and Visual Science, 55, 7, 4682, 4690, Association for Research in Vision and Ophthalmology Inc., 03 Jul. 2014, [Peer-reviewed]
English, Scientific journal, Purpose: To study the retinal degeneration caused by decreased proteasomal activity in β5t transgenic (β5t-Tg) mice, an animal model of senescence acceleration. Methods: β5t-Tg mice and age-matched littermate control (WT) mice were used. Proteasomal activities and protein level of poly-ubiquitinated protein in retinal extracts were quantified. Fundus images of β5t-Tg mice were taken and their features were assessed. For histologic evaluation, the thicknesses of inner nuclear layer (INL), outer nuclear layer (ONL), and photoreceptor outer segment (OS) were measured. For functional analysis, ERG was recorded under scotopic and photopic illumination conditions. Immunofluorescence (IF) staining and TUNEL were performed to investigate the mechanism of photoreceptor degeneration. Results: Chymotrypsin-like activity was partially suppressed in retinal tissues of β5t-Tg mice. Retinal degenerative changes with arterial attenuation were present in β5t-Tg, but not in WT mice. Inner nuclear layer thickness showed no significant change between β5t-Tg and WT mice at 1, 3, 6, and 9 months of age. By contrast, thicknesses of ONL and OS in β5t-Tg mice were significantly decreased at 3, 6, and 9 months compared with those in WT mice. Electroretinograms showed decrease of scotopic a-wave amplitude in β5t-Tg mice. The number of TUNEL-positive cells in ONL were significantly increased in β5t-Tg mice and colocalized with apoptosis-inducing factor, but not with cleaved caspase-3 and -9, indicating that the photoreceptor cell death was induced via a caspase-independent pathway. Conclusions: The current data showed that impaired proteasomal function causes photoreceptor degeneration. © 2014 The Association for Research in Vision and Ophthalmology, Inc.

Enhanced Formation and Disordered Regulation of NETs in Myeloperoxidase-ANCA-Associated Microscopic Polyangiitis
Daigo Nakazawa, Haruki Shida, Utano Tomaru, Masaharu Yoshida, Saori Nishio, Tatsuya Atsumi, Akihiro Ishizu
JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 25, 5, 990, 997, AMER SOC NEPHROLOGY, May 2014, [Peer-reviewed]
English, Scientific journal, AKI involves early Toll-like receptor (TLR)-driven immunopathology, and resolution of inflammation is needed for rapid regeneration of injured tubule cells. Notably, activation of TLRs also has been implicated in epithelial repair. We hypothesized that TLR signaling drives tubule regeneration after acute injury through the induction of certain ILs. Systematic screening in vitro identified IL-22 as a candidate proregeneratory factor in primary tubular cell recovery, and IL-22 deficiency or IL-22 blockade impaired post-ischemic tubular recovery after AKI in mice. Interstitial mononuclear cells, such as dendritic cells and macrophages, were the predominant source of IL-22 secretion, whereas IL-22 receptor was expressed by tubular epithelial cells exclusively. Depleting IL-22-producing cells during the healing phase impaired epithelial recovery, which could be rescued entirely by reconstituting mice with IL-22. In vitro, necrotic tubular cells and oxidative stress induced IL-22 secretion selectively through TLR4. Although TLR4 blockade during the early injury phase prevented tubular necrosis and AKI, TLR4 blockade during the healing phase suppressed IL-22 production and impaired kidney regeneration. Taken together, these results suggest that necrotic cell-derived TLR4 agonists activate intrarenal mononuclear cells to secrete IL-22, which accelerates tubular regeneration and recovery in AKI.

Corticomedullary differentiation and maturational arrest in thymomas
Philipp Stroebel, Elena Hartmann, Andreas Rosenwald, Joerg Kalla, German Ott, Godehard Friedel, Berthold Schalke, Masanori Kasahara, Utano Tomaru, Alexander Marx
HISTOPATHOLOGY, 64, 4, 557, 566, WILEY-BLACKWELL, Mar. 2014, [Peer-reviewed]
English, Scientific journal, AimsMorphological complexity hampers the histological classification of thymomas. Our aim was to determine whether the use of novel differentiation and maturation markers of cortical and medullary thymic epithelial cells (cTECs and mTECs) might provide an approach to understanding the underlying biology of these tumours.
Methods and resultsFifty-seven thymomas were studied by immunohistochemistry. The cortical markers used were B5T, PRSS16, and cathepsin V. The medullary markers used were CD40, claudin-4, AIRE, and desmin. Involucrin and cytokeratin 10 were used to study terminal mTEC maturation. Irrespective of histological subtype, most thymomas contained distinct areas with cortical and medullary differentiation. Type B1, type B2 and type AB thymomas showed marked bi-lineage differentiation, with lack of terminal mTEC maturation in type AB. Type AB thymomas were unique in showing areas where cells with either cortical or medullary differentiation were intimately mixed' at the single-cell level. Type B3 and type A thymomas showed only abortive lineage differentiation and maturation.
ConclusionsThymomas show highly characteristic patterns of bi-lineage TEC differentiation that reflect the histological subtypes recognized by the WHO classification. We hypothesize that thymomas arise from thymic precursor cells with different cortical and/or medullary maturation defects.

Expression of thymoproteasome subunit beta 5t in type AB thymoma
Yosuke Yamada, Utano Tomaru, Akihiro Ishizu, Takayuki Kiuchi, Masanori Kasahara, Yoshihiro Matsuno
JOURNAL OF CLINICAL PATHOLOGY, 67, 3, 276, 278, BMJ PUBLISHING GROUP, Mar. 2014, [Peer-reviewed]
English, Scientific journal, Type AB thymoma is a thymic epithelial tumour composed of lymphocyte-poor type A and lymphocyterich type B components. Although it is categorised as a single entity in the classification of WHO, it shows a broad range of morphology. To investigate whether the functional characteristic of neoplastic cells in type AB thymoma relates to morphological diversity, we performed immunohistochemical analysis using anti-beta 5t antibody in 20 cases of type AB thymoma. beta 5t is a recently discovered proteasomal beta subunit expressed exclusively in cortical thymic epithelial cells and tumour epithelial cells of thymomas with cortical differentiation. Consistent with our previous observation, beta 5t was predominantly expressed in the type B component. When the type B component was divided into three groups morphologically, beta 5t was expressed more frequently in cases with round to polygonal than spindle to oval tumour cells. Furthermore, the ratio of terminal deoxynucleotidyl transferase (TdT)-positive lymphocytes was increased in components with higher expression of beta 5t. These results indicate that the histological diversity of type AB thymoma correlates with expression of a functional marker beta 5t and abundance of TdT-positive lymphocytes.

Serum level of soluble triggering receptor expressed on myeloid cells-1 as a biomarker of disease activity in relapsing polychondritis
Tomoo Sato, Yoshihisa Yamano, Utano Tomaru, Yukiko Shimizu, Hitoshi Ando, Takahiro Okazaki, Hiroko Nagafuchi, Jun Shimizu, Shoichi Ozaki, Teruomi Miyazawa, Kazuo Yudoh, Hiroshi Oka, Noboru Suzuki
MODERN RHEUMATOLOGY, 24, 1, 129, 136, SPRINGER, Jan. 2014, [Peer-reviewed]
English, Scientific journal, Objectives We aimed to identify a serum biomarker for evaluating the disease activity of relapsing polychondritis (RP).
Methods We measured and compared serum levels of 28 biomarkers potentially associated with this disease, including soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), high-sensitivity C-reactive protein (hs-CRP), and cartilage oligomeric matrix protein (COMP), in 15 RP patients and 16 healthy donors (HDs). We divided the 15 RP patients into active RP (n = 8) and inactive RP (n = 7) groups, depending on the extent of the disease, and compared candidate markers between groups. The localization of membrane-bound TREM-1 in the affected tissue was examined by immunohistochemistry.
Results Serum levels of sTREM-1, interferon-gamma, chemokine (C-C motif) ligand 4, vascular endothelial growth factor, and matrix metalloproteinases-3 were significantly higher in RP patients than HDs. Among these markers, sTREM-1 had the highest sensitivity and specificity (86.7 and 86.7 %, respectively). Furthermore, the serum level of sTREM-1 was significantly higher in active RP patients than inactive RP patients (p = 0.0403), but this was not true for hs-CRP or COMP. TREM-1 was expressed on endothelial cells in RP lesions.
Conclusions The serum level of sTREM-1 may be a useful marker of disease activity in RP.

Possible linkage between microscopic polyangiitis and thrombosis via neutrophil extracellular traps
T. Imamoto, D. Nakazawa, H. Shida, A. Suzuki, N. Otsuka, U. Tomaru, A. Ishizu
CLINICAL AND EXPERIMENTAL RHEUMATOLOGY, 32, 1, 149, 150, CLINICAL & EXPER RHEUMATOLOGY, Jan. 2014, [Peer-reviewed]
English

Ag and IL-2 immune complexes efficiently expand Ag-specific Treg cells that migrate in response to chemokines and reduce localized immune responses
Ryoko Hamano, Tomohisa Baba, Soichiro Sasaki, Utano Tomaru, Akihiro Ishizu, Mitsuhiro Kawano, Masakazu Yamagishi, Naofumi Mukaida
European Journal of Immunology, 44, 4, 1005, 1015, Wiley-VCH Verlag, 2014, [Peer-reviewed]
English, Scientific journal, An intravenous administration of a high-dose antigen (Ag) can induce immune tolerance and suppress the immune response, but the mechanism remains unclear. We recently proved that a combined i.v. administration of OVA and IL-2-anti-IL-2 Ab immune complexes (IL-2 ICs) efficiently expands OVA-specific Treg cells in the thymus and induces their migration into peripheral blood, by using OVA-specific TCR Tg-expressing DO11.10 mice. Here, we demonstrate that the expanded OVA-specific Treg cells rapidly move into the air pouch after OVA injection in DO11.10 mice. The migration was inhibited by blocking the axis of a chemokine receptor, CCR2. Moreover, prior treatment with OVA and IL-2 ICs enhanced OVA-specific Treg-cell migration and inhibited OVA-induced delayed-type hypersensitivity (DTH) reactions in the skin of BM chimeric mice with 15% of T cells expressing OVA-specific TCR. Blocking the CCR2 axis reversed this suppression of DTH in these mice. Furthermore, prior treatment with OVA and IL-2 ICs effectively reduced DTH reactions even in WT mice possessing only a very small population of OVA-specific T cells. Thus, the treatment with Ag and IL-2 ICs can efficiently expand Ag-specific Treg cells with the capacity to migrate and reduce localized immune responses. © 2013 WILEY-VCH Verlag GmbH &
Co. KGaA, Weinheim.

[NETs in pathogenesis of vasculitis].
Nakazawa D, Nishio S, Tomaru U, Atsumi T, Ishizu A
Nihon Jinzo Gakkai shi, 56, 2, 117, 123, 2014, [Peer-reviewed]

Pulmonary veno-occlusive disease(PVOD)の一剖検例　　　　　　　　　　　　　　　
山田 洋介, 大塚 紀幸, 大平 洋, 辻野 一三, 深谷 進司, 外丸 詩野, 石津 明洋
脈管学, 53, December, 242, 243, (一社)日本脈管学会, Dec. 2013, [Peer-reviewed]
Japanese

Possible implication of disordered neutrophil extracellular traps in the pathogenesis of MPO-ANCA-associated vasculitis
Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
CLINICAL AND EXPERIMENTAL NEPHROLOGY, 17, 5, 631, 633, SPRINGER, Oct. 2013, [Peer-reviewed]
English, Neutrophil extracellular traps (NETs) are characterized by the presence of extracellular DNA fibers studded with antimicrobial proteins, including myeloperoxidase (MPO). Although NETs play an important role in the innate immune system, the scattered extracellular enzymes, such as MPO, pose risks to the host. Therefore, NETs are strictly regulated by DNase I in the serum, which prevents them from persisting. Recent studies have demonstrated that dysregulation of NETs could be involved in the pathogenesis of autoimmune diseases, including systemic lupus erythematosus. In this review, we interpret the association of disordered NETs with autoimmune diseases, especially propylthiouracil-induced MPO-ANCA-associated vasculitis.

Thymoproteasome: Role in thymic selection and clinical significance as a diagnostic marker for thymic epithelial tumors
Utano Tomaru, Masanori Kasahara
Archivum Immunologiae et Therapiae Experimentalis, 61, 5, 357, 365, Oct. 2013, [Peer-reviewed]
English, The thymoproteasome is a specialized type of proteasomes expressed exclusively in the thymic cortex. It has a unique catalytic subunit β5t with unusual enzymatic activity. The thymoproteasome exhibits lower chymotrypsin-like activity than other forms of proteasomes such as constitutive proteasomes and immunoproteasomes. Its cleavage specificity appears uniquely suited for the production of peptides that mediate positive selection of CD8+ T cells. Similar to major histocompatibility complex molecules and T/B-cell receptors, the thymoproteasome occurs only in jawed vertebrates, suggesting that it evolved concomitant with the cardinal elements of adaptive immunity. β5t can be used as a marker in the differential diagnosis of thymic tumors. It is expressed in most type B and some type AB thymomas, but not in type A thymoma, thymic carcinoma, or tumors of non-thymic epithelial origin. © 2013 L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.

Lysosomal-associated membrane protein-2 plays an important role in the pathogenesis of primary cutaneous vasculitis
Sora Takeuchi, Satoko Kimura, Yoshinao Soma, Masashi Waki, Madoka Yamaguchi, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu, Tamihiro Kawakami
RHEUMATOLOGY, 52, 9, 1592, 1598, OXFORD UNIV PRESS, Sep. 2013, [Peer-reviewed]
English, Scientific journal, Methods. Cutaneous vasculitis was observed in similar to 30% of 6-month-old morbid env-pX rats. In contrast, these findings were rare in premorbid env-pX rats under 3 months old. We also examined 85 patients with CPN and 36 adult patients with HSP. Serum anti-LAMP-2 antibody levels were determined using ELISA. Premorbid env-pX rats under 3 months old were given an i.v. injection of anti-LAMP-2 antibody at day 0 and day 7. At day 14, these rats underwent histopathological and direct immunofluorescence examination. Cell surface LAMP-2 expression of rat neutrophils was examined by flow cytometry.
Results. Serum anti-LAMP-2 antibody levels were significantly higher in morbid env-pX rats than in wild-type normal rats. In addition, the levels in the cutaneous vasculitis group of morbid env-pX rats were significantly higher than the no cutaneous vasculitis group. Intravenous anti-LAMP-2 antibody injection into premorbid env-pX rats under 3 months old induced infiltration of neutrophils into cutaneous small vessels. Anti-LAMP-2 antibody-binding neutrophils were detected there. LAMP-2 expression on the cell surface of neutrophils in premorbid env-pX rats under PMA stimulation was higher compared with controls. Serum anti-LAMP-2 antibody levels in CPN and HSP were significantly higher than those of healthy controls.
Conclusion. These data support a positive relationship between anti-LAMP-2 antibody and cutaneous vasculitis.

Positive feedback loop via astrocytes causes chronic inflammation in virus-associated myelopathy
Hitoshi Ando, Tomoo Sato, Utano Tomaru, Mari Yoshida, Atae Utsunomiya, Junji Yamauchi, Natsumi Araya, Naoko Yagishita, Ariella Coler-Reilly, Yukiko Shimizu, Kazuo Yudoh, Yasuhiro Hasegawa, Kusuki Nishioka, Toshihiro Nakajima, Steven Jacobson, Yoshihisa Yamano
BRAIN, 136, Pt 9, 2876, 2887, OXFORD UNIV PRESS, Sep. 2013, [Peer-reviewed]
English, Scientific journal, Human T-lymphotropic virus type 1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a rare neurodegenerative disease characterized by chronic inflammation in the spinal cord. We hypothesized that a positive feedback loop driven by chemokines may be responsible for the chronic inflammation in HAM/TSP. We aimed to determine the identity of these chemokines, where they are produced, and how they drive chronic inflammation in HAM/TSP. We found that patients with HAM/TSP have extraordinarily high levels of the chemokine CXCL10 (also known as IP-10) and an abundance of cells expressing the CXCL10-binding receptor CXCR3 in the cerebrospinal fluid. Histological analysis revealed that astrocytes are the main producers of CXCL10 in the spinal cords of patients with HAM/TSP. Co-culture of human astrocytoma cells with CD4(+) T cells from patients with HAM/TSP revealed that astrocytes produce CXCL10 in response to IFN-gamma secreted by CD4(+) T cells. Chemotaxis assays results suggest that CXCL10 induces migration of peripheral blood mononuclear cells to the central nervous system and that anti-CXCL10 neutralizing antibody can disrupt this migration. In short, we inferred that human T-lymphotropic virus type 1-infected cells in the central nervous system produce IFN-gamma that induces astrocytes to secrete CXCL10, which recruits more infected cells to the area via CXCR3, constituting a T helper type 1-centric positive feedback loop that results in chronic inflammation.

Human papillomavirus 16-positive uterine cervical squamous cell carcinoma with coinfection with human papillomavirus 34 has a lower incidence in lymph node metastasis than that without coinfection with human papillomavirus 34
Rie Michimata, Hidemichi Watari, Utano Tomaru, Noriaki Sakuragi, Akihiro Ishizu
Pathobiology, 80, 5, 259, 264, Jun. 2013, [Peer-reviewed]
English, Scientific journal, Our earlier study demonstrated high prevalence of multiple human papillomavirus (HPV) infection in patients with invasive uterine cervical cancer, including squamous cell carcinoma (SCC). HPV 16 is the most predominant genotype related to SCC of the uterine cervix. The aim of this study was to reveal the biological significance of multiple HPV infection concerning the tumor progression of invasive uterine cervical SCC. In the present study, the effects of coinfection with genotypes other than HPV 16 on tumor growth and lymph node metastasis of invasive uterine cervical SCC with HPV 16 infection were examined. Although coinfection with most genotypes did not influence tumor progression, the clinical stage of patients coinfected with HPV 16 and HPV 34 was significantly lower than that of those without HPV 34 coinfection (p = 0.0038). Moreover, no patient coinfected with HPV 16 and HPV 34 manifested lymph node metastasis, but about half of the patient population without HPV 34 coinfection did (p = 0.0299). These findings suggested that coinfection with HPV 34 could prevent the tumor progression of invasive uterine cervical SCC with HPV 16 infection. Copyright © 2013 S. Karger AG, Basel.

[Successful rituximab treatment for acquired amegakaryocytic thrombocytopenic purpura complicated with Coombs-negative autoimmune hemolytic anemia].
Hashimoto A, Fujimi A, Kanisawa Y, Matsuno T, Okuda T, Minami S, Doi T, Ishikawa K, Uemura N, Tomaru U
[Rinsho ketsueki] The Japanese journal of clinical hematology, 54, 6, 568, 573, The Japanese Society of Hematology, Jun. 2013, [Peer-reviewed]
Japanese, Acquired amegakaryocytic thrombocytopenic purpura (AATP) is a rare disorder characterized by severe thrombocytopenia associated with total absence or a selective decrease in bone marrow megakaryocytes. A 67-year-old male presented with a 2-month bleeding tendency. He was referred to our hospital because of severe thrombocytopenia. Bone marrow biopsy showed complete absence of megakaryocytes without dysplasia in cells of the myeloid and erythroid lineages. AATP was diagnosed. In addition, mild normocytic normochromic anemia and reticulocytosis were also observed and haptoglobin was below the detectable level. Coombs-negative autoimmune hemolytic anemia (AIHA) was diagnosed based on the high titer of RBC-bound IgG and negative direct and indirect coombs test results. He was first treated with cyclosporine 200 mg per day and subsequently with prednisolone but only slight temporary improvement was achieved. Administration of eight doses of rituximab 375 mg/m² per week ameliorated both thrombocytopenia and anemia. AATP should be considered in the differential diagnosis of thrombocytopenia, and immunosuppressive therapy is a potential first-line treatment. This is the first case report of AATP accompanied by AIHA successfully treated with rituximab.

Protective Roles of Epithelial Cells in the Survival of Adult T-Cell Leukemia/Lymphoma Cells
Yukiko Miyatake, Andre L. A. Oliveira, Mohamed Ali Jarboui, Shuichi Ota, Utano Tomaru, Takanori Teshima, William W. Hall, Masanori Kasahara
AMERICAN JOURNAL OF PATHOLOGY, 182, 5, 1832, 1842, ELSEVIER SCIENCE INC, May 2013, [Peer-reviewed]
English, Scientific journal, Adult T-cell leukemia/lymphoma (ATL) is a highly invasive and intractable T-cell malignancy caused by human T-cell leukemia virus-1 infection. We demonstrate herein that normal tissue-derived epithelial cells (NECs) exert protective effects on the survival of leukemic cells, which may partially account for high resistance to antileukemic therapies in patients with ATL. Viral gene-silenced, ATL-derived cell Lines (ATL cells) dramatically escaped from histone deacetylase inhibitor-induced apoptosis by direct co-culture with NECs. Adhesions to NECs suppressed p21(Cip1) expression and increased a proportion of resting G0/G1 phase cells in trichostatin A (TSA)-treated ATL cells. ATL cells adhering to NECs down-regulated CD25 expression and enhanced vimentin expression, suggesting that most ATL cells acquired a quiescent state by cell-cell interactions with NECs. ATL cells adhering to NECs displayed highly elevated expression of the cancer stem cell marker CD44. Blockade of CD44 signaling diminished the NEC-conferred resistance of ATL cells to TSA-induced apoptosis. Co-culture with NECs also suppressed the expression of NKG2D Ligands on TSA-treated ATL cells, resulting in decreased natural killer cell-mediated cytotoxicity. Combined evidence suggests that interactions with normal epithelial cells augment the resistance of ATL cells to TSA-induced apoptosis and facilitate immune evasion by ATL cells.

Prediction of Response to Treatment by Gene Expression Profiling of Peripheral Blood in Patients with Microscopic Polyangiitis
Akihiro Ishizu, Utano Tomaru, Taichi Murai, Tomohiro Yamamoto, Tatsuya Atsumi, Takashi Yoshiki, Wako Yumura, Kunihiro Yamagata, Hidehiro Yamada, Shunichi Kumagai, Manae S. Kurokawa, Machi Suka, Hirofumi Makino, Shoichi Ozaki
PLOS ONE, 8, 5, e63182, PUBLIC LIBRARY SCIENCE, May 2013, [Peer-reviewed]
English, Scientific journal, The JMAAV study was an open-labeled prospective clinical trial, which proposed severity-based treatment protocols for patients with microscopic polyangiitis (MPA). The results suggest that the proposed protocols are useful (remission rate: 89.4%), but are also indicative of relapse or patient demise regardless of the treatment (recurrence rate: 19.0%; mortality rate: 10.6%). The aim of this study is to develop the method to predict response to the treatment in patients with MPA. In the present study, transcriptome analysis was performed using peripheral blood from patients enrolled in the JMAAV study before and 1-week after the beginning of treatment. The gene expression profile before treatment was not directly related to the response to the treatment. However, when the samples from 9 patients with good response (persistent remission for 18 months) were examined, the expression of 88 genes was significantly altered by the treatment. Thirty statistically reliable genes were selected, and then the alteration of expression by the treatment was examined among 22 patients, including 17 with good response, which was defined as persistent remission for 18 months and 5 with poor response, which was defined as relapse after remission or no remission. Discrimination analysis between the alteration of expression of the 30 genes by the treatment and the response identified a combination of 16 genes as the most valuable gene set to predict the response to the treatment. This preliminary study identified IRF7, IFIT1, IFIT5, OASL, CLC, GBP-1, PSMB9, HERC5, CCR1, CD36, MS4A4A, BIRC4BP, PLSCR1, DEFA1/DEFA3, DEFA4, and COL9A2 as the important genes that can predict the response to the treatment in patients with MPA at an early point during the therapy.

Serum level of soluble triggering receptor expressed on myeloid cells-1 as a biomarker of disease activity in relapsing polychondritis.
Sato T, Yamano Y, Tomaru U, Shimizu Y, Ando H, Okazaki T, Nagafuchi H, Shimizu J, Ozaki S, Miyazawa T, Yudoh K, Oka H, Suzuki N
Modern rheumatology / the Japan Rheumatism Association, Feb. 2013, [Peer-reviewed]

Overexpression of TNF-alpha-converting enzyme in fibroblasts augments dermal fibrosis after inflammation
Shinji Fukaya, Yuki Matsui, Utano Tomaru, Ai Kawakami, Sayuri Sogo, Toshiyuki Bohgaki, Tatsuya Atsumi, Takao Koike, Masanori Kasahara, Akihiro Ishizu
LABORATORY INVESTIGATION, 93, 1, 72, 80, NATURE PUBLISHING GROUP, Jan. 2013, [Peer-reviewed]
English, Scientific journal, TNF-alpha-converting enzyme (TACE) can cleave transmembrane proteins, such as TNF-alpha, TNF receptors, and epidermal growth factor receptor (EGFR) ligands, to release the extracellular domains from the cell surface. Recent studies have suggested that overexpression of TACE may be associated with the pathogenesis of inflammation and fibrosis. To determine the roles of TACE in inflammation and fibrosis, TACE transgenic (TACE-Tg) mice, which overexpressed TACE systemically, were generated. As the transgene-derived TACE was expressed as an inactive form, no spontaneous phenotype developed in TACE-Tg mice. However, the transgene-derived TACE could be converted to an active form by furin in vitro and by phorbol myristate acetate (PMA) in vivo. Subcutaneous injection of PMA into mice induced inflammatory cell infiltration 1 day later and subsequent dermal fibrosis 7 days later. Interestingly, the degree of dermal fibrosis at day 7 was significantly higher in TACE-Tg mice than in wild-type mice. Correspondingly, PMA increased the expression of type I collagen in the primary culture of dermal fibroblasts derived from TACE-Tg mice. Furthermore, phosphorylated EGFR was increased in the fibroblasts by the PMA treatment. The collective findings suggest that TACE overexpression and activation in fibroblasts could shed off putative EGFR ligands. Subsequently, the soluble EGFR ligands could bind and activate EGFR on fibroblasts, and then increase the type I collagen expression resulting in induction of dermal fibrosis. These results also suggest that TACE and EGFR on fibroblasts may be novel therapeutic targets of dermal fibrosis, which is induced after diverse inflammatory disorders of the skin. Laboratory Investigation (2013) 93, 72-80; doi:10.1038/labinvest.2012.153; published online 12 November 2012

Possible Implication of Fc gamma Receptor-Mediated Trogocytosis in Susceptibility to Systemic Autoimmune Disease
Sakiko Masuda, Sari Iwasaki, Utano Tomaru, Tomohisa Baba, Kazuaki Katsumata, Akihiro Ishizu
CLINICAL & DEVELOPMENTAL IMMUNOLOGY, 2013, 345745, HINDAWI PUBLISHING CORPORATION, 2013, [Peer-reviewed]
English, Leukocytes can "gnaw away" the plasma membrane of other cells. This phenomenon, called trogocytosis, occurs subsequent to cell-to-cell adhesion. Currently, two mechanisms of trogocytosis, adhesion molecule-mediated trogocytosis and Fc gamma receptor-(Fc gamma R-) mediated trogocytosis, have been identified. In our earlier study, we established an in vitro model of Fc gamma R-mediated trogocytosis, namely, CD8 translocationmodel from T cells to neutrophils. By using this model, we demonstrated that the molecules transferred to neutrophils via Fc gamma R-mediated trogocytosis were taken into the cytoplasm immediately. This result suggests that the chance of molecules transferred via Fc gamma R-mediated trogocytosis to play a role on the cell surface could be time-limited. Thus, we consider the physiological role of Fc gamma R-mediated trogocytosis as a means to remove antibodies (Abs) that bind with self-molecules rather than to extractmolecules from other cells. This concept means that Fc gamma R-mediated trogocytosis can be a defense mechanism to Ab-mediated autoimmune response. Moreover, the activity of Fc gamma R-mediated trogocytosis was revealed to be parallel to the endocytotic activity of neutrophils, which was critically related to the susceptibility to systemic autoimmune diseases. The collective findings suggest that Fc gamma R-mediated trogocytosis could physiologically play a role in removal of Abs bound to self-antigens and prevent autoimmune diseases.

Primary diffuse large B-cell lymphoma of the uterine cervix successfully treated with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone chemotherapy - A case report
Akari Hashimoto, Akihito Fujimi, Yuji Kanisawa, Teppei Matsuno, Toshinori Okuda, Shinya Minami, Tadashi Doi, Kazuma Ishikawa, Naoki Uemura, Yuko Jyomen, Utano Tomaru
Japanese Journal of Cancer and Chemotherapy, 40, 13, 2589, 2592, Japanese Journal of Cancer and Chemotherapy Publishers Inc., 2013, [Peer-reviewed]
Japanese, Scientific journal, Primary malignant lymphoma of the uterine cervix is a rare disease, and the therapeutic strategy has not been clearly established. A 45-year old woman presented with vaginal bleeding and hypermenorrhea in January 2012. Physical examination revealed a mass in the pelvic cavity approximately the size of a neonate's head. Pelvic magnetic resonance imaging (MRI) showed a solid mass 11 cm in size in the uterine cervix with homogeneous low intensity on T1-weighted images, iso-high intensity on T2-weighted images, and heterogeneous iso-high intensity on gadolinium-diethylenetriaminepentaacetate (Gd-DTPA)-enhanced images. Multiple lymphadenopathy were also detected in the pelvis. The Papanicolaou smear indicated class 5 cervical cytology, and a subsequent histological examination by a punch biopsy of the cervix showed diffuse infiltration of medium- to large-sized mononuclear cells that stained positive for CD20 and CD79a and negative for CD3, CD5, and EBER. Bone marrow biopsy revealed no abnormality. Positron emission tomography-computed tomography (PET-CT) showed strong fluorodeoxyglucose (FDG) accumulation in the uterine cervix mass, and in the pelvic and right inguinal lymphadenopathy. The patient was diagnosed with diffuse large B-cell lymphoma of the uterine cervix, Ann Arbor stage II AE. She was successfully treated with 8 courses of rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) chemotherapy, and maintains a complete remission.

肉芽腫性乳腺炎の1例
飯村 泰昭, 岩井 和浩, 狭間 一明, 浅野 賢道, 佐藤 暢人, 京極 典憲, 大塚 紀幸, 外丸 詩野
北海道外科雑誌, 57, 2, 195, 195, 北海道外科学会, Dec. 2012
Japanese

Mechanism of Fc gamma Receptor-Mediated Trogocytosis-Based False-Positive Results in Flow Cytometry
Sakiko Masuda, Sari Iwasaki, Utano Tomaru, Juri Sato, Ai Kawakami, Kana Ichijo, Sayuri Sogo, Tomohisa Baba, Kazuaki Katsumata, Masanori Kasahara, Akihiro Ishizu
PLOS ONE, 7, 12, e52918, PUBLIC LIBRARY SCIENCE, Dec. 2012, [Peer-reviewed]
English, Scientific journal, The whole blood erythrocyte lysis method is the most common protocol of sample preparation for flow cytometry (FCM). Although this method has many virtues, our recent study has demonstrated false-positive results when surface markers of monocytes were examined by this method due to the phenomenon called Fc gamma receptor (Fc gamma R)-mediated trogocytosis. In the present study, similar Fc gamma R-mediated trogocytosis-based false-positive results have been demonstrated when granulocytes were focused on instead of monocytes. These findings indicated that not only monocytes but also granulocytes, the largest population with Fc gamma R expression in peripheral blood, could perform Fc gamma R-mediated trogocytosis. Since the capacity of Fc gamma R-mediated trogocytosis was different among blood samples, identification of factors that could regulate the occurrence of Fc gamma R-mediated trogocytosis should be important for the quality control of FCM. Our studies have suggested that such factors are present in the serum. In order to identify the serum factors, we employed the in vitro model of Fc gamma R-mediated trogocytosis using granulocytes. Investigation with this model determined the serum factors as heat-labile molecules with molecular weight of more than 100 kDa. Complements in the classical pathway were initially assumed as candidates; however, the C1 inhibitor did not yield an obvious influence on Fc gamma R-mediated trogocytosis. On the other hand, although immunoglobulin ought to be resistant to heat inactivation, the inhibitor of human anti-mouse antibodies (HAMA) effectively blocked Fc gamma R-mediated trogocytosis. Moreover, the inhibition rates were significantly higher in HAMA(high) serum than HAMA(low) serum. The collective findings suggested the involvement of heterophilic antibodies such as HAMA in the mechanism of false-positive results in FCM due to Fc gamma R-mediated trogocytosis.

Abnormal conformation and impaired degradation of propylthiouracil-induced neutrophil extracellular traps Implications of disordered neutrophil extracellular traps in a rat model of myeloperoxidase antineutrophil cytoplasmic antibody-associated vasculitis
Daigo Nakazawa, Utano Tomaru, Akira Suzuki, Sakiko Masuda, Risa Hasegawa, Toshiaki Kobayashi, Saori Nishio, Masanori Kasahara, Akihiro Ishizu
ARTHRITIS AND RHEUMATISM, 64, 11, 3779, 3787, WILEY-BLACKWELL, Nov. 2012, [Peer-reviewed]
English, Scientific journal, Objective Neutrophil extracellular traps (NETs) are composed of DNA and antimicrobial proteins, including myeloperoxidase (MPO). Recent studies have demonstrated that impaired regulation of NETs could trigger an autoimmune response. Propylthiouracil (PTU), an antithyroid drug, is associated with a risk of MPO antineutrophil cytoplasmic antibody (ANCA) production and MPO ANCA-associated vasculitis (MPO AAV). This study was undertaken to clarify the mechanism of MPO ANCA production, using the PTU-induced model of MPO AAV.
Methods NETs were induced by treating human neutrophils with phorbol myristate acetate (PMA) in vitro. We examined whether the addition of PTU influenced the NET formation induced by PMA and the degradation of NETs by DNase I, which is regarded as a regulator of NETs. Furthermore, we examined whether NETs generated by the combination of PMA and PTU induced MPO ANCA and MPO AAV in vivo in rats.
Results When NETs were induced by PMA with PTU using human neutrophils in vitro, abnormal conformation of NETs was observed. Interestingly, the abnormal NETs were hardly digested by DNase I. Moreover, rats immunized with the abnormal NETs, which had been induced by PMA with PTU using rat neutrophils, produced MPO ANCA and developed pulmonary capillaritis. When rats were given oral PTU with intraperitoneal injection of PMA, pauci-immune glomerulonephritis and pulmonary capillaritis occurred with MPO ANCA production in the serum.
Conclusion Our findings indicate that abnormal conformation and impaired degradation of NETs induced by PTU are involved in the pathogenesis of PTU-induced MPO ANCA production and MPO AAV. These findings suggest that disordered NETs can be critically implicated in the pathogenesis of MPO AAV.

Proteasome subunit beta 5t expression in cervical ectopic thymoma
Utano Tomaru, Yosuke Yamada, Akihiro Ishizu, Toru Kuroda, Yoshihiro Matsuno, Masanori Kasahara
JOURNAL OF CLINICAL PATHOLOGY, 65, 9, 858, 859, B M J PUBLISHING GROUP, Sep. 2012, [Peer-reviewed]
English

Decrease of Peripheral and Intestinal NKG2A-Positive T Cells in Patients with Ulcerative Colitis
Takehiko Katsurada, Waka Kobayashi, Utano Tomaru, Tomohisa Baba, Shigeru Furukawa, Akihiro Ishizu, Kazuyoshi Takeda, Naoya Sakamoto, Masahiro Asaka, Hiroshi Takeda, Masanori Kasahara
PLOS ONE, 7, 9, e44113, PUBLIC LIBRARY SCIENCE, Sep. 2012, [Peer-reviewed]
English, Scientific journal, To investigate the role of inhibitory natural killer receptors (iNKRs) in inflammatory bowel disease (IBD), we analyzed the expression of NKG2A, one of the iNKRs, on T cells in a mouse colitis model and human IBD. During the active phase of dextran sulfate sodium (DSS)-induced mouse colitis, the frequency of NKG2A+ T cells was significantly decreased in the peripheral blood, and increased in the intestine, suggesting the mobilization of this T cell subset to the sites of inflammation. Administration of anti-NKG2A antibody increased the number of inflammatory foci in DSS-induced colitis, suggesting the involvement of NKG2A+ T cells in this colitis model. In ulcerative colitis (UC) patients, the frequency of peripheral blood NKG2A+ T cells was significantly decreased, compared with Crohn's disease (CD) patients and healthy controls, regardless of clinical conditions such as treatment modalities and disease activity. Notably, in sharp contrast to the DSS-induced mouse colitis model, the frequency of NKG2A+ cells among intestinal T cells was also decreased in UC patients. These results suggest that inadequate local infiltration of NKG2A+ T cells may be involved in the pathogenesis of UC.

Novel Process of Intrathymic Tumor-Immune Tolerance through CCR2-Mediated Recruitment of Sirp alpha(+) Dendritic Cells: A Murine Model
Tomohisa Baba, Mohamed El Sherif Badr, Utano Tomaru, Akihiro Ishizu, Naofumi Mukaida
PLOS ONE, 7, 7, e41154, PUBLIC LIBRARY SCIENCE, Jul. 2012, [Peer-reviewed]
English, Scientific journal, Immune surveillance system can detect more efficiently secretory tumor-specific antigens, which are superior as a target for cancer immunotherapy. On the contrary, immune tolerance can be induced in the thymus when a tumor antigen is massively secreted into circulation. Thus, the secretion of tumor-specific antigen may have contradictory roles in tumor immunity in a context-dependent manner. However, it remains elusive on the precise cellular mechanism of intrathymic immune tolerance against tumor antigens. We previously demonstrated that a minor thymic conventional dendritic cell (cDC) subset, CD8 alpha(-)Sirp alpha(+) cDCs, but not the major subset, CD8 alpha(+)Sirp alpha(-) cDCs can selectively capture blood-borne antigens and crucially contribute to the self-tolerance. In the present study, we further demonstrated that Sirp alpha(+) cDCs can capture a blood-borne antigen leaking inside the interlobular vascular-rich regions (IVRs). Blood-borne antigen selectively captured by Sirp alpha(+) cDCs can induce antigen-specific Treg generation or negative selection, depending on the immunogenicity of the presented antigen. Furthermore, CCR2 expression by thymic Sirp alpha(+) cDCs and abundant expression of its ligands, particularly, CCL2 by tumor-bearing mice prompted us to examine the function of thymic Sirp alpha(+) cDCs in tumor-bearing mice. Interestingly, tumor-bearing mice deposited CCL2 inside IVRs in the thymus. Moreover, tumor formation induced the accumulation of Sirp alpha(+) cDCs in IVRs under the control of CCR2-CCL2 axis and enhanced their capacity to take up antigens, resulting in the shift from Treg differentiation to negative selection. Finally, intrathymic negative selection similarly ensued in CCR2-competent mice once the tumor-specific antigen was secreted into bloodstream. Thus, we demonstrated that thymic Sirp alpha(+) cDCs crucially contribute to this novel process of intrathymic tumor immune tolerance.

The activity of the inosine triphosphate pyrophosphatase affects toxicity of 6-mercaptopurine during maintenance therapy for acute lymphoblastic leukemia in Japanese children.
Tanaka Y, Manabe A, Nakadate H, Kondoh K, Nakamura K, Koh K, Tomaru U, Kikuchi A, Komiyama T
Leukemia research, 36, 5, 560, 564, May 2012, [Peer-reviewed], [International Magazine]
English, Scientific journal, The association between inosine triphosphate pyrophosphatase (ITPA) activity and toxicity of 6-mercaptopurine (6-MP) was retrospectively evaluated in 65 Japanese children with acute lymphoblastic leukemia (ALL). Patients with an ITPA activity of less than 126 μmol/h/gHb presented with hepatotoxicity more frequently than those with higher ITPA activity (p<0.01). The average 6-MP dose during maintenance therapy administered to two patients with the ITPA deficiency was lower than that given to the other patients. Measuring ITPA activity is important for ensuring the safety of maintenance therapy for Asians with ALL because thiopurine S-methyl transferase mutations are rare in the Asian population.

Expression of proteasome subunit β5t in thymic epithelial tumors
山田 洋介, 外丸 詩野, 石津 明洋, 木内 隆之, 丸川 活司, 松野 吉宏, 笠原 正典
北海道醫學雜誌 = Acta medica Hokkaidonensia, 87, 2, 71, 71, 01 Apr. 2012
Japanese

プロテアソーム機能の低下とCOPDに対する病理作用　　　　　　　　　　　　　　　
山田 洋介, 外丸 詩野, 木内 隆之, 石津 明洋, 松野 吉宏, 笠原 正典
日本病理学会会誌, 101, 1, 279, 279, (一社)日本病理学会, Mar. 2012, [Peer-reviewed]
Japanese

肺腺癌に伴うトルソー症候群により肺高血圧症を合併した一剖検例　　　　　　　　　　　　　　　
今本 鉄平, 大塚 紀幸, 山田 洋介, 外丸 詩野, 高階 太一, 石津 明洋, 笠原 正典
日本病理学会会誌, 101, 1, 439, 439, (一社)日本病理学会, Mar. 2012, [Peer-reviewed]
Japanese

Decreased Proteasomal Activity Causes Age-Related Phenotypes and Promotes the Development of Metabolic Abnormalities
Utano Tomaru, Satomi Takahashi, Akihiro Ishizu, Yukiko Miyatake, Aya Gohda, Sayuri Suzuki, Ayako Ono, Jiro Ohara, Tomohisa Baba, Shigeo Murata, Keiji Tanaka, Masanori Kasahara
AMERICAN JOURNAL OF PATHOLOGY, 180, 3, 963, 972, ELSEVIER SCIENCE INC, Mar. 2012, [Peer-reviewed]
English, Scientific journal, The proteasome is a multicatalytic enzyme complex responsible for the degradation of both normal and damaged proteins. An age-related decline in proteasomal activity has been implicated in various age-related pathologies. The relevance of decreased proteasomal activity to aging and age-related diseases remains unclear, however, because suitable animal models are not available. In the present study, we established a transgenic (Tg) mouse model with decreased proteasomal chymotrypsin-like activity. Tg mice exhibited a shortened life span and developed age-related phenotypes. In Tg mice, polyubiquitinated and oxidized proteins accumulated, and the expression levels of cellular proteins such as Bcl-xL and RNase L were altered. When Tg mice were fed a high-fat diet, they developed more pronounced obesity and hepatic steatosis than did wild-type mice. Consistent with its role in lipid droplet formation, the expression of adipose differentiation-related protein (ADRP) was elevated in the livers of Tg mice. Of note, obesity and hepatic steatosis induced by a high-fat diet were more pronounced in aged than in young wild-type mice, and aged wild-type mice had elevated levels of ADRP, suggesting that the metabolic abnormalities present in Tg mice mimic those in aged mice. Our results proteasomal chymotrypsin-like activity affects longevity and aggravates age-related metabolic disorders, such as obesity and hepatic steatosis. (Am J Pathol(2012, 180:963-972; DOI. 10.1016/j.ajpath.2011.11.012)

Comparative genomic analysis of the proteasome beta 5t subunit gene: implications for the origin and evolution of thymoproteasomes
Yoichi Sutoh, Mizuho Kondo, Yuko Ohta, Tatsuya Ota, Utano Tomaru, Martin F. Flajnik, Masanori Kasahara
IMMUNOGENETICS, 64, 1, 49, 58, SPRINGER, Jan. 2012, [Peer-reviewed]
English, Scientific journal, The thymoproteasome is a recently discovered, specialized form of 20S proteasomes expressed exclusively in the thymic cortex. Although the precise molecular mechanism by which the thymoproteasome exerts its function remains to be elucidated, accumulating evidence indicates that it plays a crucial role in positive selection of T cells. In the present study, we analyzed the evolution of the beta 5t subunit, a beta-type catalytic subunit uniquely present in thymoproteasomes. The gene coding for the beta 5t subunit, designated PSMB11, was identified in the cartilaginous fish, the most divergent group of jawed vertebrates compared to the other jawed vertebrates, but not in jawless vertebrates or invertebrates. Interestingly, teleost fish have two copies of apparently functional PSMB11 genes, designated PSMB11a and PSMB11b, that encode beta 5t subunits with distinct amino acids in the S1 pocket. BLAST searches of genome databases suggest that birds such as chickens, turkey, and zebra finch lost the PSMB11 gene, and have neither thymoproteasomes nor immunoproteasomes. In mammals, reptiles, amphibians, and teleost fishes, the PSMB11 gene (the PSMB11a gene in teleost fish) is located next to the PSMB5 gene coding for the beta 5 subunit of the standard 20S proteasome, indicating that the PSMB11 gene arose by tandem duplication from the evolutionarily more ancient PSMB5 gene. The general absence of introns in PSMB11 and an unusual exon-intron structure of jawed vertebrate PSMB5 suggest that PSMB5 lost introns and duplicated in tandem in a common ancestor of jawed vertebrates, with PSMB5 subsequently gaining two introns and PSMB11 remaining intronless.

Acute Renal Failure due to Thrombotic Microangiopathy in Patient with Scleroderma: Autopsy Case Report.
Ishizu A, Fukaya S, Tomaru U, Katsumata K, Suzuki A, Umemoto Y, Furusaki A, Amasaki Y
Annals of vascular diseases, 5, 4, 458, 461, 2012, [Peer-reviewed]

Abundant neutrophil extracellular traps in thrombus of patient with microscopic polyangiitis
Daigo Nakazawa, Utano Tomaru, Chiho Yamamoto, Satoshi Jodo, Akihiro Ishizu
FRONTIERS IN IMMUNOLOGY, 3, 333, FRONTIERS RESEARCH FOUNDATION, 2012, [Peer-reviewed]
English, Scientific journal, This is a case study of a patient diagnosed with microscopic polyangiitis (MPA) and complicated with deep vein thrombosis (DVT), who died of respiratory failure despite treatment. Autopsy revealed severe crescentic glomerulonephritis and massive alveolar hemorrhage. The thrombus contained abundant neutrophils. Although it is reported that patients with ANCA-associated vasculitis (AAV) have an increased risk of DVT, it remains elusive why they are prone to thrombosis. A recent study has demonstrated the presence of neutrophil extracellular traps (NETs), a newly recognized mode of neutrophil cell-death, in glornerular crescents of MPA patients. Interestingly, NETs were identified in the thrombus as well as in the glomerular crescents in the present case. When compared to other thrombi unrelated to MPA, the amount of NETs was significantly greater in the MPA patient. On the other hand, NETs are critically involved in thrombogenesis because histones within NETs can bind platelets and blood coagulants. Although this is important in regard to containment of microbes within NETs, excessive NETs could cause thrombosis. The collective findings suggest the possibility that thrombosis could be critically associated with MPA via NETs, and that NETs could be a therapeutic target in MPA patients.

プロテアソームサブユニットβ5tの胸腺腫における発現　　　　　　　　　　　　　　　
外丸 詩野, 山田 洋介, 木内 隆之, 丸川 活司, 松野 吉宏, 黒田 徹, 石津 明洋, 笠原 正典
日本病理学会会誌, 100, 2, 37, 37, (一社)日本病理学会, Sep. 2011, [Peer-reviewed]
Japanese

Expression of Proteasome Subunit beta 5t in Thymic Epithelial Tumors
Yosuke Yamada, Utano Tomaru, Akihiro Ishizu, Takayuki Kiuchi, Katsuji Marukawa, Yoshihiro Matsuno, Masanori Kasahara
AMERICAN JOURNAL OF SURGICAL PATHOLOGY, 35, 9, 1296, 1304, LIPPINCOTT WILLIAMS & WILKINS, Sep. 2011, [Peer-reviewed]
English, Scientific journal, Recently, a proteasome beta subunit expressed exclusively in thymic cortical epithelial cells was discovered in mice and humans. This subunit, designated beta 5t, is a component of the thymoproteasome, a specialized type of proteasome implicated in thymic positive selection. To investigate whether beta 5t could serve as a marker for the differential diagnosis of thymic epithelial tumors, we performed immunohistochemical analysis using anti-beta 5t antibody in 54 cases of thymic epithelial tumors comprising 41 cases of thymomas and 13 cases of thymic carcinomas. beta 5t was detected in the neoplastic epithelial cells of thymomas. Among the subtypes of thymoma, expression of beta 5t was observed in most cases of type B thymoma (20 of 21) but not in type A thymomas (0 of 3). In type AB thymomas, beta 5t expression was variable (6 of 17). Type B3 thymomas (4 cases) were positive for beta 5t but negative for CD5, c-kit, and glucose transporter 1 (GLUT-1), which are known as diagnostic markers for thymic carcinomas. In contrast, thymic carcinomas were negative for beta 5t (0 of 13) but expressed at least one and usually all of CD5, c-kit, and GLUT-1. Thus, beta 5t and CD5/c-kit/GLUT-1 were differentially expressed in type B3 thymoma and thymic carcinoma. We tested beta 5t expression in 39 cases of tumors arising from other organs, which showed the specific expression of beta 5t in thymic epithelial tumors. This study demonstrates that beta 5t is expressed in most type B and in some type AB thymomas and is a marker useful in differentiating type B3 thymomas from thymic carcinomas when used in combination with other diagnostic markers.

プロテアソーム機能の低下と喫煙負荷がもたらす老化関連呼吸器系疾患の病態解明　　　　　　　　　　　　　　　
山田 洋介, 外丸 詩野, 木内 隆之, 高橋 里実, 石津 明洋, 松野 吉宏, 笠原 正典
日本病理学会会誌, 100, 1, 332, 332, (一社)日本病理学会, Mar. 2011, [Peer-reviewed]
Japanese

Plasma-Dependent, Antibody- and Fc gamma Receptor-Mediated Translocation of CD8 Molecules from T cells to Monocytes
Sari Iwasaki, Sakiko Masuda, Tomohisa Baba, Utano Tomaru, Kazuaki Katsumata, Masanori Kasahara, Akihiro Ishizu
CYTOMETRY PART A, 79A, 1, 46, 56, WILEY-BLACKWELL, Jan. 2011, [Peer-reviewed]
English, Scientific journal, CD8 alpha beta heterodimers are mainly expressed on cytotoxic T lymphocytes. This study demonstrated the detection of CD8 alpha beta heterodimers on human monocytes by whole blood erythrocyte lysis method in flow cytometry. Results revealed that CD8 alpha beta heterodimers were not produced by monocytes themselves, but were transferred from T cells to monocytes when these cells were coincubated in plasma and with anti-CD8 monoclonal antibody (mAb). For completion of CD8 translocation from T cells to monocytes, cell-to-cell contact between T cells and monocytes, as well as binding of the Fc portion of the anti-CD8 mAb and Fc gamma receptor II (Fc gamma RII) on monocytes were required. Furthermore, the dynamism of cell membrane and cytoskeleton were involved in the mechanism of CD8 translocation. Interestingly, CD3 and alpha beta T cell receptor (TCR) were also transferred from T cells to monocytes accompanied by CD8. These phenomena are consistent with Ab-dependent and Fc gamma R-mediated trogocytosis, which is recently recognized as one of the intercellular communication processes of the immune system. Trogocytosis means exchange of plasma membrane including cell surface molecules in conjugates formed between immune cells. Results of this study could provide another model of trogocytosis and clearly indicated that putative plasma factors were critically implicated in the mechanism of Ab-dependent and Fc gamma R-mediated trogocytosis. (C) 2010 International Society for Advancement of Cytometry

High Prevalence of Multiple Human Papillomavirus Infection in Japanese Patients with Invasive Uterine Cervical Cancer
Hidemichi Watari, Rie Michimata, Motoaki Yasuda, Akihiro Ishizu, Utano Tomaru, Ying Xiong, Mohamed K. Hassan, Noriaki Sakuragi
PATHOBIOLOGY, 78, 4, 220, 226, KARGER, 2011, [Peer-reviewed]
English, Scientific journal, Objective: Multiple human papillomavirus (HPV) infection of the uterine cervix has been suggested as a risk factor for persistent HPV infection, resulting in the development of invasive cervical cancer. The aim of this study was to reveal the actual state of multiple HPV infection in Japanese patients with invasive cervical cancer. Methods: Sixty fresh-frozen invasive cervical cancer tissues were examined for genotyping of HPV. The presence of HPV genotypes was determined with an HPV-DNA array, which can discriminate 25 different HPV genotypes with high sensitivity and specificity. Results: Among 60 samples, 59 (96.7%) were positive for HPV. The three common genotypes were HPV-16 (83.3%), HPV-18 (45.0%) and HPV-52 (28.3%). Multiple HPV infection was observed in 47 of 60 samples (78.3%), among which 42 were infected with more than one high-risk genotype (70.0%). Multiple high-risk HPV infection was significantly more prevalent in patients below 40 years old (14/15, 93.3%) than in patients 40 years of age and over (28/45, 62.2%). Conclusion: The HPV-DNA array is the preferred method to detect HPV genotypes. Multiple HPV infection in Japanese patients with invasive cervical cancer seemed to be more frequent than reported in the literature. Copyright (C) 2011 S. Karger AG, Basel

Frequency and pattern of expression of the stem cell marker CD133 have strong prognostic effect on the surgical outcome of colorectal cancer patients
Shusaku Takahashi, Toshiya Kamiyama, Utano Tomaru, Akihiro Ishizu, Toshiyuki Shida, Mineji Osaka, Yutaka Sato, Yutaka Saji, Michitaka Ozaki, Satoru Todo
ONCOLOGY REPORTS, 24, 5, 1201, 1212, SPANDIDOS PUBL LTD, Nov. 2010, [Peer-reviewed]
English, Scientific journal, CD133 has been reported to be a cancer stem cell marker in colorectal cancer (CRC). The aim of this study was to examine the frequency and pattern of CD133 expression by immunohistochemical methods and evaluate their correlation with clinicopathological features, including patient survival (PS) and recurrence. Tissue specimens of 151 CRC patients who underwent surgical treatment for well-differentiated/moderately differentiated adenocarcinoma and stage I-IV tumors (TNM classification) were immunostained for analyzing CD133 expression. The frequency of CD 133 expression was 91.4% (138/151), and the pattern of expression was divided into membranous and cytoplasmic expression. Of the 151 patients, 136 (90.1%) showed membranous expression, whereas 44 (29.1%) showed cytoplasmic expression. Both expression patterns were seen in 42 (27.8%) patients. The frequency of CD133 overexpression (>50% of stained cells) was 27.2% (41/151); univariate analysis showed CD133 overexpression to be significantly associated with PS, but not recurrence, and multivariate analysis indicated it to be an independent prognostic factor. Multivariate analysis showed membranous overexpression (>50% of stained tumor cells on the membrane), which significantly correlated with histology and chemoresistance of recurrent and stage IV tumors, to be an independent prognostic factor for PS and recurrence. However, multivariate analysis did not indicate cytoplasmic expression, which significantly correlated with histology, lymph node metastasis, TNM stage and lymphatic invasion, as an independent prognostic factor for PS and recurrence. Our results demonstrated that evaluation of the frequency and pattern of CD133 expression is useful for predicting prognosis, recurrence, and chemosensitivity in CRC patients.

Invasive group A streptococcal infection in pregnancy
Takahiro Yamada, Takashi Yamada, Mie K. Yamamura, Kenichi Katabami, Mineji Hayakawa, Utano Tomaru, Shigeki Shimada, Mamoru Morikawa, Toshio Seki, Satoshi Ariga, Kaoru Ishikawa, Tadayoshi Ikebe, Satoshi Gando, Hisanori Minakami
JOURNAL OF INFECTION, 60, 6, 417, 424, W B SAUNDERS CO LTD, Jun. 2010, [Peer-reviewed]
English, We conducted a literature review of 55 pregnancies with symptomatic Group A streptococcus (Streptococcus pyogenes) infection reported in English (20 cases), French (2 cases) and Japanese (33 cases) to seek ways of improving prognosis. Multiparous women (83% [39/47]) in the third trimester (90% [47/52]) were prone to infection from winter to spring (75% [21/28]). Onset was heralded by flu-like symptoms, such as high fever (94% [46/49]), with upper respiratory (40% [22/55]) and/or gastrointestinal symptoms (49% [27/55]). Characteristic findings were early onset of shock (91% [50/55]) and infection-induced strong uterine contraction (73% [40/55]) suggestive of placental abruption. The clinical course was too acute and severe to rescue the mother (58% [32/55] died) and/or infant (66% [39/59] died). However, outcome has improved over the last decade, with rescue of 68% (15/22) of the mothers since 2000, and early use of antibiotics (71% [22/31] survived) and use of intravenous immunoglobulin (91% [10/11] survived) were associated with favourable outcome. Early use of antibiotics and intravenous immunoglobulin may improve outcome of pregnant women suffering from flu-like symptoms, shock and strong uterine contractions suggestive of placental abruption. (C) 2010 The British Infection Society. Published by Elsevier Ltd. All rights reserved.

Cyclic AMP response element-binding protein is implicated in IL-6 production from arthritic synovial cells
Akihiro Ishizu, Asami Abe, Yukiko Miyatake, Tomohisa Baba, Chihiro Iinuma, Utano Tomaru, Takashi Yoshiki
MODERN RHEUMATOLOGY, 20, 2, 134, 138, SPRINGER, Apr. 2010, [Peer-reviewed]
English, Scientific journal, Overproduction of interleukin (IL)-6 from synovial cells is critically involved in the pathogenesis of rheumatoid arthritis (RA). Cyclic adenosine monophosphate (AMP) response element-binding protein (CREB), a leucine zipper transcription factor, is expressed at a high level in synovial cells of patients with RA. Although CREB transactivates IL-6 expression in vascular smooth muscle cells, the relation between CREB expression and IL-6 production from arthritic synovial cells remains unclear. In this study, to determine whether CREB is implicated in IL-6 production from arthritic synovial cells, a dominant negative molecule of activation transcription factor 1 (ATF-1) was transfected into synovial cells obtained from arthritic joints of env-pX rats. These transgenic rats carrying the env-pX gene of human T-cell leukemia virus type-1 develop destructive arthritis with high titers of serum rheumatoid factor and are thus regarded as a suitable model of RA. The dominant negative ATF-1 (ATF-1DN) constitutes a heterodimer with CREB and inhibits CREB function, as CREB/ATF-1DN heterodimers no longer bind to the target sequence of CREB. We showed that transfection of ATF-1DN significantly reduced IL-6 production from arthritic synovial cells. These findings suggest that CREB is implicated in IL-6 production from synovial cells and plays an important role in RA pathogenesis.

プロテアソームサブユニットβ5tの胸腺腫における発現　　　　　　　　　　　　　　　
山田 洋介, 外丸 詩野, 木内 隆之, 丸川 活司, 笠原 正典, 石津 明洋, 松野 吉宏
日本病理学会会誌, 99, 1, 216, 216, (一社)日本病理学会, Mar. 2010, [Peer-reviewed]
Japanese

Human Endogenous Retrovirus-R Env Glycoprotein as Possible Autoantigen in Autoimmune Disease
Naomi Sasaki, Yayoi Ogawa, Chihiro Iinuma, Utano Tomaru, Kazuaki Katsumata, Noriyuki Otsuka, Masanori Kasahara, Takashi Yoshiki, Akihiro Ishizu
AIDS RESEARCH AND HUMAN RETROVIRUSES, 25, 9, 889, 896, MARY ANN LIEBERT INC, Sep. 2009, [Peer-reviewed]
English, Scientific journal, It has long been discussed whether endogenous retroviruses (ERVs) are involved in the pathogenesis of autoimmune diseases. Among various human endogenous retroviruses (HERVs), we have focused on HERV-R. To investigate the biological roles of HERV-R, we earlier established transgenic rats carrying the full sequence of the viral genome. In these HERV-R rats, however, no disease occurred. Another trigger that induces autoimmunity may be essential for the recognition of HERV-R products by the immune system. Thus, in this study, we mated HERV-R rats with env-pX rats (transgenic rats carrying the env-pX gene of human T cell leukemia virus type I) that develop autoimmune diseases, and generated double transgenic (DTG)rats. In DTG rats, autoimmune diseases occurred similarly in env-pX rats. Interestingly, deposition of rat IgM but not IgG was observed on the glomerular endothelial cells. Such IgM deposition was never seen in the parental HERV-R or env-pX rats. We considered that in situ formation of immune complexes consisted of the HERV-R env glycoprotein and anti-HERV-R env IgM antibodies (Abs) in DTG rats, according to the following evidence: (1) No dense deposit, representing deposition of circulating immune complexes, was seen on glomerular endothelial cells. (2) IgM Abs reactive with HERV-R env glycoprotein were generated in the serum. (3) HERV-R env glycoprotein was expressed in the kidney, specifically on glomerular endothelial cells. (4) IgM deposition was partly colocalized with the HERV-R env glycoprotein on the glomeruli. These findings strongly suggest that the HERV-R env glycoprotein is recognized as an autoantigen in the host with autoimmune diseases.

Exclusive expression of proteasome subunit beta 5t in the human thymic cortex
Utano Tomaru, Akihiro Ishizu, Shigeo Murata, Yukiko Miyatake, Sayuri Suzuki, Satomi Takahashi, Taku Kazamaki, Jiro Ohara, Tomohisa Baba, Sari Iwasaki, Kazunori Fugo, Noriyuki Otsuka, Keiji Tanaka, Masanori Kasahara
BLOOD, 113, 21, 5186, 5191, AMER SOC HEMATOLOGY, May 2009, [Peer-reviewed]
English, Scientific journal, The ubiquitin-proteasome pathway, which degrades intracellular proteins, is involved in numerous cellular processes, including the supply of immunocompetent peptides to the antigen presenting machinery. Proteolysis by proteasomes is conducted by three beta subunits, beta 1, beta 2, and beta 5, of the 20S proteasome. Recently, a novel beta subunit expressed exclusively in cortical thymic epithelial cells was discovered in mice. This subunit, designated beta 5t, is a component of the thymoproteasome, a specialized type of proteasomes implicated in thymic positive selection. In this study, we show that, like its mouse counterpart, human beta 5t is expressed exclusively in the thymic cortex. Human beta 5t was expressed in approximately 80% of cortical thymic epithelial cells and some cortical dendritic cells. Human beta 5t was incorporated into proteasomes with two other catalytically active beta subunits beta 1i and beta 2i, forming 20S proteasomes with subunit compositions characteristic of thymoproteasomes. The present study demonstrates, for the first time, the existence of thymoproteasomes in the human thymic cortex, indicating that thymoproteasome function is likely conserved between humans and mice. (Blood. 2009; 113: 5186-5191)

血管内皮細胞における硬化剤の作用に関する研究:最終報告　　　　　　　　　　　　　　　
William Mol, 長尾 宗朝, 斉藤 亮, 斉藤 典子, 林 利彦, 畠 真也, 古川 洋志, 佐々木 了, 山本 有平, 外丸 詩野
日本形成外科学会会誌, 28, 5, 348, 348, (一社)日本形成外科学会, May 2008
Japanese

創傷治癒に関わる新見解 ケロイド体質とCirculating fibrocyte　　　　　　　　　　　　　　　
長尾 宗朝, 外丸 詩野, 石津 明洋, 岩崎 沙理, 馬場 智久, 笠原 正典, 小山 明彦, 小浦場 祥夫, 古川 洋志, 山本 有平
日本病理学会会誌, 97, 1, 201, 201, (一社)日本病理学会, Mar. 2008
Japanese

Rat CD4(+)CD8(+) macrophages kill tumor cells through an NKG2D-and granzyme/perforin-dependent mechanism
Tomohisa Baba, Sari Iwasaki, Takako Maruoka, Akira Suzuki, Utano Tomaru, Hitoshi Ikeda, Takashi Yoshiki, Masanori Kasahara, Akihiro Ishizu
JOURNAL OF IMMUNOLOGY, 180, 5, 2999, 3006, AMER ASSOC IMMUNOLOGISTS, Mar. 2008, [Peer-reviewed]
English, Scientific journal, We previously identified a subpopulation of monocyte/macrophage lineage cells expressing both CD4 and CD8. This subpopulation was expanded in rat peripheral blood and spleen after immunization with adjuvants containing killed tuberculosis germs. CD4(+)CD8(+) monocytes/macrophages obtained from preimmunized rats exhibited a Th1-type cytokine/chemokine profile, expressed high levels of Fas ligand, perforin, granzyme B, and NKR-P2 (rat ortholog of human NKG2D), and killed certain tumor cells. In the present study, we confirmed that CD4(+)CD8(+) monocytes/macrophages are distinct from splenic dendritic cells (DCs) or IFN-producing killer DCs. In vitro cytotoxic assays revealed that CD4(+)CD8(+) macrophages killed tumor cells in a cell-cell contact-dependent manner and that expression of the retinoic acid early transcript 1 (a ligand for NKG2D) made tumor cells susceptible to killing by CD4(+)CD8(+) macrophages. Furthermore, inhibitors of granzyme and perforin significantly decreased cytotoxic activities of CD4(+)CD8(+) macrophages. Consistent with these in vitro findings, preimmunization with adjuvants containing killed tuberculosis germs elevated the expression of granzyme B in tumor-infiltrating CD4(+)CD8(+) macrophages and significantly inhibited the growth of inoculated tumor cells. Our current work demonstrates that CD4(+)CD8(+) macrophages are a unique subpopulation of monocyte/macrophage lineage cells that kill tumor cells in an NKG2D- and granzyme/perforin-dependent mechanism.

Two novel NKG2D ligands of the mouse H60 family with differential expression patterns and binding affinities to NKG2D
Akio Takada, Shigeru Yoshida, Mizuho Kajikawa, Yukiko Miyatake, Utano Tomaru, Masaharu Sakai, Hitoshi Chiba, Katsumi Maenaka, Daisuke Kohda, Kazunori Fugo, Masanori Kasahara
JOURNAL OF IMMUNOLOGY, 180, 3, 1678, 1685, AMER ASSOC IMMUNOLOGISTS, Feb. 2008, [Peer-reviewed]
English, Scientific journal, H60, originally described as a dominant minor histocompatibility Ag, is an MHC class I-like molecule that serves as a ligand for the NKG2D receptor. In the present study, we identified two novel mouse chromosome 10-encoded NKG2D ligands structurally resembling H60. These ligands, which we named H60b and H60c, encode MHC class I-like molecules with two extracellular domains. Whereas H60b has a transmembrane region, H60c is a GPI-anchored protein. Recombinant soluble H60b and H60c proteins bound to NKG2D with affinities typical of cell-cell recognition receptors (K(d) = 310 nM for H60b and K(d) = 8.7 mu M for H60c). Furthermore, expression of H60b or H60c rendered Ba/F3 cells susceptible to lysis by NK cells, thereby establishing H60b and H60c as functional ligands for NKG2D. H60b and H60c transcripts were detected only at low levels in tissues of healthy adult mice. Whereas H60b transcripts were detectable in various tissues, H60c transcripts were detected mainly in the skin. Infection of mouse embryonic fibroblasts with murine cytomegalovirus induced expression of H60b, but not H60c or the previously known H60 gene, indicating that transcriptional activation of the three types of H60 genes is differentially regulated. The present study adds two new members to the current list of NKG2D ligands.

Evaluation of the sclerotherapeutic efficacy of ethanol, polidocanol, and OK-432 using an in vitro model
William Mol, Hiroshi Furukawa, Satoru Sasaki, Utano Tomaru, Toshihiko Hayashi, Akira Saito, Munetomo Nagao, Noriko Saito, Shinya Hata, Yuhei Yamamoto
DERMATOLOGIC SURGERY, 33, 12, 1452, 1459, BLACKWELL PUBLISHING, Dec. 2007, [Peer-reviewed]
English, Scientific journal, BACKGROUND Sclerosants are used to treat vascular malformations. Owing to variations in the flow, the injected concentrations and the duration of exposure of these sclerosants are altered. Therefore, the clinical effectiveness of sclerotherapy is variable.
OBJECTIVE The objective was to evaluate the differences in clinical response, usually observed among ethanol, polidocanol, and OK-432, using an in vitro sclerotherapy model.
METHODS Endothelial cells were cultured and exposed to different concentrations of the sclerosants for 5 seconds and the remaining viable cells were counted using a MTT assay kit. Dyes were used to visualize the morphologic changes. Precipitant formation in blood was also evaluated. Finally, the degree of ICAM-1 expression, after exposure to lower concentrations of these sclerosants, was studied using immunocytochemistry.
RESULTS Only ethanol causes precipitant formation and kills almost all cells from 30% concentration. Polidocanol begins to disrupt the cell membrane from 0.0125% onward. Only OK-432 induces ICAM-1 expression.
CONCLUSION Ethanol's strong precipitant-forming effect may induce thromboembolism, thus enhancing sclerosis. Polidocanol's endothelial cell-lysing effect was clearly documented. OK-432 may mediate its effect by inducing inflammatory response of the endothelium via ICAM-1 expression. This in vitro model may be useful in evaluating other sclerosants as well.

Enhanced production of p24 Gag protein in HIV-1-infected rat cells fused with uninfected human cells
Jing Chen, Xudong Zhao, Yurong Lai, Akira Suzuki, Utano Tomaru, Akihiro Ishizu, Akio Takada, Hitoshi Ikeda, Masanori Kasahara, Takashi Yoshiki
EXPERIMENTAL AND MOLECULAR PATHOLOGY, 83, 1, 125, 130, ACADEMIC PRESS INC ELSEVIER SCIENCE, Aug. 2007, [Peer-reviewed]
English, Scientific journal, Although many human molecules have been suggested to affect replication of human immunodeficiency virus type 1 (HIV-1), the distribution of such cofactors in human cell types is not well understood. Rat W31/D4R4 fibroblasts expressing human CD4 and CXCR4 receptors were infected with HIV-1. The provirus was integrated in the host genome, but only a limited amount of p24 Gag protein was produced in the cells and culture supernatants. Here we found that p24 production was significantly increased by fusing HIV-1-infected W31/D4R4 cells with uninfected human cell lines of T-cell, B-cell, or macrophage lineages. These findings suggest that human cellular factors supporting HIV-1 replication are distributed widely in cells of lymphocyte and macrophage lineages. We also examined whether the amount of p24 produced by rat-human hybrid cells was correlated with expression levels of specific human genes. The results suggested that HP68 and MHC class II transactivator (CIITA) might up- and down-regulate p24 production, respectively. It was also suggested that HIV-1 replication is affected by molecules other than those examined in this study, namely, cyclin T1, cyclin-dependent kinase 9, CRM1, HP68, and CIITA. (c) 2006 Elsevier Inc. All rights reserved.

MHC class I-like MILL molecules are beta(2)-microglobulin-associated, GPI-anchored glycoproteins that do not require TAP for cell surface expression
Mizuho Kajikawa, Tomohisa Baba, Utano Tomaru, Yutaka Watanabe, Satoru Koganei, Sachiyo Tsuji-Kawahara, Naoki Matsumoto, Kazuo Yamamoto, Masaaki Miyazawa, Katsumi Maenaka, Akihiro Shizu, Masanori Kasahara
JOURNAL OF IMMUNOLOGY, 177, 5, 3108, 3115, AMER ASSOC IMMUNOLOGISTS, Sep. 2006, [Peer-reviewed]
English, Scientific journal, MILL (MHC class I-like located near the leukocyte receptor complex) is a family of MHC class I-like molecules encoded outside the MHC, which displays the highest sequence similarity to human MICA/B molecules among known class I molecules. In the present study, we show that the two members of the mouse MILL family, MILL1 and MILL2, are GPI-anchored glycoproteins associated with beta(2)-microglobulin (beta(2)m) and that cell surface expression of MILL1 or MILL2 does not require functional TAP molecules. MILL1 and MILL2 molecules expressed in bacteria could be refolded in the presence of beta(2)m, without adding any peptides. Hence, neither MILL1 nor MILL2 is likely to be involved in the presentation of peptides. Immunohistochemical analysis revealed that MILL1 is expressed in a subpopulation of thymic medullary epithelial cells and a restricted region of inner root sheaths in hair follicles. The present study provides additional evidence that MILL is a class I family distinct from MICA/B.

Role of neuronal interferon-gamma in the development of myelopathy in rats infected with human T-cell leukemia virus type 1
Y Miyatake, H Ikeda, A Ishizu, T Baba, T Ichihashi, A Suzuki, U Tomaru, M Kasahara, T Yoshiki
AMERICAN JOURNAL OF PATHOLOGY, 169, 1, 189, 199, AMER SOC INVESTIGATIVE PATHOLOGY, INC, Jul. 2006, [Peer-reviewed]
English, Scientific journal, Human T-cell leukemia virus type I (HTLV-1) is the causative agent of not only adult T-cell leukemia but also HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Among die rat strains infected with HTLV-1, chronic progressive myelopathy, named HAM rat disease, occurs exclusively in WKAH rats. In the present study, we found that HTLV-1 infection induces interferon (IFN)-gamma production in the spinal cords of HAM-resistant strains but not in those of WKAH rats. Neurons were the major cells that produced IFN-gamma in HTLV-1-infected, HAM-resistant strains. Administration of IFN-gamma suppressed expression of pX, the gene critically involved in the onset of HAM rat disease, in an HTLV-1-immortalized rat T-cell line, indicating that IFN-gamma protects against the development of HAM rat disease. The inability of WKAH spinal cord neurons to produce IFN-gamma after infection appeared to stem from defects in signaling through the interleukin (IL)-12 receptor. Specifically, WKAH-derived spinal cord cells were unable to up-regulate the IL-12 receptor beta 2 gene in response to IL-12 stimulation. We suggest that the failure of spinal cord neurons to produce IFN-gamma through the IL-12 pathway is involved in the development of HAM rat disease.

CD4(+)/CD8(+) macrophages infiltrating at inflammatory sites: a population of monocytes/macrophages with a cytotoxic phenotype
T Baba, A Ishizu, S Iwasaki, A Suzuki, U Tomaru, H Ikeda, T Yoshiki, M Kasahara
BLOOD, 107, 5, 2004, 2012, AMER SOC HEMATOLOGY, Mar. 2006, [Peer-reviewed]
English, Scientific journal, We found a population of nonlymphoid cells expressing both CD4 and CD8 in peripheral blood mononuclear cells (PBMCs) of human T-cell leukemia virus type-I pX transgenic rats with autoimmune diseases. These cells, which showed a monocytic phenotype, were also found in wild-type rats, and their number increased by adjuvant-assisted immunization. GM-CSF increased the number of these double-positive (DIP) monocytes in PBMCs. Consistent with the idea that DIP monocytes differentiate into DIP macrophages at sites of inflammation, we found infiltration of DIP macrophages at the site of myosin-induced myocarditis in wild-type rats; these cells exhibited a T-helper 1 (Th1)-type cytokine/chemokine profile and expressed high levels of Fas ligand, perforin, granzyme B, and NKR-P2 (rat orthologue of human NKG2D). Adoptive transfer of GFP-positive spleen cells confirmed hematogenous origin of DIP macrophages. DIP monocytes had a cytotoxic phenotype similar to DID macrophages, indicating that this phenotypic specialization occurred before entry into a tissue. In line with this, DIP monocytes killed tumor cells in vitro. Combined evidence indicates that certain inflammatory stimuli that induce GM-CSF trigger the expansion of a population of DIP monocytes with a cytotoxic phenotype and that these cells differentiate into macrophages at inflammatory sites. Interestingly, human PBMCs also contain DIP monocytes.

HLA-DRB5*0101 and-DRB1*1501 expression in the multiple sclerosis-associated HLA-DR15 haplotype
E Prat, U Tomaru, L Sabater, DM Park, R Granger, N Kruse, JM Ohayon, MP Bettinotti, R Martin
JOURNAL OF NEUROIMMUNOLOGY, 167, 1-2, 108, 119, ELSEVIER SCIENCE BV, Oct. 2005, [Peer-reviewed]
English, Scientific journal, The HLA region, and particularly the DR15 haplotype (containing the two DRB* genes DRB1*1501 and DRB5*0101 and the tightly linked DQ alleles DQA*0102 and DQB1*0602, which together form the DQw6 molecule) in Caucasians, shows the strongest genetic association with multiple sclerosis (MS). In the DR15 haplotype, two beta-chains HLA-DRB1*1501 and -DRB5*0101 are co-expressed resulting in two different surface HLA-DR alpha beta heterodimers, DR2b and DR2a. Most previous studies focused on DRB1*1501, however, both DR2a, and DR2b may contribute to MS pathogenesis via antigen presentation to myelin-specific T lymphocytes. We therefore analyzed the expression of the two DR15 genes in various antigen presenting cells (APCs), central nervous system and thymic tissues. Transcript levels were higher for DRB5*0101 in all cell types and tissues. Both HLA-DR heterodimers were expressed at significant levels on the cell surface, where they showed a differential expression pattern in different APCs. They were similarly regulated after stimulation with interferon-gamma and interieukin-4. Finally, immunohistochemistry experiments indicated that both molecules were expressed in thymic tissue. Our results encourage future research to investigate the potential functional relevance of both genes for the pathogenesis of MS. (C) 2005 Elsevier B.V. All rights reserved.

Aberrant gene expression by CD25(+)CD4(+) immunoregulatory T cells in autoimmune-prone rats carrying the human T cell leukemia virus type-I gene
H Hayase, A Ishizu, H Ikeda, Y Miyatake, T Baba, M Higuchi, A Abe, U Tomaru, T Yoshiki
INTERNATIONAL IMMUNOLOGY, 17, 6, 677, 684, OXFORD UNIV PRESS, Jun. 2005, [Peer-reviewed]
English, Scientific journal, Transgenic rats expressing the env-pX gene of human T cell leukemia virus type-I under the control of the viral long terminal repeat promoter (env-pX rats) developed systemic autoimmune diseases. Prior to disease manifestation, the immunosuppressive function of CD25(+)CD4(+) T (T-reg) cells was impaired in these rats. Since T cell differentiation appeared to be disordered in env-pX rats, we assumed that the impairment of T-reg cells might be caused by an abortive differentiation in the thymus. However, reciprocal bone marrow transfers between env-pX and wild-type rats revealed that direct effects of the transgene unrelated to the thymus framework induced the abnormality of T-reg cells. To identify molecular changes, comparative analyses were done between env-pX and wild-type T-reg cells. Expression of the Foxp3 gene and cell-surface markers supported a naive phenotype for env-pX T-reg cells. Array analyses of gene expression showed some interesting profiles, e.g. up-regulation of genes associated with the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathways in env-pX T-reg cells. Additionally, expression of the suppressor of cytokine signaling (SOCS) family genes, which inhibit the JAK/STAT signals, was extremely low in env-pX T-reg cells. These findings suggest that the transgene may mediate the down-regulation of the SOCS family genes and that subsequent excess signals through the JAK/STAT pathways may result in the loss of function of env-pX T-reg cells. We suggest that investigation of the pathology of T-reg cells in our autoimmune-prone rat model may aid in understanding the roles of T-reg cells in human autoimmune diseases.

Chronic graft-versus-host disease-like autoimmune disorders spontaneously occurred in rats with neonatal thymus atrophy
T Baba, A Ishizu, H Ikeda, Y Miyatake, T Tsuji, A Suzuki, U Tomaru, T Yoshiki
EUROPEAN JOURNAL OF IMMUNOLOGY, 35, 6, 1731, 1740, WILEY-V C H VERLAG GMBH, Jun. 2005, [Peer-reviewed]
English, Scientific journal, We earlier reported that the human T cell leukemia virus type-1 pX gene transduced into rat thymic epithelial cells had an impact on biology of the cells. We report here that FW-pX rats born by mating of F344 transgenic rats expressing the pX gene without tissue specificity with nontransgenic Wistar rats developed disorders, including atrophy of the thymus, lymphocytopenia, and inflammatory cell infiltration into multiple organs, similar to events in chronic graft-vs.-host disease (GVHD). Vanishment of thymic epithelial cells especially in the cortex and marked depletion of CD4 CD8 double-positive thymocytes were evident in the neonatal thymus in these rats. The relative abundance of CD8 compared to CD4 T cells may be related to dominant infiltration of CD8 T cells into the affected organs. Additionally, adoptive transfer of FW-pX splenocytes could induce lymphocytic infiltration into sublethally irradiated wild-type syngeneic recipients. Analysis of the expression level of the Foxp3 gene in peripheral blood mononuclear cells revealed that the numbers of immunoregulatory T cells were less in FW-pX rats than in wild-type rats. The collective evidence suggested that the FW-pX rats spontaneously developed chronic GVHD-like autoimmune diseases, following abortive differentiation of T cells in the thymus in early days of the newborn. This rat model may shed light on the pathogenesis of chronic GVHD and also other systemic autoimmune diseases, the etiology of which is unknown.

Virus-induced dysfunction of CD4+CD25+T cells in patients with HTLV-I-associated neuroimmunological disease
Y Yamano, N Takenouchi, HC Li, U Tomaru, K Yao, CW Grant, DA Maric, S Jacobson
JOURNAL OF CLINICAL INVESTIGATION, 115, 5, 1361, 1368, AMER SOC CLINICAL INVESTIGATION INC, May 2005, [Peer-reviewed]
English, Scientific journal, CD4(+)CD25(+) Tregs are important in the maintenance of immunological self tolerance and in the prevention of autoimmune diseases. As the CD4(+)CD25(+) T cell population in patients with human T cell lymphotropic virus type I-associated (HTLV-I-associated) myelopathy/tropical spastic paraparesis (HAM/TSP) has been shown to be a major reservoir for this virus, it was of interest to determine whether the frequency and function of CD4(+)CD25(+) Tregs in HAM/TSP patients might be affected. in these cells, both mRNA and protein expression of the forkhead transcription factor Foxp3, a specific marker of Tregs, were lower than those in CD4(+)CD25(+) T cells from healthy individuals. The virus-encoded transactivating HTLV-1 tax gene was demonstrated to have a direct inhibitory effect on Foxp3 expression and function of CD4(+)CD25(+) T cells. This is the first report to our knowledge demonstrating the role of a specific viral gene product (HTLV-I Tax) on the expression of genes associated with Tregs (in particular, foxp3) resulting in inhibition of Treg function. These results suggest that direct human retroviral infection of CD4(+)CD25(+) T cells may be associated with the pathogenesis of HTLV-I-associated neurologic disease.

Peptide/HLA-GFP complexes: Detection of antigen-specific T cells by acquisition of peptide/HLA-GFP complexes
Utano Tomaru, Yoshihisa Yamano, Steven Jacobson
Analyzing T Cell Responses: How to Analyze Cellular Immune Responses against Tumor Associated Antigens, 261, 274, Springer Netherlands, 2005, [Peer-reviewed]
English, In book, Antigen-specific T cell responses are essential in host immune defense in health and disease. For many years, it was believed that the immune system was effective only in combating infectious diseases caused by invading agents such as bacteria and viruses. More recently, however, the immune system has been shown to play a central role in protection and recovery against cancer. This latter role is not well understood, but there are numerous reports that the immune system slows down the growth and spread of tumors in cancer patients. Many clinical trials for cancer immunotherapy are in progress and use anti-tumor vaccinations that have been designed to elicit antigen-specific T cell responses. Therefore, in defining anti-tumor immune responses that may be used for immunotherapy trials, the detection and quantitative analysis of antigen-specific T cell populations has been an important step toward understanding the cellular immune response in patients. In this chapter, we introduce a newly established system for the analysis of antigen-specific T cells, which is based on the technology using artificial antigen presenting cell expressing human leukocyte antigen (HLA)-A*201 coupled to the enhanced green fluorescent protein (GFP) (HmyA2GFP cells). Antigen-specific CD8+ T cells have been demonstrated to acquire peptide-major histocompatibility complex (MHC) clusters through T cell receptor-mediated endocytosis upon specific antigen stimulation. We generated an antigen-presenting cell expressing HLA-A*201 coupled to the GFP which when pulsed with antigenic peptide will deliver the GFP to an antigen-specific T cell. We demonstrated the quantitative identification of human T-cell lymphotropic virus type I (HTLV-I) Tax11-19 peptide-specific T cell populations in peripheral blood mononuclear cells from patients with HTLV-I associated neurologic disease and defined a novel CD8+ T cell epitope in the HTLV-I Envelope region. © 2005 Springer.

Increased expression of human T lymphocyte virus type I (HTLV-I) Tax11-19 peptide-human histocompatibility leukocyte antigen A*201 complexes on CD4(+) CD25(+) T cells detected by peptide-specific, major histocompatibility complex-restricted antibodies in patients with HTLV-I-associated neurologic disease
Y Yamano, CJ Cohen, N Takenouchi, K Yao, U Tomaru, HC Li, Y Reiter, S Jacobson
JOURNAL OF EXPERIMENTAL MEDICINE, 199, 10, 1367, 1377, ROCKEFELLER UNIV PRESS, May 2004, [Peer-reviewed]
English, Scientific journal, Human T lymphocyte virus type I (HTLV-I)-associated chronic inflammatory neurological disease (HTLV-I-associated myelopathy/tropical spastic paraparesis [HAM/TSP]) is suggested to be an immunopathologically mediated disorder characterized by large numbers of HTLV-I Tax-specific CD8(+) T cells. The frequency of these cells in the peripheral blood and cerebrospinal fluid is proportional to the amount of HTLV-I proviral load and the levels of HTLV-I tax mRNA expression. As the stimulus for these virus-specific T cells are immunodominant peptide-human histocompatibility leukocyte antigen (HLA) complexes expressed on antigen-presenting cells, it was of interest to determine which cells express these complexes and at what frequency. However, until now, it has not been possible to identify and/or quantify these peptide-HLA complexes. Using a recently developed antibody that specifically recognizes Tax11-19 peptide-HLA-A(star)201 complexes, the level of Tax11-19-HLA-A(star)201 expression on T cells was demonstrated to be increased in HAM/TSP and correlated with HTLV-I proviral DNA load, HTLV-I tax mRNA load, and HTLV-I Tax-specific CD8(+) T cell frequencies. Furthermore, CD4(+) CD25(+) T cells were demonstrated to be the major reservoir of HTLV-I provirus as well as Tax11-19 peptide-HLA-A*201 complexes. These results indicate that the increased detection and visualization of peptide-HLA complexes in HAM/TSP CD4(+) CD25(+) T cell subsets that are shown to stimulate and expand HTLV-I Tax-specific CD8(+) T cells may play an important role in the pathogenesis of HTLV-I-associated neurological disease.

Provirus expansion and deregulation of apoptosis-related genes in the spinal cord of a rat model for human T-lymphocyte virus type I-associated myeloneuropathy
U Tomaru, H Ikeda, XY Jiang, O Ohya, T Yoshiki
JOURNAL OF NEUROVIROLOGY, 9, 5, 530, 538, TAYLOR & FRANCIS INC, Oct. 2003, [Peer-reviewed]
English, Scientific journal, Apoptosis of the spinal oligodendrocytes is the main factor linked to the pathogenesis of human T-lymphocyte virus type I ( HTLV-I)-induced myeloneuropathy in rats (HAM rat). To clarify apoptosis-related mechanisms, expression of apoptosis-related genes in the spinal cord of these rats was chronologically examined by means of a semiquantitative reverse transcriptase-polymerase chain reaction. Provirus expansion and increment of HTLV-I pX mRNA were evident at 7 months after the induced infection. Tumor necrosis factor-alpha increased gradually soon after pX expression. The expression of a major apoptosis-resistant gene, bcl-2, was markedly suppressed at a period of the provirus expansion and bax was also down-regulated. p53 was consistently expressed at high levels. These findings were never observed in spinal cords of HAM-resistant strains with HTLV-I infection even throughout their entire life. Collective evidence suggests that the local provirus expansion and deregulation of apoptosis-related genes, especially down-regulation of bcl-2, may lead to apoptosis of oligodendrocytes, thus being a major pathogenetic pathway in the HTLV-I-induced myeloneuropathy.

Detection of virus-specific T cells and CD8(+) T-cell epitopes by acquisition of peptide-HLA-GFP complexes: analysis of T-cell phenotype and function in chronic viral infections
U Tomaru, Y Yamano, M Nagai, D Maric, PTP Kaumaya, W Biddison, S Jacobson
NATURE MEDICINE, 9, 4, 469, 475, NATURE PUBLISHING GROUP, Apr. 2003, [Peer-reviewed]
English, Scientific journal, Antigen-specific CD8(+) T cells acquire peptide-major histocompatibility complex (MHC) clusters through T-cell receptor (TCR)-mediated endocytosis after specific antigen stimulation. We generated an antigen-presenting cell (APC) expressing human leukocyte antigen (HLA)-A*201 coupled to the enhanced green fluorescent protein (GFP), which delivered GFP to an antigen-specific T cell when pulsed with antigenic peptide. We quantitatively identified human T-cell lymphotropic virus type I ( HTLV-I) Tax(11-19) peptide-specific T-cell populations in peripheral blood mononuclear cells (PBMCs) from patients with HTLV-I-associated neurologic disease and defined a new CD8(+) T-cell epitope in the HTLV-I envelope region. Acquisition of peptide-HLA-GFP complexes by antigen-specific T cells could distinguish, with respect to phenotype and perforin production, T cells from the chronic viral infections cytomegalovirus and HTLV-I. This approach will be a powerful tool in understanding the role of antigen-specific T-cell responses in health and disease.

Direct phenotypic analysis of human MHC class I antigen presentation: Visualization, quanititation, and in situ detection of human viral epitopes using peptide-specific, MHC-restricted human recombinant antibodies
CJ Cohen, O Sarig, Y Yamano, U Tomaru, S Jacobson, Y Reiter
JOURNAL OF IMMUNOLOGY, 170, 8, 4349, 4361, AMER ASSOC IMMUNOLOGISTS, Apr. 2003, [Peer-reviewed]
English, Scientific journal, The advent in recent years of the application of tetrameric arrays of class I peptide-MHC complexes now enables us to detect and study rare populations of Ag-specific CD8(+) T cells. However, aviailable methods cannot visualize or determine the number and distribution of these TCR ligands on individual cells nor detect APCs in tissues. In this study, we describe for the first time studies of human class I peptide-MHC ligand presentation. These studies were facilitated by applying novel tools in the form of peptide-specific, HLA-A2-restricted human recombinant Abs directed toward a viral epitope derived from human T cell lymphotropic virus type I. Using a large human Ab phage display library, we isolated a large panel of recombinant Fab Abs that are specific for a particular peptide-MHC class I complex in a peptide-dependent, MHC-restricted manner. We used these Abs to visualize the specific complex on APCs and virus-infected cells by flow cytometry, to quantify the number of, and visualize in situ, a particular complex on the surface of APCs bearing complexes formed by naturally occurring active intracellular processing of the cognate viral Ag. These findings demonstrate our ability to transform the unique fine specificity, but low intrinsic affinity of TCRs into high affinity soluble Ab molecules endowed with a TCR-like specificity toward human viral epitopes. These molecules may prove to be crucial useful tools for studying MHC class I Ag presentation in health and disease as well as for therapeutic purposes in cancer, infectious diseases, and autoimmune disorders.

Role of HTLV-I specific CTL in HAM/TSP
Y Yamano, U Tomaru, S Jacobson
TWO DECADES OF ADULT T-CELL LEUKEMIA AND HTLV-I RESEARCH, 50, 171, 182, JAPAN SCIENTIFIC SOC PRESS, 2003, [Peer-reviewed]
English, International conference proceedings, Human T-cell lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a chronic, progressive neurological disease characterized by marked degeneration of the spinal cord and the presence of infiltrating T cells and macrophages. HAM/TSP patients harbor very high frequencies of HTLV-I infected T cells and HTLV-I specific CD8+ T cells in both peripheral blood and cerebrospinal fluid. The use of novel molecular and immunological methods has improved our understanding of the underlying pathological mechanisms operative in HAM/TSP. Information presented in this review suggests that HTLV-I infected T cells and HTLV-I specific CD8+ T cells accumulate in central nervous system (CNS), in which high cellular immune response continuously driven by this virus may contribute to the inflammatory process within CNS lesions in HAM/TSP patients.

Further evidence of hepatic transdifferentiation in hepatoid adenocarcinomas of the stomach: quantitative analysis of mRNA for albumin and hepatocyte nuclear factor-4 alpha
T Yano, T Kishimoto, U Tomaru, Y Kawarada, H Kato, T Yoshiki, H Ishikura
PATHOLOGY, 35, 1, 75, 78, CARFAX PUBLISHING, Jan. 2003, [Peer-reviewed]
English, Scientific journal, Aims: To assess the production of a liver-specific protein, albumin, and of a master transcriptional factor, hepatocyte nuclear factor (HNF)-4alpha, in hepatoid adenocarcinoma tissue.
Methods: Standard and quantitative RT-PCR, using five cases of hepatoid and three cases of non-hepatoid gastric adenocarcinoma.
Results: Hepatoid adenocarcinomas expressed similarly large amounts of albumin mRNA as those expressed in hepatocellular carcinoma and normal liver tissues. The observed amounts were several hundred times more than those in non-hepatoid adenocarcinoma and normal stomach tissues. HNF-4alpha mRNA was expressed in all stomach samples examined, and the levels of expression did not quantitatively differ between hepatoid and non-hepatoid adenocarcinomas of the stomach.
Conclusions: These results provide further support of a relationship between hepatic transdifferentiation in hepatoid adenocarcinomas and albumin mRNA expression. Furthermore, transdifferentiation to the hepatocytic phenotype in hepatoid adenocarcinoma tissue was not directly associated with HNF-4alpha expression, thus suggesting that transdifferentiation proceeds by a complicated mechanism.

Correlation of human T-cell lymphotropic virus type 1 (HTLV-1) mRNA with proviral DNA load, virus-specific CD8(+) T cells, and disease severity in HTLV-1-associated myelopathy (HAM/TSP)
Y Yamano, M Nagai, M Brennan, CA Mora, SS Soldan, U Tomaru, N Takenouchi, S Izumo, M Osame, S Jacobson
BLOOD, 99, 1, 88, 94, AMER SOC HEMATOLOGY, Jan. 2002, [Peer-reviewed]
English, Scientific journal, To investigate the role of viral expression in individuals infected with human T-cell lymphotropic virus type 1 (HTLV-1), a real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) of HTLV-1 tax messenger RNA (mRNA) using ABI Prism 7700 Sequence Detection System was developed. Using this system, the HTLV-1 tax mRNA load was compared with HTLV-1 proviral DNA load, HTLV-1 Tax protein expression, HTLV-1 Tax-specific CD8(+) T-cell frequency, and disease severity of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). This approach was a sensitive and specific technique for the precise quantification of HTLV-1 tax mRNA. The total amount of HTLV-1 tax mRNA and mRNA expression level in HTLV-1-infected cells (mRNA/DNA ratio) were higher in HAM/TSP patients than in asymptomatic HTLV-1 carriers. The HTLV-1 tax mRNA load correlated with the HTLV-1 proviral DNA load ex vivo, the Tax protein expression in vitro, and the Tax-specific CD8(+) T-cell frequency ex vivo. The HTLV-1 tax mRNA load also correlated with disease severity in HAM/TSP patients. These data suggest that increased HTLV-1 expression plays an important role In the pathogenesis of HAM/TSP, and the HTLV-1 tax mRNA level could be a useful predictor of disease progression In patients with HAW TSP.

Adult neuroblastoma of the retroperitoneum and abdomen - Clinicopathologic distinction from primitive neuroectodermal tumor
T Hasegawa, T Hirose, AG Ayala, S Ito, U Tomaru, Y Matsuno, T Shimoda, S Hirohashi
AMERICAN JOURNAL OF SURGICAL PATHOLOGY, 25, 7, 918, 924, LIPPINCOTT WILLIAMS & WILKINS, Jul. 2001, [Peer-reviewed]
English, Scientific journal, Adult neuroblastoma (ANE) is a rare and poorly recognized entity among a histologically defined group of small, round-cell tumors arising in the retroperitoneum and abdomen. Eight cases of ANB were compared with seven cases of primitive neuro-ectodermal tumor (PNET) in these locations to identify clinicopathologic features that could be used to distinguish between the two lesions. The ANB study group included four men and four women 22-74 years of age (mean 38 years). Five patients with ANB presented with inflammatory symptoms or elevated levels of catecholamines and their metabolites. Five of the ANB tumors were classified as undifferentiated and three as poorly differentiated with a background of neuropil. These cases often showed immunoreactivity for multiple neural markers such as CD56, chromogranin A, synaptophysin, neurofilament, and neuron-specific enolase, but were negative for CD99, cytokeratins, desmin, myogenin, smooth muscle actin, muscle-specific actin, CD34, S-100 protein, and CD45. In contrast, all of the PNETs were positive for CD99, and four (57%) were also positive for cytokeratins. Two cases of ANB of the undifferentiated subtype had ultrastructural features characteristic of neuroblastoma and racked a chimeric transcript (EWS-FLI1or ERG), which is specific for PNET. All five patients with the undifferentiated subtype of ANB and six of the seven patients with PNET died of their disease within 3 years of discovery of the lesion. Our results show that ANB, although rare, should be considered in the differential diagnosis of patients with small, round-cell tumors in the retroperitoneum and abdomen. Appropriate immunohistochemical studies and laboratory examination enable pathologists to distinguish ANB from other differential diagnoses, especially PNET.

Combined hepatocellular carcinoma and cholangiocarcinoma growing into the common bile duct
Makoto Saito, Syuhei Hige, Hiroshi Takeda, Utano Tomaru, Masahiko Shibata, Masahiro Asaka
Journal of Gastroenterology, 36, 12, 842, 847, 2001, [Peer-reviewed]
English, Scientific journal, We report a patient with combined hepatocellular carcinoma and cholangiocarcinoma (HCC-CC) growing into the common bile duct (CBD) and showing obstructive jaundice within 2 years of the onset of the disease. The patient was a 59-year-old Japanese man in whom, at the age of 57 years, a hepatic tumor was discovered by diagnostic imaging during follow-up of hepatitis B surface antigen (HBsAg)-positive liver cirrhosis. The tumor was diagnosed as HCC. Epirubicin was injected twice, intraarterially. The patient then received oral etoposide therapy for the next 14 months. The treatment was initially effective, but approximately 2 years after the hepatic tumor was discovered, local recurrence of the tumor and a tumor thrombus in the CBD were discovered. Although he was treated with percutaneous transhepatic biliary drainage (PTBD), to reduce obstructive jaundice, the jaundice was irreversible and he died of severe hepatic failure. The autopsy findings confirmed that the hepatic tumor was HCC-CC, in which the HCC and CC components expressed α-fetoprotein (AFP) and carbohydrate antigen 19-9 (CA19-9), respectively, which accurately reflected the disease process. The underlying mechanism of the growth of HCC-CC into the CBD may differ from the underlying mechanism of the development of icteric-type HCC.

A rat model for human T lymphocyte virus type I-associated myeloneuropathy - Down-regulation of bcl-2 expression and increase in sensitivity to TNF-alpha of the spinal oligodendrocytes
XY Jiang, H Ikeda, U Tomaru, K Morita, Y Tanaka, T Yoshiki
JOURNAL OF NEUROIMMUNOLOGY, 106, 1-2, 105, 113, ELSEVIER SCIENCE BV, Jul. 2000, [Peer-reviewed]
English, Scientific journal, We reported that the tumor necrosis factor-alpha (TNF-alpha) expression and apoptotic death of oligodendrocytes appeared to be a major pathogenesis of the demyelination of spinal cords of Wistar-King-Aptekman-Hokudai (WKAH) rats with human T lymphocyte virus type I (HTLV-I) infection, HAM rats. In the present study, we examined the sensitivity to TNF-alpha-induced cell death of in vitro-separated oligodendrocytes from HTLV-I-infected WKAH rats. Although the number of non-viable oligodendrocytes increased by adding recombinant TNF-alpha, in a dose-dependent manner, in both HTLV-I-infected and uninfected control rats, oligodendrocytes from the infected rats were more susceptible to TNF-alpha. In situ detection of DNA fragmentation showed apoptotic death of oligodendrocytes. The expression of bcl-2, an anti-apoptotic gene, was strongly down-regulated in oligodendrocytes of the infected rats but not in the control rats. We suggest that the down-regulation of bcl-2 expression in the oligodendrocytes of the HTLV-I-infected rats may increase the susceptibility to TNF-alpha-induced apoptosis of oligodendrocytes, the result being development of HTLV-I-induced myeloneuropathy in rats. (C) 2000 Elsevier Science B.V. All rights reserved.

Human T-lymphocyte virus type I (HTLV-I)-induced myeloneuropathy in rats: Oligodendrocytes undergo apoptosis in the presence of HTLV-I
O Ohya, H Ikeda, U Tomaru, Yamashita, I, T Kasai, K Morita, A Wakisaka, T Yoshiki
APMIS, 108, 6, 459, 466, MUNKSGAARD INT PUBL LTD, Jun. 2000, [Peer-reviewed]
English, Scientific journal, To investigate the pathogenetic role of human T-lymphocyte virus type I (HTLV-I) in central nervous system disease, a rat model for HTLV-I-associated myelopathy/tropical spastic paraparesis, designated as HAM rat disease, was examined with regard to chronological neuropathology, from early asymptomatic phase to late disease. In the thoracic spinal cord of rats with HTLV-I infection, the first event was the appearance of apoptosis of oligodendrocytes beginning at 7 months after induced infection, thereafter followed by the appearance of white matter degeneration, increase of macrophages/activated microglia and of gemistocytic astrocytes at 12, 15 and 20 months, respectively. In the spinal cord, HTLV-I provirus DNA was evident as early as 4 months after the infection, and HTLV-I pX and the tumor necrosis factor (TNF)-alpha messages began to be expressed at age 7 months, just before or at the same time as the appearance of apoptotic cells. Collective evidence suggests that the apoptotic death of oligodendrocytes, which may be induced either directly by the local expression of HTLV-I or indirectly by TNF-alpha, through the transactive function of p40Tax, is the major cause of chronic progressive myeloneuropathy in Wistar-King-Aptekman-Hokudai rats with HTLV-I infection.

Primary lymphoma of the prostate with features of low grade B-cell lymphoma of mucosa associated lymphoid tissue: A rare cause of urinary obstruction
U Tomaru, H Ishikura, SI Kon, M Kanda, H Harada, T Yoshiki
JOURNAL OF UROLOGY, 162, 2, 496, 497, LIPPINCOTT WILLIAMS & WILKINS, Aug. 1999, [Peer-reviewed]
English, Scientific journal

A rat model of human T lymphocyte virus type I (HTLV-I) infection: In situ detection of HTLV-I provirus DNA in microglia/macrophages in affected spinal cords of rats with HTLV-1-induced chronic progressive myeloneuropathy
T Kasai, H Ikeda, U Tomaru, Yamashita, I, O Ohya, K Morita, A Wakisaka, E Matsuoka, T Moritoyo, K Hashimoto, Higuchi, I, S Izumo, M Osame, T Yoshiki
ACTA NEUROPATHOLOGICA, 97, 2, 107, 112, SPRINGER VERLAG, Feb. 1999, [Peer-reviewed]
English, Scientific journal, To investigate the pathogenetic role of human T lymphocyte virus type I (HTLV-I) in central nervous system disease, a rat model for HTLV-I-associated myelopathy/tropical spastic paraparesis, designated as HAM rat disease, has been established. Wistar-King-Aptekman-Hokudai strain rats with induced HTLV-I infection develop a chronic progressive myeloneuropathy with paraparesis of hind limbs after an incubation period of 15 months. In the affected spinal cord in these rats, white matter degeneration, demyelination and vacuolar change with microglia/macrophage infiltration are present as are the provirus DNA and the virus mRNA. To identify infected cells in the affected lesions, we carried out in situ hybridization of amplified fragments of the provirus DNA by polymerase chain reaction on thin sections, plus immunohistochemistry on the same sections. The provirus DNA was localized in some microglia/macrophages in the spinal cord lesion. In addition, the HTLV-I provirus was clearly evident not only in ED-1-negative lymphoid cells but also in ED-1-positive macrophages from lymph nodes. These observations suggest that cells of microglia/macrophage lineage may be one of dominant viral reservoirs in the spinal cords and lymph nodes in HAM rat disease. These infected microglia/macrophages may relate to cause the myeloneuropathy through neurotoxic cytokine synthesis.

Models of HTLV-I-induced diseases. Infectious transmission of HTLV-I in inbred rats and HTVL-I env-pX transgenic rats.
T Yoshiki, H Ikeda, U Tomaru, O Ohya, T Kasai, I Yamashita, K Morita, H Yamazaki, A Ishizu, Y Nakamaru, K Kikuchi, S Tanaka, A Wakisaka
Leukemia, 11 Suppl 3, 245, 6, Apr. 1997, [International Magazine]
English, Scientific journal, To examine the pathogenic roles of HTLV-I in HTLV-I-induced diseases, we developed two models; namely HTLV-I carrier rats and HTLV-I env-pX transgenic rats. Among life long HTLV-I carriers in seven rat strains, only WKAH rats with the RT1k haplotype developed chronic progressive myeloneuropathy, resembling HAM/TSP clinically and histologically in humans, designated as HAM rat disease and after long incubation periods. Apoptosis of myelin forming cells, oligodendrocytes and Schwann cells associated with HTLV-I infection appears to be the primary cause of HAM rat disease. Local activation of the pX gene and TNF alpha gene was evident in these rats. WKAH rats transgenic for HTLV-I env-pX gene were established and at age 5 weeks, swelling of the bilateral ankle joints began to develop and histological features of the affected joints resembled findings in cases of rheumatoid arthritis (RA): high-titers of rheumatoid factors were present in these rats. A series of vascular collagen diseases such as polyarteritis nodosa-like angiitis, polymyositis, myocarditis, and Sjögren's syndrome-like sialodenitis together with RA were present, even in one individual animal. These transgenic rats as well as HAM rats appear to be suitable animal models for elucidating pathogenic mechanisms implicated in HTLV-I-induced diseases and also various demyelinating vascular collagen diseases of unknown etiology.

皮膚血管炎動物モデルの完成
川上民裕, 中出一生, 田村宥人, 伊藤吹夕, 西端友香, 益田紗季子, 外丸詩野, 石津明洋, 脈管学(Web), 63, 1, 2023

正常ラットに抗PSPT抗体と抗LAMP2抗体の静脈注射により皮膚血管炎の発症に成功した
川上民裕, 中出一生, 田村宥人, 西端友香, 益田紗季子, 石津明洋, 伊藤吹夕, 外丸詩野, 日本皮膚科学会雑誌, 133, 3, 2023

人工知能による結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の鑑別
新海隼人, 加藤千恵次, 川上民裕, 高橋啓, 西端友香, 益田紗季子, 田中敏, 外丸詩野, 石津明洋, 脈管学(Web), 61, 1, 2021

myosin light chain6を認議する抗好中球細胞外トラップ(NETs)抗体はNETs分解阻害活性を持つ
西端友香, 益田紗季子, 外丸詩野, 石津明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 65th, 2021

Effects of hypoxia on neutrophil extracellular trap formation
益田紗季子, 西端友香, 田中敏, 外丸詩野, 辻野一三, 石津明洋, 日本病理学会会誌, 110, 1, 2021

Recombinant thrombomodulin ameliorates autoimmune vasculitis via immune response regulation and tissue injury protection.
Kanako Watanabe-Kusunoki, Daigo Nakazawa, Yoshihiro Kusunoki, Takashi Kudo, Fumihiko Hattanda, Saori Nishio, Sakiko Masuda, Utano Tomaru, Takeshi Kondo, Tatsuya Atsumi, Akihiro Ishizu, Journal of autoimmunity, 108, 102390, 102390, Mar. 2020, [International Magazine]
Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is characterized by necrotizing vasculitis with the presence of pathogenic ANCA. ANCA can potentially cause neutrophil activation and induce neutrophil extracellular traps (NETs), resulting in endothelial damage as well as activation of autoreactive B cells and alternative complement pathway. Recombinant thrombomodulin (rTM) protects the endothelium from vascular injury during disseminated intravascular coagulation, thus we hypothesized that rTM ameliorates necrotizing vasculitis in AAV. In this study, rTM was administered in an experimental AAV rat model. Treatment of experimental AAV rats with rTM improved pulmonary hemorrhage and glomerulonephritis, with a suppression of ANCA production and NETs formation. In addition, in vitro experiments showed that rTM bound to neutrophils via Mac-1 (macrophage-1 antigen) and inhibited ANCA-induced NETs formation accompanied by a suppression of histone citrullination, leading to a protection of the endothelium from NETs toxicity. Additionally, rTM affected lymphocytes leading to the inhibition of pro-inflammatory cytokine/chemokin in PBMC during the antibody production process, which might indirectly be involved in the reduction of pathogenic ANCA. Our data revealed that the rTM could ameliorate autoimmune vasculitis through a combination of different biological mechanisms., English

抗GBM抗体が認識するエピトープの表出に関する検討
西端友香, 野々川茉佑, 益田紗季子, 中沢大悟, 田中敏, 外丸詩野, 石津明洋, 脈管学(Web), 60, 2, 2020

HIGH DOSE IMMUNOGLOBULINS CAN INHIBIT NEUTROPHIL EXTRACELLULAR TRAP FORMATION, EVENTUALLY PREVENT THE DEVELOPMENT OF MPO-ANCA-ASSOCIATED VASCULITIS
Ryo Uozumi, Sakiko Masuda, Yuka Nishibata, Shun Tanimura, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu, RHEUMATOLOGY, 58, Mar. 2019
OXFORD UNIV PRESS, English, Summary international conference

抗糸球体基底膜抗体病(抗GBM病)の頻回再発症例の抗GBMが認識するエピトープ
西端友香, 東里緒, 益田紗季子, 中沢大悟, 田中敏, 外丸詩野, 中林公正, 石津明洋, 脈管学(Web), 59, 3, 2019

シクロフィリンDをターゲットとしたANCA関連壊死性血管炎に対する新規治療薬の開発
工藤孝司, 中沢大悟, 白鳥里佳, 楠加奈子, 西尾妙織, 益田紗季子, 外丸詩野, 石津明洋, 渥美達也, 日本臨床免疫学会総会プログラム・抄録集, 47th, 2019

ヒストンは好中球細胞上にLAMP2を表出し,抗LAMP2抗体と連携して皮膚小血管炎の発症機序に関与している
川上民裕, 竹内そら, 菊池彩翔, 西端友香, 益田紗季子, 外丸詩野, 石津明洋, 脈管学(Web), 59, 3, 2019

EFFECTS OF RECOMBINANT THROMBOMODULIN ON EXPERIMENTAL AUTOIMMUNE VASCULITIS VIA THE INHIBITION OF NEUTROPHIL EXTRACELLULAR TRAPS
Kanako Watanabe, Daigo Nakazawa, Yoshihiro Kusunoki, Fumihiko Hattanda, Saori Nishio, Sakiko Masuda, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu, NEPHROLOGY DIALYSIS TRANSPLANTATION, 33, 34, 34, May 2018
OXFORD UNIV PRESS, English, Summary international conference

ANCA関連血管炎(AAV)の壊死性病変部における好中球細胞外トラップ(NETs)の存在と病的意義　　　　　　　　　　　　　　　
益田 紗季子, 野々川 茉佑, 西端 友香, 岩崎 沙理, 辻 隆裕, 田中 敏, 外丸 詩野, 川上 民裕, 石津 明洋, 日本病理学会会誌, 107, 1, 330, 330, Apr. 2018
(一社)日本病理学会, Japanese

免疫グロブリン大量静注療法(IVIG)は好中球細胞外トラップ(NETs)の抑制によりMPO-ANCA関連血管炎を抑制する　　　　　　　　　　　　　　　
魚住 諒, 井口 理彩, 益田 紗季子, 西端 友香, 谷村 瞬, 中沢 大悟, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 107, 1, 331, 331, Apr. 2018
(一社)日本病理学会, Japanese

致死的な腹腔内出血を来した顕微鏡的多発血管炎の一剖検例　　　　　　　　　　　　　　　
伊丹 久実, 木内 隆之, 外丸 詩野, 大塚 紀幸, 田中 敏, 石津 明洋, 笠原 正典, 日本病理学会会誌, 107, 1, 535, 535, Apr. 2018
(一社)日本病理学会, Japanese

血管炎1:血管炎のバイオマーカー・病態解析 免疫グロブリン製剤は好中球細胞外トラップ(NETs)の形成抑制を介してMPO-ANCA関連血管炎(MPO-AAV)の発症を抑制する　　　　　　　　　　　　　　　
魚住 諒, 益田 紗季子, 西端 友香, 谷村 瞬, 中沢 大悟, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 62回, 433, 433, Mar. 2018
(一社)日本リウマチ学会, Japanese

リウマチ性疾患の画像2:関節エコーその他 超高周波プローブを用いたSuperb Micro-vascular Imagingによる超音波検査でのラット足関節の早期関節炎診断　　　　　　　　　　　　　　　
西田 睦, 谷村 瞬, 神島 保, 西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 渥美 達也, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 62回, 485, 485, Mar. 2018
(一社)日本リウマチ学会, Japanese

スタチンの関節炎抑制効果とその機序の解明　　　　　　　　　　　　　　　
谷村 瞬, 西田 睦, 神島 保, 西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 渥美 達也, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 62回, 778, 778, Mar. 2018
(一社)日本リウマチ学会, Japanese

BDNFはHUVECにおけるangiogeninの分泌と核移行を誘導する
森綾子, 西岡祐介, 山田真衣, 西端友香, 益田紗季子, 外丸詩野, 本間直幸, 森山隆則, 石津明洋, 日本血管生物医学会学術集会プログラム・抄録集, 26th, 2018

スタチンの関節炎抑制効果と機序の検討
谷村瞬, 渥美達也, 西田睦, 堀江達則, 神島保, 玉井絵里香, 森村豊, 西端友香, 益田紗季子, 石津明洋, 外丸詩野, 北海道医学雑誌, 93, 2, 2018

プロテアーゼによる抗糸球体基底膜抗体病の隔絶抗原の表出
西端友香, 植松千浩, 益田紗季子, 田中敏, 外丸詩野, 石津明洋, 脈管学(Web), 58, 3, 2018

ANCA関連血管炎の壊死性病変部におけるNETsの存在と病的意義
益田紗季子, 野々川茉佑, 西端友香, 岩崎沙理, 辻隆裕, 田中敏, 外丸詩野, 川上民裕, 石津明洋, 脈管学(Web), 58, 3, 2018

原発性肺扁平上皮癌におけるPD-L1とNT5E(CD73)の発現と予後に関する検討　　　　　　　　　　　　　　　
桑原 健, 畑中 豊, 畑中 佳奈子, 清水 知浩, 岡田 宏美, 加賀 基知三, 樋田 泰浩, 秋田 弘俊, 榊原 純, 外丸 詩野, 松野 吉宏, 日本癌学会総会記事, 76回, P, 3259, Sep. 2017
日本癌学会, English

原発性肺扁平上皮癌におけるPD-L1とNT5E(CD73)の発現と予後に関する検討　　　　　　　　　　　　　　　
桑原 健, 畑中 豊, 畑中 佳奈子, 清水 知浩, 岡田 宏美, 加賀 基知三, 樋田 泰浩, 秋田 弘俊, 榊原 純, 外丸 詩野, 松野 吉宏, 日本癌学会総会記事, 76回, P, 3259, Sep. 2017
日本癌学会, English

病理診断に直結した組織学 VI.胸郭,漿膜 2.胸腺
中智昭, 外丸詩野, 松野吉宏, 病理と臨床, 35, 151‐160, 11 Apr. 2017
Japanese

プロテアソーム機能低下モデルマウスにおける脳機能障害の解析　　　　　　　　　　　　　　　
外丸 詩野, 伊藤 智樹, 大村 優, 石津 明洋, 笠原 正典, 日本病理学会会誌, 106, 1, 323, 323, Mar. 2017
(一社)日本病理学会, Japanese

フローサイトメトリーによる定量的好中球細胞外トラップ測定法　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 松尾 淳司, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 106, 1, 350, 350, Mar. 2017
(一社)日本病理学会, Japanese

_7 RAT TYPE II NKT CELL CLONE PATHOGENIC FOR SMALL VESSEL VASCULITIS RECOGNIZES STEROL CARRIER PROTEIN 2
Yusuke Nishioka, Madoka Yamaguchi, Ai Kawakami, Maya Munehiro, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu, RHEUMATOLOGY, 56, 30, 31, Mar. 2017
OXFORD UNIV PRESS, English, Summary international conference

THE EFFECT OF PEPTIDYLARGININE DEIMINASE INHIBITOR ON NET FORMATION AND MPO-ANCA PRODUCTION IN MOUSE MODEL
Yoshihiro Kusunoki, Daigo Nakazawa, Haruki Shida, Fumihiko Hattanda, Arina Miyoshi, Sakiko Masuda, Saori Nishio, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu, RHEUMATOLOGY, 56, 112, 113, Mar. 2017
OXFORD UNIV PRESS, English, Summary international conference

自己免疫疾患とNETosis
楠由宏, 益田紗季子, 外丸詩野, 石津明洋, 月刊リウマチ科, 57, 4, 2017

ANCA関連血管炎(AAV)における抗NETs抗体の存在と病的意義
八反田文彦, 楠由宏, 志田玄貴, 中沢大悟, 西尾妙織, 益田紗季子, 外丸詩野, 渥美達也, 石津明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 61st, 2017

スタチン製剤の関節炎抑制効果と機序の検討
谷村瞬, 渥美達也, 西田睦, 堀江達則, 神島保, 齋藤克己, 西端友香, 益田紗季子, 石津明洋, 外丸詩野, 北海道医学雑誌, 92, 2, 2017

抗PSPT抗体による抗リン脂質抗体症候群動物モデルの完成
川上民裕, 石津明洋, 益田紗季子, 外丸詩野, 日本リウマチ学会総会・学術集会プログラム・抄録集, 61st, 2017

抗PSPT抗体は,正常ラットに血栓を発症させる
川上民裕, 山田真衣, 高島滉平, 西岡佑介, 西端友香, 益田紗季子, 吉田繁, 外丸詩野, 石津明洋, 脈管学(Web), 57, 2, 2017

プロテアソームとCD8+ T細胞レパトアの形成　　　　　　　　　　　　　　　
外丸 詩野, 石津 明洋, 宮島 祥太, 木内 静香, 笠原 正典, MHC: Major Histocompatibility Complex, 23, 2Suppl., 95, 95, Oct. 2016
日本組織適合性学会, Japanese

CD8+ T細胞の胸腺選択にプロテアソームβ5サブユニットが与える影響　　　　　　　　　　　　　　　
宮島 祥太, 外丸 詩野, 石津 明洋, 木内 静香, 笠原 正典, 日本病理学会会誌, 105, 1, 375, 375, Apr. 2016
(一社)日本病理学会, Japanese

胸腺上皮性腫瘍におけるカテプシンV及びカテプシンSの発現　　　　　　　　　　　　　　　
木内 静香, 外丸 詩野, 石津 明洋, 大塚 紀幸, 今川 誠, 岩崎 沙理, 鈴木 昭, 丸川 活司, 松野 吉宏, 笠原 正典, 日本病理学会会誌, 105, 1, 471, 471, Apr. 2016
(一社)日本病理学会, Japanese

非小細胞肺癌における免疫プロテアソームサブユニットβ5iの発現　　　　　　　　　　　　　　　
木内 隆之, 外丸 詩野, 石津 明洋, 今川 誠, 岩崎 沙理, 鈴木 昭, 松野 吉宏, 笠原 正典, 日本病理学会会誌, 105, 1, 534, 534, Apr. 2016
(一社)日本病理学会, Japanese

肺移植施行後に日和見感染症により死亡に至った肺ランゲルハンス細胞組織球症の一例　　　　　　　　　　　　　　　
四宮 万里絵, 松林 里佳, 外丸 詩野, 木内 隆之, 大塚 紀幸, 石津 明洋, 笠原 正典, 日本病理学会会誌, 105, 1, 585, 585, Apr. 2016
(一社)日本病理学会, Japanese

Peptigylarginine deiminase 4阻害薬は好中球細胞外トラップの形成阻害を介してMPO-ANCA産生を抑制する
楠由宏, 中沢大悟, 志田玄貴, 八反田文彦, 益田紗季子, 外丸詩野, 西尾妙織, 渥美達也, 石津明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 60th, 2016

MPO-ANCA関連血管炎におけるNETs-ANCA悪循環
志田玄貴, 八反田文彦, 三次有奈, 楠由宏, 中沢大悟, 佐藤遥, 橋本展洋, 林晃正, 益田紗季子, 石津明洋, 外丸詩野, 北海道医学雑誌, 91, 2, 2016

抗ラクトフェリン抗体は好酸球性多発血管炎性肉芽腫症において好中球細胞外トラップの形成を促進し,疾患活動性に関与する
志田玄貴, 中沢大悟, 八反田文彦, 楠由宏, 益田紗季子, 外丸詩野, 川上民裕, 渥美達也, 石津明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 60th, 2016

自己血管内皮細胞反応性小型血管炎惹起性type II NKT細胞が認識する分子の同定
西岡佑介, 山口まどか, 川上愛, 宗廣真矢, 山田真衣, 益田紗季子, 外丸詩野, 石津明洋, 日本病理学会会誌, 105, 1, 2016

Proteasomes and age-related disorders
外丸 詩野, 笠原 正典, 細胞, 47, 12, 586, 589, Nov. 2015
ニューサイエンス社, Japanese

The Impact of c-Fos/Activator Protein-1 Inhibition on Allogeneic Pancreatic Islet Transplantation
T. Yoshida, K. Yamashita, M. Watanabe, Y. Koshizuka, D. Kuraya, M. Ogura, Y. Asahi, H. Ono, S. Emoto, T. Mizukami, N. Kobayashi, S. Shibasaki, U. Tomaru, H. Kamachi, M. Matsushita, S. Shiozawa, S. Hirono, S. Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 15, 10, 2565, 2575, Oct. 2015
Unpreventable allograft rejection is one of the main problems in pancreatic islet transplantation (PIT). Therefore, it is imperative to develop a more effective immunosuppressive strategy. The blockade of transcription factors has been a central part of T celldepleting immunosuppressive therapies, as typified by the use of calcineurin inhibitors. The inhibition of activator protein-1 (AP-1) offers a novel strategy for immunosuppression in PIT, although to date, no reports on the effects of AP-1 inhibition are available. In this study, we investigated the immunosuppressive effects of T-5224, a c-Fos/AP-1-selective inhibitor, on murine T cells activated by alpha CD3+alpha CD28 mAbs. T-5224 inhibited proliferation, CD25 up-regulation, and the production of IL-2 and interferon-gamma. In addition, T-5224 blocked the nuclear translocation of c-Fos/AP-1 in activated murine T cells. In BALB/c (H-2(d))-to-C57BL/6J (H-2(b)) mouse PIT, the 2-week administration of T-5224 prolonged survival of 600 islet allografts in a dose-dependent manner. When combined with a 2-week low-dose tacrolimus, the T-5224 treatment markedly prolonged allograft survival to over 300 days, while the efficacy was indeterminate when transplanted islet allograft mass was reduced to 300. We conclude that the c-Fos/AP-1 inhibition by T-5224 is a potentially attractive strategy for allogeneic PIT., WILEY-BLACKWELL, English

Proteasome Inhibition by Bortezomib Prevents Early Graft Failure After Pancreatic Islet Transplantation
H. Ono, Y. Asahi, T. Yoshida, Y. Koshizuka, M. Watanabe, U. Tomaru, S. Emoto, M. Fukai, A. Taketomi, S. Todo, K. Yamashita, AMERICAN JOURNAL OF TRANSPLANTATION, 15, May 2015
WILEY-BLACKWELL, English, Summary international conference

膵島移植におけるプロテアソーム阻害による早期グラフト障害の抑制
小野仁, 旭火華, 吉田雅, 腰塚靖之, 渡辺正明, 外丸詩野, 江本慎, 深井原, 嶋村剛, 武冨紹信, 藤堂省, 山下健一郎, 日本外科学会定期学術集会(Web), 115回, OP, 7, Apr. 2015
(一社)日本外科学会, Japanese

MPO-ANCA関連血管炎に対するPAD4阻害薬の効果　　　　　　　　　　　　　　　
楠 由宏, 中沢 大悟, 三次 有奈, 志田 玄貴, 外丸 詩野, 西尾 妙織, 渥美 達也, 石津 明洋, 日本腎臓学会誌, 57, 3, 507, 507, Apr. 2015
(一社)日本腎臓学会, Japanese

高血糖による好中球細胞外トラップ(neutrophil extracellular traps:NETs)の形成亢進　　　　　　　　　　　　　　　
三次 有奈, 山田 真衣, 舘山 ゆう, 楠 由宏, 志田 玄貴, 中沢 大悟, 中村 昭伸, 外丸 詩野, 三好 秀明, 渥美 達也, 石津 明洋, 糖尿病, 58, Suppl.1, S, 392, Apr. 2015
(一社)日本糖尿病学会, Japanese

その他のアレルギー疾患・免疫疾患 リウマトイド血管炎患者とモデルラットで認めた血中抗ホスファチジルセリン・プロトロンビン複合体抗体上昇　　　　　　　　　　　　　　　
川上 民裕, 竹内 そら, 相馬 良直, 川上 愛, 外丸 詩野, 石津 明洋, アレルギー, 64, 3-4, 490, 490, Apr. 2015
(一社)日本アレルギー学会, Japanese

悪性腹膜中皮腫との鑑別に苦慮したatypical mesothelial proliferationの1例　　　　　　　　　　　　　　　
石川 麻美, 堀井 恒哉, 小山内 翔祐, 勘野 真紀, 大塚 紀幸, 外丸 詩野, 石津 明洋, 高木 芳武, 日本臨床細胞学会雑誌, 54, Suppl.1, 279, 279, Apr. 2015
(公社)日本臨床細胞学会, Japanese

ANCA関連血管炎と好中球細胞外トラップ(neutrophil extracellular traps:NETs)を正しく理解する 抗好中球細胞質抗体(ANCA) 好中球細胞外トラップ(NETs)の悪循環　　　　　　　　　　　　　　　
中沢 大悟, 楠 由宏, 志田 玄貴, 西尾 妙織, 外丸 詩野, 渥美 達也, 石津 明洋, 脈管学, 55, 2, 35, 35, Mar. 2015
(一社)日本脈管学会, Japanese

リウマトイド血管炎を発症した患者とモデルラットでは血中抗ホスファチジルセリン・プロトロンビン複合体抗体が上昇している　　　　　　　　　　　　　　　
川上 民裕, 竹内 そら, 相馬 良直, 川上 愛, 外丸 詩野, 石津 明洋, 脈管学, 55, 2, 40, 40, Mar. 2015
(一社)日本脈管学会, Japanese

血管炎 動物モデル全身に血栓形成を誘導させる新規抗リン脂質抗体の作成に成功した　　　　　　　　　　　　　　　
川上 民裕, 志田 玄貴, 中沢 大悟, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 59回, 351, 351, Mar. 2015
(一社)日本リウマチ学会, Japanese

血管炎 抗ラクトフェリン抗体の病原性　　　　　　　　　　　　　　　
志田 玄貴, 中沢 大悟, 外丸 詩野, 川上 民裕, 渥美 達也, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 59回, 351, 351, Mar. 2015
(一社)日本リウマチ学会, Japanese

胸腺T細胞選択におけるプロテアソームキモトリプシン様活性サブユニットの役割　　　　　　　　　　　　　　　
宮島 祥太, 外丸 詩野, 石津 明洋, 木内 静香, 大井 智貴, 笠原 正典, 日本病理学会会誌, 104, 1, 284, 284, Mar. 2015
(一社)日本病理学会, Japanese

プロテアソーム機能低下マウスにおける記銘力障害の解析　　　　　　　　　　　　　　　
伊藤 智樹, 外丸 詩野, 大村 優, 戸松 留花, 石津 明洋, 笠原 正典, 日本病理学会会誌, 104, 1, 322, 322, Mar. 2015
(一社)日本病理学会, Japanese

高血糖による好中球細胞外トラップ(neutrophil extracellular traps:NETs)の形成亢進　　　　　　　　　　　　　　　
三次 有奈, 山田 真衣, 舘山 ゆう, 楠 由宏, 志田 玄貴, 中沢 大悟, 外丸 詩野, 三好 秀明, 渥美 達也, 石津 明洋, 日本病理学会会誌, 104, 1, 379, 379, Mar. 2015
(一社)日本病理学会, Japanese

ダウン症患者の胸腺におけるプロテアソームサブユニットβ5tの発現低下　　　　　　　　　　　　　　　
木内 静香, 外丸 詩野, 辻 隆裕, 石津 明洋, 鈴木 昭, 大塚 紀幸, 伊藤 智樹, 池田 仁, 深澤 雄一郎, 笠原 正典, 日本病理学会会誌, 104, 1, 460, 460, Mar. 2015
(一社)日本病理学会, Japanese

壁在結節を伴う卵巣粘液産生腺癌の3例　　　　　　　　　　　　　　　
河内 麻里亜, 横山 達也, 大塚 紀幸, 藤本 俊郎, 池田 仁, 高木 芳武, 石津 明洋, 外丸 詩野, 笠原 正典, 日本病理学会会誌, 104, 1, 522, 522, Mar. 2015
(一社)日本病理学会, Japanese

TypeB胸腺腫、胸腺癌におけるカテプシンの発現　　　　　　　　　　　　　　　
安部 樹太朗, 有賀 茜, 外丸 詩野, 木内 静香, 石津 明洋, 大塚 紀幸, 清水 知浩, 丸川 活司, 松野 吉宏, 笠原 正典, 日本病理学会会誌, 104, 1, 528, 528, Mar. 2015
(一社)日本病理学会, Japanese

Churg-Strauss症候群による多発小腸穿孔により死亡した症例　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 江辺 広志, 浄土 智, 藤咲 淳, 脈管学, 54, 12, 208, 208, Dec. 2014
(一社)日本脈管学会, Japanese

MPO-ANCA関連血管炎患者におけるneutrophil extracelular traps(NETs)の制御異常　　　　　　　　　　　　　　　
中沢 大悟, 志田 玄貴, 西尾 妙織, 渥美 達也, 吉田 雅治, 外丸 詩野, 石津 明洋, 脈管学, 54, 12, 221, 221, Dec. 2014
(一社)日本脈管学会, Japanese

鑑別に苦慮した胸腺腫瘍の1例　　　　　　　　　　　　　　　
桑原 健, 岩崎 沙理, 桑原 博昭, 外丸 詩野, 笠原 正典, 石津 明洋, 松野 吉宏, 鈴木 昭, 日本病理学会会誌, 103, 2, 46, 46, Sep. 2014
(一社)日本病理学会, Japanese

DECREASED FREQUENCY OF PERIPHERAL AND INTESTINAL NKG2A-POSITIVE T CELLS IN ULCERATIVE COLITIS PATIENTS
W. Kobayashi, T. Katsurada, U. Tomaru, N. Sakamoto, M. Kasahara, GUT, 63, A177, A177, Jun. 2014
BMJ PUBLISHING GROUP, English, Summary international conference

顕微鏡的多発血管炎と血栓症はMPO-ANCAと好中球細胞外トラップを介して関連する　　　　　　　　　　　　　　　
今本 鉄平, 中沢 大悟, 志田 玄貴, 鈴木 昭, 大塚 紀幸, 外丸 詩野, 石津 明洋, 北海道医学雑誌, 89, 1, 91, 91, May 2014
北海道医学会, Japanese

ENHANCED FORMATION AND DISORDERED REGULATION OF NETS IN MPO-ANCA-ASSOCIATED VASCULITIS
Nakazawa Daigo, Shida Haruki, Tomaru Utano, Yoshida Masaharu, Nishio Saori, Atsumi Tatsuya, Ishizu Akihiro, NEPHROLOGY, 19, 149, 150, May 2014
WILEY-BLACKWELL, English, Summary international conference

胸腺におけるプロテアソームキモトリプシン様活性サブユニットの発現とT細胞選択　　　　　　　　　　　　　　　
木内 静香, 外丸 詩野, 紺野 沙織, 石津 明洋, 宮島 祥太, 平川 彩香, 笠原 正典, 日本病理学会会誌, 103, 1, 247, 247, Mar. 2014
(一社)日本病理学会, Japanese

高齢男性でみられたEBV陽性肝脾γδT細胞リンパ腫の一例　　　　　　　　　　　　　　　
竹中 淳規, 大塚 紀幸, 藤田 裕美, 中馬 誠, 外丸 詩野, 石津 明洋, 笠原 正典, 日本病理学会会誌, 103, 1, 407, 407, Mar. 2014
(一社)日本病理学会, Japanese

血管炎症候群の新しい考え方(第7回) 血管炎の新たな発症機構 「好中球の網」NETs　　　　　　　　　　　　　　　
中沢 大悟, 志田 玄貴, 西尾 妙織, 渥美 達也, 吉田 雅治, 外丸 詩野, 石津 明洋, 分子リウマチ治療, 7, 1, 34, 38, Jan. 2014
好中球の殺菌機構であるneutrophil extracellular traps(NETs)がさまざまな自己免疫疾患と関連することが指摘されるようになったが、そのなかでもとくに抗好中球細胞質抗体(ANCA)産生を特徴とする顕微鏡的多発血管炎(MPA)が密接な関係にある。NETsは、殺菌蛋白やDNAを細胞外に放出して微生物を捕捉・殺菌する強力な免疫機構であるが、ANCAは、感染微生物がなくとも好中球に作用してNETsを誘導する。このNETs誘導活性は、血管炎の活動性に比例し、また誘導されたNETsは、本疾患患者ではさまざまな原因で制御されない背景にある。すなわち、ANCAはNETsを誘導して血管炎を引き起こし、そのNETsはANCAの対応抗原を含んだまま制御不能で存続するというNETsとANCAを介した悪循環が、MPAの病態を形成している。(著者抄録), (株)先端医学社, Japanese

von Recklinghausen病による動脈破裂の病理組織学的検討　　　　　　　　　　　　　　　
木内 隆之, 外丸 詩野, 高田 明生, 石津 明洋, 脈管学, 53, December, 222, 223, Dec. 2013
(一社)日本脈管学会, Japanese

MPO-ANCAのaffinityとNETs誘導率はMPO-ANCA関連血管炎の疾患活動性を反映する　　　　　　　　　　　　　　　
中沢 大悟, 石川 康暢, 柴崎 跡也, 西尾 妙織, 渥美 達也, 外丸 詩野, 吉田 雅治, 石津 明洋, 北海道医学雑誌, 88, 4-5, 160, 160, Sep. 2013
北海道医学会, Japanese

血管炎症候群の新しい考え方(第4回)血管炎の予後は予測できるか : 予測因子検索の道
石津 明洋, 外丸 詩野, 村井 太一, 分子リウマチ治療, 6, 2, 82, 86, May 2013
先端医学社, Japanese

炎症・免疫機構の新基軸と疾病の病理 好中球細胞外トラップ(NETs)の異常とMPO-ANCA関連血管炎の発症　　　　　　　　　　　　　　　
中沢 大悟, 外丸 詩野, 西尾 妙織, 渥美 達也, 笠原 正典, 石津 明洋, 日本病理学会会誌, 102, 1, 185, 185, Apr. 2013
(一社)日本病理学会, Japanese

血管炎の発症メカニズム 自己血管内皮細胞反応性NKT細胞による血管炎発症モデル　　　　　　　　　　　　　　　
川上 愛, 飯沼 千景, 脇 雅, 山口 まどか, 外丸 詩野, 笠原 正典, 吉木 敬, 石津 明洋, 日本病理学会会誌, 102, 1, 198, 198, Apr. 2013
(一社)日本病理学会, Japanese

プロテアソームの発現異常における免疫応答の変化　　　　　　　　　　　　　　　
紺野 沙織, 外丸 詩野, 岸本 栞奈, 石津 明洋, 笠原 正典, 日本病理学会会誌, 102, 1, 330, 330, Apr. 2013
(一社)日本病理学会, Japanese

顕微鏡的多発血管炎と血栓症はMPO-ANCAと好中球細胞外トラップを介して関連する　　　　　　　　　　　　　　　
今本 鉄平, 中沢 大悟, 大塚 紀幸, 外丸 詩野, 笠原 正典, 石津 明洋, 日本病理学会会誌, 102, 1, 502, 502, Apr. 2013
(一社)日本病理学会, Japanese

HELLP症候群に合併したhepatic ruptureの一剖検例　　　　　　　　　　　　　　　
山本 岳, 藤井 真理子, 大塚 紀幸, 光部 兼六郎, 櫻井 宏治, 外丸 詩野, 石津 明洋, 笠原 正典, 日本病理学会会誌, 102, 1, 502, 502, Apr. 2013
(一社)日本病理学会, Japanese

リツキシマブ投与後に日和見感染症を併発した顕微鏡的多発血管炎の一剖検例　　　　　　　　　　　　　　　
味藤 静, 外丸 詩野, 大塚 紀幸, 石津 明洋, 笠原 正典, 日本病理学会会誌, 102, 1, 503, 503, Apr. 2013
(一社)日本病理学会, Japanese

潰瘍性大腸炎における末梢血および大腸粘膜固有層NKG2A+ T細胞の減少
桂田 武彦, 小林 和夏, 外丸 詩野, 馬場 智久, 古川 滋, 石津 明洋, 竹田 和由, 坂本 直哉, 浅香 正博, 武田 宏司, 笠原 正典, 北海道醫學雜誌 = Acta medica Hokkaidonensia, 88, 2, 104, 104, 01 Apr. 2013
Japanese

Abundant neutrophil extracellular traps in thrombus of patient with microscopic polyangiitis
D. Nakazawa, U. Tomaru, S. Jodo, S. Nishio, T. Atsumi, A. Ishizu, PRESSE MEDICALE, 42, 4, 755, 755, Apr. 2013
MASSON EDITEUR, English, Summary international conference

Prediction of outcome of treatment by gene expression profiling of peripheral blood in patients with microscopic polyangiitis
A. Ishizu, U. Tomaru, T. Murai, T. Yamamoto, T. Atsumi, T. Yoshiki, H. Makino, S. Ozaki, PRESSE MEDICALE, 42, 4, 733, 733, Apr. 2013
MASSON EDITEUR, English, Summary international conference

血管炎 顕微鏡的多発血管炎(MPA)患者に合併した深部静脈血栓における過剰なNETs形成　　　　　　　　　　　　　　　
中沢 大悟, 外丸 詩野, 浄土 智, 渥美 達也, 石津 明洋, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 57回・22回, 434, 434, Mar. 2013
(一社)日本リウマチ学会, Japanese

プロテアソーム機能不全による網膜変性　　　　　　　　　　　　　　　
安藤 亮, 野田 航介, 外丸 詩野, 野田 実香, 高階 沙織, 董 陽子, 木下 哲志, 福原 淳一, 董 震宇, 神田 敦宏, 石田 晋, 日本眼科学会雑誌, 117, 臨増, 312, 312, Mar. 2013
(公財)日本眼科学会, Japanese

【血管炎-基礎と臨床のクロストーク-】 ANCA関連血管炎の病因・病理、診断・治療 ANCA関連血管炎(AAV)の基礎研究から臨床へのアプローチ ANCA関連血管炎の病因とそのバイオマーカー プロピルチオウラシルとNETsの異常形成・分解異常　　　　　　　　　　　　　　　
中沢 大悟, 外丸 詩野, 西尾 妙織, 渥美 達也, 石津 明洋, 日本臨床, 71, 増刊1 血管炎, 244, 249, Feb. 2013
(株)日本臨床社, Japanese

【血管炎-基礎と臨床のクロストーク-】 最新の研究トピックス AP-VAS 2012から 血管炎の基礎研究 自己反応性NKT細胞と血管炎　　　　　　　　　　　　　　　
飯沼 千景, 脇 雅, 山口 まどか, 外丸 詩野, 石津 明洋, 日本臨床, 71, 増刊1 血管炎, 502, 505, Feb. 2013
(株)日本臨床社, Japanese

深部静脈血栓症を伴った顕微鏡的多発血管炎(MPA)の一剖検例 MPAと血栓の関連について　　　　　　　　　　　　　　　
中沢 大悟, 外丸 詩野, 山本 知穂, 浄土 智, 石津 明洋, 北海道医学雑誌, 87, 4-5, 212, 212, Aug. 2012
北海道医学会, Japanese

プロテアソームの機能低下は老化, 脂肪代謝の異常を引き起こす
外丸 詩野, 高橋 里美, 石津 明洋, 宮武 由甲子, 合田 文, 鈴木 小百合, 小野 綾子, 小原 次郎, 馬場 智久, 村田 茂穂, 田中 啓二, 笠原 正典, 北海道醫學雜誌 = Acta medica Hokkaidonensia, 87, 4, 207, 207, 01 Aug. 2012
Japanese

Presence of anti-LAMP-2 antibody induces cutaneous vasculitis of rats
Tamihiro Kawakami, Sora Takeuchi, Satoko Kimura, Yoshinao Soma, Masashi Waki, Madoka Yamaguchi, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu, JOURNAL OF DERMATOLOGY, 39, 49, 49, Jun. 2012
WILEY-BLACKWELL, English, Summary international conference

ABNORMAL CONFORMATION AND IMPAIRED DEGRADATION OF NEUTROPHIL EXTRACELLULAR TRAPS INDUCED BY PROPYLTHIOURACIL ARE IMPLICATED IN THE PATHOGENESIS OF MPO-ANCA-ASSOCIATED VASCULITIS
Daigo Nakazawa, Saori Nishio, Sekiya Shibasaki, Utano Tomaru, Ishizu Akihiro, NEPHROLOGY DIALYSIS TRANSPLANTATION, 27, 442, 442, May 2012
OXFORD UNIV PRESS, English, Summary international conference

Plasma-dependent, antibody- and Fcγ receptor-mediated translocation of CD8 molecules from T cells to monocytes
岩崎 沙理, 益田 紗季子, 馬場 智久, 外丸 詩野, 勝俣 一晃, 笠原 正典, 石津 明洋, 北海道醫學雜誌 = Acta medica Hokkaidonensia, 87, 2, 68, 68, 01 Apr. 2012
Japanese

血管炎(2) 自己血管内皮細胞反応性NKT細胞による血管炎発症モデル　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56回・21回, 330, 330, Mar. 2012
(一社)日本リウマチ学会, Japanese

血管炎(2) プロピルチオウラシルによるNETs(neutrophil extracellular traps)の形成および分解障害とMPO-ANCAの産生 MPO-ANCA関連血管炎の発症機序　　　　　　　　　　　　　　　
中沢 大悟, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56回・21回, 331, 331, Mar. 2012
(一社)日本リウマチ学会, Japanese

プロテアソーム活性の低下が腫瘍微小環境に与える影響 腫瘍増殖の抑制効果について　　　　　　　　　　　　　　　
小野 綾子, 外丸 詩野, 石津 明洋, 小原 次郎, 紺野 沙織, 笠原 正典, 日本病理学会会誌, 101, 1, 283, 283, Mar. 2012
(一社)日本病理学会, Japanese

成人T細胞白血病(ATL)の病態における上皮細胞の役割の検討　　　　　　　　　　　　　　　
宮武 由甲子, 外丸 詩野, Sheehy Noreen, 石津 明洋, Hall William W, 笠原 正典, 日本病理学会会誌, 101, 1, 291, 291, Mar. 2012
(一社)日本病理学会, Japanese

TNFα変換酵素(TACE)の過剰発現が糖および脂質代謝へ及ぼす影響　　　　　　　　　　　　　　　
松井 由希, 深谷 進司, 外丸 詩野, 渥美 達也, 笠原 正典, 石津 明洋, 日本病理学会会誌, 101, 1, 314, 314, Mar. 2012
(一社)日本病理学会, Japanese

胸腺プロテアソームの発現異常がT細胞分化に与える影響について　　　　　　　　　　　　　　　
小原 次郎, 外丸 詩野, 鈴木 小百合, 紺野 沙織, 村田 茂穂, 田中 啓二, 石津 明洋, 笠原 正典, 日本病理学会会誌, 101, 1, 435, 435, Mar. 2012
(一社)日本病理学会, Japanese

Expression of Thymoproteasome Subunit beta 5t in Type AB Thymoma
Y. Yamada, U. Tomaru, K. Kubota, A. Ishizu, T. Kiuchi, T. Mitsuhashi, Y. Matsuno, M. Kasahara, LABORATORY INVESTIGATION, 92, 493A, 493A, Feb. 2012
NATURE PUBLISHING GROUP, English, Summary international conference

Expression of Thymoproteasome Subunit beta 5t in Type AB Thymoma
Y. Yamada, U. Tomaru, K. Kubota, A. Ishizu, T. Kiuchi, T. Mitsuhashi, Y. Matsuno, M. Kasahara, MODERN PATHOLOGY, 25, 493A, 493A, Feb. 2012
NATURE PUBLISHING GROUP, English, Summary international conference

自己免疫の基礎病態 自己血管内皮細胞反応性血管炎惹起性T細胞の認識分子の同定(Identification of the vascular endothelial antigen recognized by T cells pathogenic for vasculitis)　　　　　　　　　　　　　　　
山口 まどか, 一條 加奈, 飯沼 千景, 脇 雅, 川上 愛, 佐々木 直美, 外丸 詩野, 笠原 正典, 石津 明洋, 日本免疫学会総会・学術集会記録, 40, 83, 83, Nov. 2011
(NPO)日本免疫学会, English

プロテアソーム活性の低下が腫瘍微小環境に与える影響 腫瘍増殖の抑制効果について(Proteasomal inhibition on the tumor microenvironment affects tumor proliferation)　　　　　　　　　　　　　　　
小野 綾子, 外丸 詩野, 石津 明洋, 小原 次郎, 紺野 沙織, 笠原 正典, 日本癌学会総会記事, 70回, 337, 337, Sep. 2011
日本癌学会, English

正常血圧性強皮症腎クリーゼの病理所見　　　　　　　　　　　　　　　
石津 明洋, 深谷 進司, 外丸 詩野, 古崎 章, 天崎 吉晴, 北海道医学雑誌, 86, 4-5, 241, 241, Aug. 2011
北海道医学会, Japanese

血管炎 自己血管内皮細胞反応性T細胞による血管炎発症モデル　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 55回・20回, 439, 439, Jun. 2011
(一社)日本リウマチ学会, Japanese

A model of vasculitis induced by autoreactive T cells against vascular endothelial cells
Akihiro Ishizu, Chihiro Iinuma, Masashi Waki, Madoka Yamaguchi, Naomi Sasaki, Utano Tomaru, Takashi Yoshiki, CLINICAL AND EXPERIMENTAL IMMUNOLOGY, 164, 128, 128, May 2011
WILEY-BLACKWELL, English, Summary international conference

プロピルチオウラシル(PTU)投与によるMPO-ANCA関連血管炎のモデル開発と病態解析　　　　　　　　　　　　　　　
中沢 大悟, 長谷川 梨沙, 一條 加奈, 西尾 妙織, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 100, 1, 315, 315, Mar. 2011
(一社)日本病理学会, Japanese

自己血管内皮細胞反応性T細胞による血管炎発症モデル　　　　　　　　　　　　　　　
脇 雅, 桜沢 貴代, 飯沼 千景, 山口 まどか, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 100, 1, 316, 316, Mar. 2011
(一社)日本病理学会, Japanese

プロテアソームの機能異常による病理作用 脂質代謝や脂肪肝との関連性について　　　　　　　　　　　　　　　
高橋 里美, 外丸 詩野, 合田 文, 小野 綾子, 小原 次郎, 宮武 由甲子, 村田 茂穂, 田中 啓二, 石津 明洋, 笠原 正典, 日本病理学会会誌, 100, 1, 333, 333, Mar. 2011
(一社)日本病理学会, Japanese

TNFα変換酵素の過剰発現による炎症・線維化病態への影響　　　　　　　　　　　　　　　
松井 由希, 深谷 進司, 外丸 詩野, 渥美 達也, 笠原 正典, 石津 明洋, 日本病理学会会誌, 100, 1, 344, 344, Mar. 2011
(一社)日本病理学会, Japanese

BCG接種後に判明した慢性肉芽腫症の一剖検例　　　　　　　　　　　　　　　
定本 圭弘, 大塚 紀幸, 藤田 裕美, 菊地 慶介, 外丸 詩野, 石津 明洋, 笠原 正典, 日本病理学会会誌, 100, 1, 494, 494, Mar. 2011
(一社)日本病理学会, Japanese

胸腺プロテアソームの異所性発現によるT細胞分化の異常　　　　　　　　　　　　　　　
小原 次郎, 外丸 詩野, 鈴木 小百合, 高橋 里美, 宮武 由甲子, 村田 茂穂, 田中 啓二, 石津 明洋, 笠原 正典, 日本病理学会会誌, 100, 1, 496, 496, Mar. 2011
(一社)日本病理学会, Japanese

von Recklinghausen病による動脈破裂の病理組織学的検討(第二報)　　　　　　　　　　　　　　　
木内 隆之, 高田 明生, 池田 仁, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 100, 1, 501, 501, Mar. 2011
(一社)日本病理学会, Japanese

Fcγ受容体を介したtrogocytosisの意義と制御機構の解析
益田紗季子, 岩崎沙理, 岩崎沙理, 佐藤樹里, 外丸詩野, 笠原正典, 石津明洋, 日本病理学会会誌, 100, 1, 2011

Circulating fibrocyteの性質異常とケロイド病態との関連性 創傷モデルマウスを用いた検討　　　　　　　　　　　　　　　
長尾 宗朝, 小山 明彦, 村尾 尚規, 小浦場 祥夫, 古川 洋志, 山本 有平, 外丸 詩野, 石津 明洋, 日本形成外科学会会誌, 30, 11, 649, 649, Nov. 2010
(一社)日本形成外科学会, Japanese

【血管炎症候群の新展開】 トランスクリプトミクスによる血管炎関連因子の探索　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 村井 太一, 山本 智宏, 吉木 敬, 炎症と免疫, 18, 5, 509, 513, Aug. 2010
JMAAVは、わが国におけるMPO-ANCA関連血管炎の標準的治療プロトコール策定を目的として設立された多施設共同研究組織である。われわれは、MPO-ANCA関連血管炎の病因・病態を明らかにするために、JMAAVに登録された症例の治療前および治療開始1週間後の末梢血をサンプルとして、網羅的遺伝子発現解析(トランスクリプトーム解析)をおこなった。治療後に寛解し、その後も寛解が維持された症例において、88個の遺伝子が治療前後で統計学的に有意な発現変化を示した。このうち、治療後に発現が減少した遺伝子は66個、治療後に発現が増加した遺伝子は22個であった。これらのなかから、治療開始後の早期にMPO-ANCA関連血管炎の治療後の予後を予測する16個の遺伝子を抽出した。(著者抄録), (株)先端医学社, Japanese

好酸球性肉芽腫性血管炎により小腸壊死を呈したが、アレルギー症状や好酸球増加を認めず、診断に苦慮した症例　　　　　　　　　　　　　　　
石津 明洋, 高橋 利幸, 外丸 詩野, 深谷 進司, 堀田 彰一, 森田 高行, 古崎 章, 天崎 吉晴, 北海道医学雑誌, 85, 4, 281, 281, Jul. 2010
北海道医学会, Japanese

動脈破裂をきたしたvon Recklinghausen病の2症例 破裂血管におけるhypoxia-inducible factor-1a発現の検討　　　　　　　　　　　　　　　
木内 隆之, 外丸 詩野, 高田 明生, 武藤 修一, 大岡 智学, 村上 達哉, 宮武 司, 大場 淳一, 青木 秀俊, 石津 明洋, 脈管学, 50, 2, 220, 221, Apr. 2010
(一社)日本脈管学会, Japanese

OP-052-5 発現頻度・発現パターンから見た大腸癌における癌幹細胞マーカーCD133の臨床的意義についての検討(腫瘍基礎-2,一般口演,第110回日本外科学会定期学術集会)
高橋 周作, 神山 俊哉, 島田 信吾, 工藤 岳秋, 廣瀬 邦弘, 佐治 裕, 崎浜 秀康, 尾崎 倫孝, 外丸 詩野, 石津 明洋, 藤堂 省, 日本外科学会雑誌, 111, 2, 372, 372, 05 Mar. 2010
一般社団法人日本外科学会, Japanese

血管炎・APS 血管炎惹起性T細胞クローンの樹立と解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 54回・19回, 543, 543, Mar. 2010
(一社)日本リウマチ学会, Japanese

TNFα変換酵素(TACE)トランスジェニックマウスの解析　　　　　　　　　　　　　　　
深谷 進司, 外丸 詩野, 松井 由希, 保田 晋助, 堀田 哲也, 片岡 浩, 渥美 達也, 小池 隆夫, 笠原 正典, 石津 明洋, 日本病理学会会誌, 99, 1, 206, 206, Mar. 2010
(一社)日本病理学会, Japanese

血管炎惹起性T細胞クローンの樹立と解析　　　　　　　　　　　　　　　
飯沼 千景, 脇 雅, 山口 まどか, 佐々木 直美, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 99, 1, 210, 210, Mar. 2010
(一社)日本病理学会, Japanese

胸腺プロテアソームの異所性発現によるT細胞分化の異常　　　　　　　　　　　　　　　
鈴木 小百合, 外丸 詩野, 高橋 里実, 小原 次郎, 風巻 拓, 小野 綾子, 宮武 由甲子, 村田 茂穂, 田中 啓二, 石津 明洋, 笠原 正典, 日本病理学会会誌, 99, 1, 217, 217, Mar. 2010
(一社)日本病理学会, Japanese

プロテアソームの機能異常による病理作用　　　　　　　　　　　　　　　
高橋 里実, 外丸 詩野, 鈴木 小百合, 小野 綾子, 宮武 由甲子, 風巻 拓, 小原 次郎, 村田 茂穂, 田中 啓二, 石津 明洋, 笠原 正典, 日本病理学会会誌, 99, 1, 225, 225, Mar. 2010
(一社)日本病理学会, Japanese

von Recklinghausen病による動脈破裂の病理組織学的検討　　　　　　　　　　　　　　　
木内 隆之, 外丸 詩野, 高田 明生, 石津 明洋, 日本病理学会会誌, 99, 1, 367, 367, Mar. 2010
(一社)日本病理学会, Japanese

血管炎・APS JMAAVトランスクリプトーム解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 吉木 敬, 湯村 和子, 山縣 邦弘, 山田 秀裕, 熊谷 俊一, 黒川 真奈絵, 須賀 万智, 尾崎 承一, Japanese study, group for, MPO-ANCA-associated vasculitis(JMAAV, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 54回・19回, 543, 543, Mar. 2010
(一社)日本リウマチ学会, Japanese

ヒト末梢血に検出されるCD8陽性単球・顆粒球の解析
益田紗季子, 岩崎沙理, 岩崎沙理, 馬場智久, 勝俣一晃, 外丸詩野, 笠原正典, 石津明洋, 日本病理学会会誌, 99, 1, 2010

MPO-ANCA陽性であったが糸球体腎炎や血管炎は確認されず、間質性肺炎に感染症を合併して死亡した症例
石津 明洋, 岩崎 沙理, 外丸 詩野, 武田 広子, 大塚 紀幸, 富居 一範, 笠原 正典, 清水 健一, 南須原 康行, 西村 正治, 北海道医学雑誌, 84, 5, 403, 403, Sep. 2009
北海道医学会, Japanese

当科で経験した耳下腺導管癌の1例　　　　　　　　　　　　　　　
前田 昌紀, 黒田 徹, 富居 一範, 外丸 詩野, 石津 明洋, 耳鼻咽喉科臨床 補冊, 補冊124, 134, 134, Jun. 2009
耳鼻咽喉科臨床学会, Japanese

血管炎 MPO-ANCA関連血管炎患者末梢血のトランスクリプトーム解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 岩崎 沙理, 吉木 敬, 尾崎 承一, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 53回・18回, 209, 209, Mar. 2009
(一社)日本リウマチ学会, Japanese

MPO-ANCA関連血管炎患者末梢血のトランスクリプトーム解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 岩崎 沙理, 飯沼 千景, 村井 太一, 山本 智宏, 吉木 敬, 尾崎 承一, 日本病理学会会誌, 98, 1, 211, 211, Mar. 2009
(一社)日本病理学会, Japanese

胸腺プロテアソームの発現とT細胞分化に関する検討　　　　　　　　　　　　　　　
鈴木 小百合, 外丸 詩野, 石津 明洋, 高橋 里実, 小原 次郎, 風巻 拓, 宮武 由甲子, 村田 茂穂, 田中 啓二, 笠原 正典, 日本病理学会会誌, 98, 1, 277, 277, Mar. 2009
(一社)日本病理学会, Japanese

自己血管内皮細胞反応性ラットT細胞の解析　　　　　　　　　　　　　　　
飯沼 千景, 佐々木 直美, 岩崎 沙理, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 98, 1, 292, 292, Mar. 2009
(一社)日本病理学会, Japanese

プロテアソームのキモトリプシン様活性の異常と病態形成に関する検討　　　　　　　　　　　　　　　
外丸 詩野, 石津 明洋, 高橋 里実, 鈴木 小百合, 小原 次郎, 風巻 拓, 宮武 由甲子, 村田 茂穂, 田中 啓二, 笠原 正典, 日本病理学会会誌, 98, 1, 368, 368, Mar. 2009
(一社)日本病理学会, Japanese

分子標的薬sunitinib投与症例における甲状腺萎縮の病理学的解析　　　　　　　　　　　　　　　
中村 静香, 大塚 紀幸, 鈴木 昭, 富居 一範, 外丸 詩野, 笠原 正典, 石津 明洋, 日本病理学会会誌, 98, 1, 400, 400, Mar. 2009
(一社)日本病理学会, Japanese

【中小型血管炎の新展開】 MPO-ANCA関連血管炎の予後予測因子の探索　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 村井 太一, 吉木 敬, 尾崎 承一, 脈管学, 49, 1, 45, 51, Feb. 2009
(一社)日本脈管学会, Japanese

ヒト末梢血に検出されるCD8陽性単球の解析
岩崎沙理, 岩崎沙理, 馬場智久, 益田紗季子, 勝俣一晃, 外丸詩野, 笠原正典, 石津明洋, 日本病理学会会誌, 98, 1, 2009

ヒト末梢血におけるCD8陽性単球の解析
岩崎沙理, 益田紗季子, 馬場智久, 勝俣一晃, 外丸詩野, 笠原正典, 石津明洋, 臨床病理, 57, 2009

リツキシマブ投与後に日和見感染症を併発して死亡した顕微鏡的多発血管炎の一剖検例
外丸 詩野, 富居 一範, 武田 広子, 小川 弥生, 堀田 哲也, 深谷 進司, 橋本 陶子, 石津 明洋, 北海道医学雑誌, 84, 1, 62, 62, Jan. 2009
北海道医学会, Japanese

潰瘍性大腸炎における末梢血および大腸粘膜内NKG2A+T細胞の減少　　　　　　　　　　　　　　　
桂田 武彦, 外丸 詩野, 笠原 正典, 馬場 智久, 古川 滋, 武田 宏司, 浅香 正博, 田中 淳司, 石津 明洋, 北海道医学雑誌, 84, 1, 62, 62, Jan. 2009
北海道医学会, Japanese

MPO-ANCA関連血管炎患者末梢血のトランスクリプトーム解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 村井 太一, 西平 順, 吉木 敬, 尾崎 承一, 脈管学, 48, 5, 489, 490, Oct. 2008
(一社)日本脈管学会, Japanese

顕微鏡的多発血管炎の肺病変について　　　　　　　　　　　　　　　
斉藤 永秀, 岩崎 沙理, 外丸 詩野, 石津 明洋, 脈管学, 48, 5, 496, 496, Oct. 2008
(一社)日本脈管学会, Japanese

リツキシマブ投与後に日和見感染症を併発して死亡した顕微鏡的多発血管炎の一剖検例
外丸 詩野, 武田 広子, 小川 弥生, 堀田 哲也, 深谷 進司, 橋本 陶子, 富居 一範, 石津 明洋, 脈管学, 48, 5, 494, 494, Oct. 2008
(一社)日本脈管学会, Japanese

顕微鏡的多発血管炎患者末梢血のトランスクリプトーム解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 村井 太一, 吉木 敬, 尾崎 承一, 日本病理学会会誌, 97, 2, 23, 23, Sep. 2008
(一社)日本病理学会, Japanese

肺限局型MPO-ANCA関連血管炎と考えられた一剖検例　　　　　　　　　　　　　　　
外丸 詩野, 武田 広子, 岩崎 沙理, 大塚 紀幸, 富居 一範, 笠原 正典, 清水 健一, 南須原 康行, 西村 正治, 石津 明洋, 日本病理学会会誌, 97, 2, 33, 33, Sep. 2008
(一社)日本病理学会, Japanese

新規NKG2DリガンドH60b、H60cの機能解析(Functional analysis of novel members of NKG2D ligand, H60b and H60c)　　　　　　　　　　　　　　　
富居 一範, 吉田 繁, 大塚 紀幸, 外丸 詩野, 笠原 正典, 日本癌学会総会記事, 67回, 443, 444, Sep. 2008
日本癌学会, English

血管炎症候群研究の進歩 自己血管内皮細胞反応性ラットT細胞の解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 飯沼 千景, 岩崎 沙理, 佐々木 直美, 馬場 智久, 笠原 正典, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 52回・17回, 156, 156, Apr. 2008
(一社)日本リウマチ学会, Japanese

潰瘍性大腸炎におけるNKG2A陽性T細胞の減少と病態への関与　　　　　　　　　　　　　　　
外丸 詩野, 石津 明洋, 笠原 正典, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 52回・17回, 280, 280, Apr. 2008
(一社)日本リウマチ学会, Japanese

炎症研究の新しい展開 CD4/CD8 double positiveマクロファージの発見と自然免疫系における新しいエフェクター細胞としての展開　　　　　　　　　　　　　　　
馬場 智久, 岩崎 浩理, 外丸 詩野, 池田 仁, 吉木 敬, 笠原 正典, 向田 直史, 石津 明洋, 日本病理学会会誌, 97, 1, 142, 142, Mar. 2008
(一社)日本病理学会, Japanese

自己免疫病発症機構の新たな概念 自己血管内皮細胞反応性ラットT細胞の解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 岩崎 沙理, 飯沼 千景, 佐藤 亜矢, 佐々木 直美, 馬場 智久, 笠原 正典, 吉木 敬, 日本病理学会会誌, 97, 1, 169, 169, Mar. 2008
(一社)日本病理学会, Japanese

胸腺プロテアソームの発現とT細胞分化に関する検討　　　　　　　　　　　　　　　
外丸 詩野, 石津 明洋, 宮武 由甲子, 高橋 里実, 小原 次郎, 村田 茂穂, 田中 啓二, 笠原 正典, 日本病理学会会誌, 97, 1, 203, 203, Mar. 2008
(一社)日本病理学会, Japanese

ヒト末梢血におけるCD8陽性単球の解析　　　　　　　　　　　　　　　
岩崎 沙理, 馬場 智久, 勝俣 一晃, 外丸 詩野, 笠原 正典, 石津 明洋, 日本病理学会会誌, 97, 1, 203, 203, Mar. 2008
(一社)日本病理学会, Japanese

MPO-ANCA関連血管炎患者末梢血のトランスクリプトーム解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 村井 太一, 西平 順, 吉木 敬, 尾崎 承一, 日本病理学会会誌, 97, 1, 261, 261, Mar. 2008
(一社)日本病理学会, Japanese

ヒト組織におけるプロテアソームサブユニットβ5t発現に関する検討　　　　　　　　　　　　　　　
外丸 詩野, 石津 明洋, 宮武 由甲子, 鈴木 小百合, 風巻 拓, 村田 茂穂, 田中 啓二, 笠原 正典, 日本病理学会会誌, 97, 1, 309, 309, Mar. 2008
(一社)日本病理学会, Japanese

ヒト組織におけるプロテアソームサブユニットβ5t発現に関する検討　　　　　　　　　　　　　　　
風巻 拓, 外丸 詩野, 石津 明洋, 鈴木 小百合, 笠原 正典, 日本病理学会会誌, 97, 1, 392, 392, Mar. 2008
(一社)日本病理学会, Japanese

潰瘍性大腸炎における末梢血および大腸粘膜内NKG2A+T細胞の減少　　　　　　　　　　　　　　　
桂田 武彦, 外丸 詩野, 馬場 智久, 古川 滋, 武田 宏司, 田中 淳司, 浅香 正博, 石津 明洋, 笠原 正典, 日本病理学会会誌, 97, 1, 233, 233, Mar. 2008
(一社)日本病理学会, Japanese

NKG2DリガンドH60b、H60cの機能解析　　　　　　　　　　　　　　　
吉田 繁, 富居 一範, 大塚 紀幸, 外丸 詩野, 笠原 正典, 日本病理学会会誌, 97, 1, 203, 203, Mar. 2008
(一社)日本病理学会, Japanese

Churg-Strauss syndrome(CSS)-associated eosinophilic endocarditisの一例　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 平林 鑑, 三山 博史, 筒井 裕之, 保田 晋介, 吉木 敬, 北海道医学雑誌, 82, 6, 441, 442, Nov. 2007
北海道医学会, Japanese

潰瘍性大腸炎における末梢血中NKG2A陽性T細胞の減少と病態への関与　　　　　　　　　　　　　　　
桂田 武彦, 外丸 詩野, 鈴木 昭, 笠原 正典, 古川 滋, 武田 宏司, 浅香 正博, 田中 淳司, 石津 明洋, 北海道医学雑誌, 82, 6, 441, 441, Nov. 2007
北海道医学会, Japanese

抗腫瘍免疫反応の免疫調節 CD4+/CD8+マクロファージはNKG2D及びグランザイム/パーフォリン依存性メカニズムを介して殺腫瘍細胞作用を示す(Immunoregulation of Anti-tumor Immune Responses CD4+/CD8+ macrophages kill tumor cells through an NKG2D-and granzyme/perforin-dependent mechanism)　　　　　　　　　　　　　　　
石津 明洋, 馬場 智久, 岩崎 沙理, 外丸 詩野, 笠原 正典, 吉木 敬, 日本癌学会総会記事, 66回, 74, 74, Aug. 2007
日本癌学会, English

マウスH60ファミリーの異なる発現パターンとNKG2Dへの結合親和性を持つ二つの新しいNKG2Dリガンド(Two Novel NKG2D Ligands of the Mouse H60 Family with Differential Expression Patterns and Binding Affinities to NKG2D)　　　　　　　　　　　　　　　
吉田 繁, 高田 明生, 富居 一範, 宮武 由甲子, 外丸 詩野, 笠原 正典, 日本癌学会総会記事, 66回, 41, 41, Aug. 2007
日本癌学会, English

潰瘍性大腸炎における末梢血中NKG2A陽性T細胞の減少と病態への関与　　　　　　　　　　　　　　　
外丸 詩野, 石津 明洋, 笠原 正典, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 51回・16回, 338, 338, Apr. 2007
(一社)日本リウマチ学会, Japanese

自己血管内皮細胞反応性ラットT細胞の解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 岩崎 沙理, 笠原 正典, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 51回・16回, 373, 373, Apr. 2007
(一社)日本リウマチ学会, Japanese

CD4/CD8 double positiveマクロファージによる抗腫瘍メカニズムの解明　　　　　　　　　　　　　　　
馬場 智久, 岩崎 沙理, 石津 明洋, 鈴木 昭, 外丸 詩野, 池田 仁, 吉木 敬, 笠原 正典, 日本病理学会会誌, 96, 1, 205, 205, Feb. 2007
(一社)日本病理学会, Japanese

自己血管内皮細胞反応性ラットT細胞の解析　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 馬場 智久, 佐々木 直美, 岩崎 沙理, 笠原 正典, 吉木 敬, 日本病理学会会誌, 96, 1, 222, 222, Feb. 2007
(一社)日本病理学会, Japanese

潰瘍性大腸炎における末梢血中NKG2A陽性T細胞の減少と病態への関与　　　　　　　　　　　　　　　
桂田 武彦, 外丸 詩野, 古川 滋, 武田 宏司, 田中 淳司, 浅香 正博, 鈴木 昭, 石津 明洋, 笠原 正典, 日本病理学会会誌, 96, 1, 226, 226, Feb. 2007
(一社)日本病理学会, Japanese

新しいマウスNKG2Dリガンドの同定と解析　　　　　　　　　　　　　　　
吉田 繁, 富居 一範, 高田 明生, 宮武 由甲子, 鈴木 昭, 石津 明洋, 外丸 詩野, 笠原 正典, 日本病理学会会誌, 96, 1, 205, 205, Feb. 2007
(一社)日本病理学会, Japanese

生殖細胞特異的に発現する新規MHC class I様分子の解析　　　　　　　　　　　　　　　
富居 一範, 吉田 繁, 宮武 由甲子, 高田 明生, 鈴木 昭, 石津 明洋, 外丸 詩野, 笠原 正典, 日本病理学会会誌, 96, 1, 206, 206, Feb. 2007
(一社)日本病理学会, Japanese

Micronodular thymoma with lymphoid hyperplasia(MNT)の一例　　　　　　　　　　　　　　　
古西 崇寛, 山田 綾子, 富居 一範, 鈴木 昭, 外丸 詩野, 石津 明洋, 笠原 正典, 日本病理学会会誌, 96, 1, 355, 355, Feb. 2007
(一社)日本病理学会, Japanese

メソトレキセート(MTX)関連リンパ増殖性疾患と考えられた症例　　　　　　　　　　　　　　　
石津 明洋, 外丸 詩野, 鈴木 昭, 金井 直樹, 北海道医学雑誌, 81, 6, 562, 562, Nov. 2006
北海道医学会, Japanese

NK細胞とMHC受容体 新たなマウスNKG2D ligand様分子の同定とその機能に関する解析　　　　　　　　　　　　　　　
富居 一範, 吉田 繁, 宮武 由甲子, 鈴木 昭, 石津 明洋, 外丸 詩野, 笠原 正典, 日本免疫学会総会・学術集会記録, 36, 253, 253, Nov. 2006
(NPO)日本免疫学会, Japanese

CD4/CD8 double positiveマクロファージの抗腫瘍メカニズムの検討　　　　　　　　　　　　　　　
馬場 智久, 岩崎 沙理, 石津 明洋, 外丸 詩野, 鈴木 昭, 池田 仁, 吉木 敬, 笠原 正典, 日本癌学会総会記事, 65回, 329, 329, Sep. 2006
日本癌学会, Japanese

CD4/CD8 double positiveマクロファージの抗腫瘍活性の検討　　　　　　　　　　　　　　　
馬場 智久, 岩崎 沙理, 石津 明洋, 鈴木 昭, 外丸 詩野, 池田 仁, 吉木 敬, 笠原 正典, 日本病理学会会誌, 95, 1, 201, 201, Apr. 2006
(一社)日本病理学会, Japanese

血管炎惹起性ラットT細胞の樹立と解析　　　　　　　　　　　　　　　
佐々木 直美, 石津 明洋, 外丸 詩野, 鈴木 昭, 笠原 正典, 吉木 敬, 日本病理学会会誌, 95, 1, 228, 228, Apr. 2006
(一社)日本病理学会, Japanese

HTLV-I持続感染ラットの脊髄症発症におけるNeuronal IFN-γの役割　　　　　　　　　　　　　　　
宮武 由甲子, 石津 明洋, 池田 仁, 馬場 智久, 鈴木 昭, 外丸 詩野, 笠原 正典, 吉木 敬, 日本病理学会会誌, 95, 1, 231, 231, Apr. 2006
(一社)日本病理学会, Japanese

子宮頸部病理検体において検出されるHPVジェノタイプと臨床病理学的因子の関連　　　　　　　　　　　　　　　
道又 理恵, 尾川 直樹, 石津 明洋, 外丸 詩野, 鈴木 昭, 笠原 正典, 吉木 敬, 日本病理学会会誌, 95, 1, 291, 291, Apr. 2006
(一社)日本病理学会, Japanese

リバビリン併用インターフェロン療法開始後,急速に肝不全をきたして死亡したHCVとHIVの重複感染例　　　　　　　　　　　　　　　
三次 有奈, 浅野 拓行, 石津 明洋, 外丸 詩野, 鈴木 昭, 池田 仁, 笠原 正典, 日本病理学会会誌, 95, 1, 382, 382, Apr. 2006
(一社)日本病理学会, Japanese

Granulomatous slack skinの一例　　　　　　　　　　　　　　　
石井 生, 久田 敦史, 鈴木 昭, 近藤 信夫, 外丸 詩野, 石津 明洋, 笠原 正典, 日本病理学会会誌, 95, 1, 385, 385, Apr. 2006
(一社)日本病理学会, Japanese

潰瘍性大腸炎患者の末梢血Tリンパ球における抑制性NKレセプター発現の検討　　　　　　　　　　　　　　　
桂田 武彦, 外丸 詩野, 鈴木 昭, 石津 明洋, 古川 滋, 武田 宏司, 田中 淳司, 浅香 正博, 笠原 正典, 日本病理学会会誌, 95, 1, 196, 196, Apr. 2006
(一社)日本病理学会, Japanese

動物モデルでの自己免疫疾患の解析 血管炎惹起性ラットT細胞の樹立と解析　　　　　　　　　　　　　　　
石津 明洋, 佐々木 直美, 外丸 詩野, 笠原 正典, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 50回・15回, 160, 160, Mar. 2006
(一社)日本リウマチ学会, Japanese

自己免疫状態におけるヒト内在性レトロウイルスHERV-Rの抗原性について　　　　　　　　　　　　　　　
石津 明洋, 佐々木 直美, 田中 敏, 大塚 紀幸, 鈴木 昭, 外丸 詩野, 池田 仁, 笠原 正典, 吉木 敬, 日本免疫学会総会・学術集会記録, 35, 35, 35, Nov. 2005
(NPO)日本免疫学会, Japanese

細胞傷害活性をもつCD4/CD8 double positiveマクロファージの解析　　　　　　　　　　　　　　　
馬場 智久, 石津 明洋, 外丸 詩野, 鈴木 昭, 池田 仁, 吉木 敬, 笠原 正典, 日本免疫学会総会・学術集会記録, 35, 124, 124, Nov. 2005
(NPO)日本免疫学会, Japanese

HTLV-I感染ラットにおける脊髄症の発症にはニューロンでのIL-12を介したIFN-γの産生不応答が関与する　　　　　　　　　　　　　　　
宮武 由甲子, 池田 仁, 石津 明洋, 外丸 詩野, 鈴木 昭, 吉木 敬, 笠原 正典, 日本免疫学会総会・学術集会記録, 35, 177, 177, Nov. 2005
(NPO)日本免疫学会, Japanese

原疾患の増悪に対するステロイド増量後に大動脈瘤破裂をきたして死亡した皮膚筋炎の症例　　　　　　　　　　　　　　　
佐々木 直美, 石津 明洋, 鈴木 昭, 外丸 詩野, 笠原 正典, 保田 晋助, 桜井 典之, 北海道医学雑誌, 80, 5, 537, 537, Sep. 2005
北海道医学会, Japanese

HTLV-I感染によるHAMラット病の発症はニューロンにおけるIL-12を介したIFN-γの反応性に依存する　　　　　　　　　　　　　　　
宮武 由甲子, 池田 仁, 馬場 智久, 道又 理恵, 鈴木 昭, 外丸 詩野, 石津 明洋, 吉木 敬, 笠原 正典, 日本癌学会総会記事, 64回, 343, 343, Sep. 2005
日本癌学会, Japanese

DNAアレイ法を用いた子宮頸癌症例のHPVジェノタイピング　　　　　　　　　　　　　　　
道又 理恵, 尾川 直樹, 石津 明洋, 渡利 英道, 鈴木 昭, 外丸 詩野, 櫻木 範明, 笠原 正典, 吉木 敬, 日本癌学会総会記事, 64回, 93, 93, Sep. 2005
日本癌学会, Japanese

膠原病研究の新たな展開 分子病態と制御モデル HTLV-I pX遺伝子導入ラットと正常Wistarラットの交配種における胸腺異常とchronic GVHD様疾患の発症　　　　　　　　　　　　　　　
石津 明洋, 馬場 智久, 外丸 詩野, 池田 仁, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 49回・14回, 116, 116, Apr. 2005
(一社)日本リウマチ学会, Japanese

ラットモデルを用いた壊死性血管炎発症機序の解析　　　　　　　　　　　　　　　
佐々木 直美, 石津 明洋, 鈴木 昭, 外丸 詩野, 笠原 正典, 吉木 敬, 日本病理学会会誌, 94, 1, 213, 213, Mar. 2005
(一社)日本病理学会, Japanese

HTLV-I pX遺伝子導入ラットと正常Wistarラットの交配種における胸腺異常と免疫病理　　　　　　　　　　　　　　　
馬場 智久, 石津 明洋, 鈴木 昭, 外丸 詩野, 池田 仁, 笠原 正典, 吉木 敬, 日本病理学会会誌, 94, 1, 219, 219, Mar. 2005
(一社)日本病理学会, Japanese

ヒトCD4,CCR5,MHC class II transactivator遺伝子導入ラットの作製と末梢血単核球へのHIV-1の感染　　　　　　　　　　　　　　　
鈴木 昭, 陳 晶, 馬場 智久, 外丸 詩野, 石津 明洋, 池田 仁, 笠原 正典, 吉木 敬, 日本病理学会会誌, 94, 1, 225, 225, Mar. 2005
(一社)日本病理学会, Japanese

T細胞による抗原提示と免疫制御　　　　　　　　　　　　　　　
外丸 詩野, 山野 嘉久, 鈴木 昭, 石津 明洋, 吉木 敬, 笠原 正典, 日本病理学会会誌, 94, 1, 370, 370, Mar. 2005
(一社)日本病理学会, Japanese

HTLV-I感染によるHAMラット病感受性および抵抗性は脊髄における宿主の対ウイルス反応性に依存する　　　　　　　　　　　　　　　
宮武 由甲子, 池田 仁, 鈴木 昭, 馬場 智久, 道又 理恵, 外丸 詩野, 石津 明洋, 吉木 敬, 日本免疫学会総会・学術集会記録, 34, 95, 95, Nov. 2004
(NPO)日本免疫学会, Japanese

T細胞による抗原提示と免疫制御機構　　　　　　　　　　　　　　　
外丸 詩野, 山野 嘉久, 石津 明洋, 吉木 敬, 日本免疫学会総会・学術集会記録, 34, 100, 100, Nov. 2004
(NPO)日本免疫学会, Japanese

HTLV-IpX遺伝子導入ラットと正常Wistarラットの交配種における胸腺異常とchronic GVHD様疾患の発症　　　　　　　　　　　　　　　
馬場 智久, 石津 明洋, 宮武 由甲子, 鈴木 昭, 外丸 詩野, 池田 仁, 吉木 敬, 日本免疫学会総会・学術集会記録, 34, 278, 278, Nov. 2004
(NPO)日本免疫学会, Japanese

ペプチド/HLA複合体の抗原特異的T細胞による獲得 抗原提示の新しい増幅メカニズム　　　　　　　　　　　　　　　
外丸 詩野, 山野 嘉久, 石津 明洋, 池田 仁, 吉木 敬, 日本病理学会会誌, 93, 1, 239, 239, May 2004
(一社)日本病理学会, Japanese

HTLV-1 pXトランスジェニックラットと正常Wistarラットの交配種における全身性炎症性疾患の発症　　　　　　　　　　　　　　　
馬場 智久, 石津 明洋, 宮武 由甲子, 外丸 詩野, 池田 仁, 吉木 敬, 日本病理学会会誌, 93, 1, 240, 240, May 2004
(一社)日本病理学会, Japanese

HTLV-I感染によるHAM感受性ラット及び抵抗性ラットの脊髄における遺伝子発現の解析　　　　　　　　　　　　　　　
宮武 由甲子, 外丸 詩野, 石津 明洋, 池田 仁, 吉木 敬, 日本病理学会会誌, 93, 1, 249, 249, May 2004
(一社)日本病理学会, Japanese

HTLV-IトランスジェニックラットにおけるCD4+CD25+regulatory T cellの解析　　　　　　　　　　　　　　　
早瀬 広子, 石津 明洋, 宮武 由甲子, 外丸 詩野, 池田 仁, 吉木 敬, 日本病理学会会誌, 93, 1, 294, 294, May 2004
(一社)日本病理学会, Japanese

ヒト大腸癌培養株DLD-1細胞とその5-FU耐性株の比較　　　　　　　　　　　　　　　
伊藤 僚子, 吉木 敬, 池田 仁, 石津 明洋, 外丸 詩野, 日本病理学会会誌, 93, 1, 304, 304, May 2004
(一社)日本病理学会, Japanese

免疫の多様性 自己免疫疾患モデルとしてのHTLV-I env-pX遺伝子導入ラット　　　　　　　　　　　　　　　
石津 明洋, 早瀬 広子, 宮武 由甲子, 富居 一範, 樋口 正人, 阿部 麻美, 外丸 詩野, 池田 仁, 吉木 敬, 日本病理学会会誌, 93, 1, 163, 163, May 2004
(一社)日本病理学会, Japanese

リウマチ性疾患を発症するHTLV-I遺伝子導入ラットにおける免疫制御性CD25+4+T細胞の解析　　　　　　　　　　　　　　　
早瀬 広子, 石津 明洋, 宮武 由甲子, 外丸 詩野, 池田 仁, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 48回, 172, 172, Mar. 2004
(一社)日本リウマチ学会, Japanese

転写因子CREBを標的とした関節リウマチの遺伝子治療モデル　　　　　　　　　　　　　　　
石津 明洋, 馬場 智久, 早瀬 広子, 外丸 詩野, 池田 仁, 吉木 敬, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 48回, 317, 317, Mar. 2004
(一社)日本リウマチ学会, Japanese

HTLV-1トランスジェニックラットにおけるCD25+4+ regulatory T cellの解析　　　　　　　　　　　　　　　
早瀬 広子, 石津 明洋, 宮武 由甲子, 外丸 詩野, 池田 仁, 吉木 敬, 日本免疫学会総会・学術集会記録, 33, 42, 42, Nov. 2003
(NPO)日本免疫学会, Japanese

HTLV-1 pXトランスジェニックラット胸腺腫の悪性化とp16遺伝子異常の解析　　　　　　　　　　　　　　　
辻 隆裕, 池田 仁, 外丸 詩野, 石津 明洋, 吉木 敬, 日本癌学会総会記事, 62回, 241, 241, Aug. 2003
日本癌学会, Japanese

HTLV-1 Env領域におけるCD8+T細胞エピトープの同定 抗原特異的Tリンパ球の新たな検出法　　　　　　　　　　　　　　　
外丸 詩野, 山野 嘉久, 石津 明洋, 池田 仁, 吉木 敬, 日本癌学会総会記事, 62回, 242, 242, Aug. 2003
日本癌学会, Japanese

Detection of HTLV-I Tax11-19 peptide/HLA-A*201 complexes are overexpressed in HAM/TSP patients.
Y Yamano, CJ Cohen, U Tomaru, Y Reiter, S Jacobson, AIDS RESEARCH AND HUMAN RETROVIRUSES, 19, S38, S38, 2003
MARY ANN LIEBERT INC PUBL, English, Summary international conference

Components of the cellular membrane involved in HTLV-I Env-mediated entry.
KS Jones, U Tomaru, Y Yamano, C Sadowski, M Dambach, S Jacobson, FW Ruscetti, AIDS RESEARCH AND HUMAN RETROVIRUSES, 19, S28, S28, 2003
MARY ANN LIEBERT INC PUBL, English, Summary international conference

Transference of peptide/HLA complexes among HTLV-I-specific T cells; A new concept of antigen spread in HTLV-I-associated disease.
U Tomaru, Y Yamano, M Nagai, S Jacobson, AIDS RESEARCH AND HUMAN RETROVIRUSES, 19, S8, S9, 2003
MARY ANN LIEBERT INC PUBL, English, Summary international conference

Detection of HTLV-I-specific T cells and novel CD8+ T cell epitopes by acquisition of peptide/HLA-GFP complexes.
U Tomaru, Y Yamano, M Nagai, D Maric, PT Kaumaya, W Biddison, S Jacobson, AIDS RESEARCH AND HUMAN RETROVIRUSES, 19, S44, S45, 2003
MARY ANN LIEBERT INC PUBL, English, Summary international conference

T cell differentiation phenotype and cytolytic function of CMV and HTLV-I-specific CD8+ T cells in HAM/TSP patients: Evidence for insufficient control of persistent HTLV-I infection.
Y Yamano, U Tomaru, M Nagai, S Jacobson, AIDS RESEARCH AND HUMAN RETROVIRUSES, 19, S41, S41, 2003
MARY ANN LIEBERT INC PUBL, English, Summary international conference

Blood levels of vascular endothelial growth factor in obstructive sleep apneahypopnea syndrome [5] (multiple letters)
Yoshihisa Yamano, Masahiro Nagai, Meghan Brennan, Carlos A. Mora, Samantha S. Soldan, Utano Tomaru, Norihiro Takenouchi, Shuji Izumo, Mitsuhiro Osame, Steven Jacobson, Blood, 99, 1, 393, 394, 01 Jan. 2002
English, Report scientific journal

Real-time quantitative RT-PCR analysis of human T-cell lymphotropic virus type-1 (HTLV-I) tax mRNA: Correlation with HTLV-I proviral DNA load, Tax protein expression, Tax specific CD8+T-cell frequency and disease severity in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP)
Y Yamano, M Nagai, MG Brennan, CA Mora, SS Soldan, U Tomaru, M Osame, S Jacobson, AIDS RESEARCH AND HUMAN RETROVIRUSES, 17, S25, S25, 2001
MARY ANN LIEBERT INC PUBL, English, Summary international conference

Human T lymphocyte virus type I-induced myeloneuropathy in rats: Implication of local activation of the pX and tumor necrosis factor-alpha genes in pathogenesis
U Tomaru, H Ikeda, O Ohya, M Abe, T Kasai, Yamasita, I, K Morita, A Wakisaka, T Yoshiki, JOURNAL OF INFECTIOUS DISEASES, 174, 2, 318, 323, Aug. 1996
The pathogenetic roles of human T lymphocyte virus type I (HTLV-I) and cytokines were investigated in HTLV-I-induced myeloneuropathy in Wistar-King-Aptekman-Hokudai rats. In the nervous system, pX messenger RNAs of HTLV-I were selectively expressed in the diseased spinal cord and peripheral nerves but not in the unaffected cerebrum and cerebellum, even though proviral DNAs were consistently identified in these tissues. Among several cytokines examined, mRNA expression and production of tumor necrosis factor (TNF)-alpha in the spinal cord and cerebrospinal fluid correlated positively with the development of spinal cord lesions. The collective evidence strongly suggests that selective activation of HTLV-I, in particular Tax expression and production of TNF-alpha induced by HTLV-I infection in target spinal cord and peripheral nerves, is causally related to apoptotic death of oligodendrocytes and Schwann cells, a major pathogenetic pathway of the HTLV-I-induced myeloneuropathy., UNIV CHICAGO PRESS, English

HTLV-I INDUCED MYELONEUROPATHY IN WKAH RATS - APOPTOSIS AND LOCAL ACTIVATION OF THE HTLV-I PX AND TNF-ALPHA GENES IMPLICATED IN THE PATHOGENESIS
U TOMARU, H IKEDA, O OHYA, T KASAI, YAMASHITA, I, K MORITA, A WAKISAKA, T YOSHIKI, JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES AND HUMAN RETROVIROLOGY, 10, 2, 230, 230, Oct. 1995
LIPPINCOTT-RAVEN PUBL, English, Summary international conference

HTLV-I INDUCED MYELONEUROPATHY IN WKAH RATS - APOPTOSIS AND LOCAL ACTIVATION OF HTLV-I PX AND THE TNF-ALPHA GENES IMPLICATED IN THE PATHOGENESIS
U TOMARU, H IKEDA, O OHYA, T KASAI, YAMASITA, I, K MORITA, A WAKISAKA, T YOSHIKI, AIDS RESEARCH AND HUMAN RETROVIRUSES, 11, S127, S127, 1995
MARY ANN LIEBERT INC PUBL, English, Summary international conference

Inflammagingにおける炎症・老化細胞の解析とバイオマーカーへの展開
科学研究費助成事業
01 Apr. 2022 - 31 Mar. 2026
外丸 詩野, 石津 明洋
日本学術振興会, 基盤研究(B), 北海道大学, 22H02837

Reveal of the pathogenesis of MPO-ANCA-associated vasculitis and development of novel therapeutic strategies
Grants-in-Aid for Scientific Research
01 Apr. 2021 - 31 Mar. 2024
石津 明洋, 益田 紗季子, 外丸 詩野, 中沢 大悟
①MPO-ANCAはどのように好中球を活性化するのかについて、TNF-αでプライミングした好中球に液相または抗原固相により免疫複合体を形成した状態でMPO-ANCAを反応させ、好中球細胞外トラップ（NETs）の形成と細胞内シグナル分子のリン酸化を指標として好中球の活性化を評価した。その結果、どちらの場合もMPO-ANCAはNETsを誘導したが、抗原固相により免疫複合体を形成した状態で好中球に反応させた場合にシグナル分子がリン酸化されることを確認した。
②MPO-ANCAにより誘導されるNETsに血管炎惹起性はあるかについて、MPO-ANCA誘導モデルの病変糸球体にNETsの沈着があることを証明した。
③MPO-ANCA関連血管炎（MPO-AAV）におけるNETs除去障害の原因は何かについて、MPO-AAVの肺病変部に認められたDNase I抵抗性NETsと結核の肺病変部に認められたDNase I感受性NETs、そしてin vitroで誘導したDNase I感受性あるいは抵抗性NETsを用いてプロテオーム解析を行った。NETsにDNase I抵抗性をもたらす候補タンパク18種を抽出し、これらのリコンビナントタンパクをNETs誘導時に添加したところ、5つのタンパクで通常のNETsではなく類円形のDNase I抵抗性NETsが形成された。さらに、そのうちの一つのタンパクに対する阻害抗体をNETs誘導時に加えると、類円形NETsおよび通常型NETsの形成が抑制された。
これらの知見に基づき、MPO-AAVの病態形成機序に即した分子標的治療を開発するため、前臨床試験を開始した。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 21H02958

Analysis of inflammatory cells in age-related chronic inflammation
Grants-in-Aid for Scientific Research
30 Jul. 2020 - 31 Mar. 2022
Utano Tomaru
Accumulation of age-related abnormal T cells induces chronic inflammation, which is involved in the pathogenesis of various age-related diseases such as lifestyle-related diseases and cancer. In this study, we investigated that the alteration of self-antigens in aging induced chronic inflammation. Using oxidative stress-fragile mice showing age-related phenotypes, we found that altered self-antigens caused by cellular stress induced increased number of CD44highCD122highCD49dhigh CD8+ T cells and chronic inflammation.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Research (Exploratory), Hokkaido University, 20K21512

Molecular pathology of autoreactive inflammatory T cells
Grants-in-Aid for Scientific Research
01 Apr. 2018 - 31 Mar. 2022
Tomaru Utano
The immune system is an important biological defense system for protecting the body from pathogens, but abnormal autoreactive immune responses develop autoimmune diseases and inflammatory diseases. In this study, to develop a useful biomarker for autoreactive inflammatory cells and pathological evaluation of autoimmune disease, we established gene-modified mouse model expressing ubiquitous self-antigens (USAs-Tg). Using this model, we found that autoreactive CD8+ cells showed memory phenotype with PD-1+ Lag-3+ Tim-3+ CTLA-4-. In addition, the activation state of antigen-presenting cells affects the differentiation of inflammation-inducing cells.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 18H02629

Involvement of neutrophil extracellular traps in the development of diabetic angiopathy
Grants-in-Aid for Scientific Research
01 Apr. 2015 - 31 Mar. 2017
Akihiro Ishizu
Recent studies have demonstrated that high-dose glucose induces neutrophil extracellular trap (NET) formation which can be a risk for microvascular disorders. Thus, we attempted to determine the correlation of circulating NET levels and clinical/laboratory parameters in well-controlled T2D patients and to reveal the mechanism of NET formation induced by high-dose glucose. As a result, serum MPO-DNA complex levels that represent the NET formation in vivo were significantly higher in some well-controlled T2D patients in correlation with the clinical/laboratory parameters, which have been regarded as risk markers for microvascular complications. The aldose reductase inhibitor, ranirestat, could inhibit the NET formation induced by high-dose glucose. These findings suggest that elevated levels of circulating NETs can be a risk marker for microvascular complications in well-controlled T2D patients, and that the polyol pathway is involved in the NET formation induced by high-dose glucose.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 15K15105

Basic and clinical strategy to overcome microscopic polyangiitis
Grants-in-Aid for Scientific Research
01 Apr. 2014 - 31 Mar. 2017
Ishizu Akihiro, Arimura Yoshihiro, Sada Ken-ei
In vitro neutrophil extracellular trap (NET) formation was inhibited significantly by a peptidylarginine deiminase (PAD) inhibitor Cl-amidine. Serum MPO-ANCA titers of mice given Cl-amidine were significantly lower than those of control mice. The amounts of peritoneal NETs in the mice given Cl-amidine was significantly smaller than those in control mice.
Response to remission induction therapy can be predicted by monitoring the altered expressions of the 16 genes, including IRF7, IFIT1, IFIT5, OASL, CLC, GBP-1, PSMB9, HERC5, CCR1, CD36, MS4A4A, BIRC4BP, PLSCR1, DEFA1/DEFA3, DEFA4, and COL9A2, in the peripheral blood at an early point of treatment in MPA patients with high sensitivity (85.7%) and specificity (96.9%).
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 26293082

The role of proteasome in pulmonary fibrosis
Grants-in-Aid for Scientific Research
01 Apr. 2013 - 31 Mar. 2016
Nagai Katsura, TOMARU Utano
The activity of 20S proteasome in bronchoalveolar labage fluid tend to be higher in the patients with idiopathic pulmonary fibrosis (IPF) than normal subjects or sarcoidosis patients. Increased proteasome activity suggests a link to fibrotic reaction. In contrast, cigarette smoke-exposed Tg mice with decreased proteasomal chymotrypsin-like activity showed remarkable air space enlargement and increased apoptotic cells compared with wild type controls. Impaired proteasomal activity also enhanced apoptosis and endoplasmic reticulum stress in cigarette smoke extract (CSE)-exposed fibroblastic cells.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 25461181

Dysfunction of proteasome and pathogenesis in age-related disorders
Grants-in-Aid for Scientific Research
01 Apr. 2013 - 31 Mar. 2016
Tomaru Utano, ISHIZU AKIHIRO
The proteasome plays a critical role in proteostasis by rapidly destroying misfolded and modified proteins generated by cellular stresses. Proteasome activity decreases with aging in many organs, and an age-related decline in proteasomal function has been implicated in various age-related pathologies. Recently, we have established a transgenic (Tg) mouse model with decreased proteasomal chymotrypsin-like activity, showing age-related phenotypes. Using this model, we demonstrate here that decreased proteasomal function accelerates cigarette smoke (CS)-induced pulmonary emphysema. CS-exposed Tg mice showed remarkable airspace enlargement and increased foci of inflammation compared with wild-type controls. Collective evidence suggests that CS exposure and impaired proteasomal activity coordinately enhance apoptotic cell death in the alveolar walls that may be involved in the development and progression of emphysema in susceptible individuals such as the elderly.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 25460489

Analysis of retinal degeneration caused by dysfunction of ubiquitin proteasome system
Grants-in-Aid for Scientific Research
01 Apr. 2013 - 31 Mar. 2016
NODA Mika, NODA Kousuke, TOMARU Utano, OZAWA Yoko
Recent studies have revealed that ubiquitin proteasome system (UPS) dysfunction is involved in the pathogenesis of systemic neurodegenerative diseases such as Alzheimer’s disease. In ocular diseases, retinitis pigmentosa (RP) is a heterogeneous group of inherited retinal neurodegenerative disorders characterized by progressive degeneration of photoreceptors. Therefore, whether impairment of UPS function also causes RP has been of great interest.
In the present study, using an animal model of decreased proteasomal activity (β5t-Tg mice) we demonstrated that i) decreased function of proteasome causes retinal degeneration in vivo, largely attributed to photoreceptor cell death, and ii) photoreceptor cells are vulnerable to the functional disturbance of proteasome in the retina. The current data raise an interesting question regarding the impact of proteasome dysfunction in the pathogenesis of RP.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 25462702

Development of regenerative medicine for spinal cord injury in HTLV-1 associated myelopathy
Grants-in-Aid for Scientific Research
2010 - 2012
YAMANO Yoshihisa, TOMARU Utano
We studied somatic stem cells in order to forward the development of regenerative medicine for spinal cord injury in Human T-Lymphotropic Virus Type 1 (HTLV-1) associated myelopathy (HAM). We investigated the infectiousness of HTLV-1 to somatic stem cells and used an animal model for HAM to analyze the regenerative capacity of somatic stem cells as a treatment option. We transplanted adipose derived somatic stem cells (ASCs) from HTLV-1 infected GFP rats into GFP-negative non-infected rats, and no infection was observed. However, when we analyzed the infectiousness to human-derived ASCs in vitro, we clearly observed infection, indicating that there is a risk that ASCs could become infected with HTLV-1. We also attempted to create an animal model of early-onset HAM using HTLV-1 infected rats by challenging the infection every 2 weeks; however, no acceleratory effect was observed. Finally, we analyzed the major pathogenic processes leading to spinal cord lesions in HAM, and we demonstrated that overproduction of CXCL10 from astrocytes plays a major role in HAM pathogenesis.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), St. Marianna University School of Medicine, 22590943

Proteasomal dysfunction and pathology
Grants-in-Aid for Scientific Research
2010 - 2012
TOMARU Utano, ISHIZU Akihiro
In this study, we established a transgenic (β5t-Tg) mouse model with decreased proteasomal chymotrypsin-like activity. β5t-Tg mice exhibited a shortened lifespan and age-related phenotypes. When β5t-Tg mice were fed a high-fat diet, they developed more pronounced obesity and hepatic steatosis than wild-type mice. It has been known that proteasome plays an important role in the immune system, and β5t-Tg mice show abnormal T cell maturation. Our results demonstrate that decreased proteasomal chymotrypsin-like activity affects longevity and aggravates age-related metabolic disorders such as obesity and hepatic steatosis, and may correlate to the pathogenesis of immune-related disorders
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 22590353

Pathogenesis of vasculitis and novel diagnostic and therapeutic strategies
Grants-in-Aid for Scientific Research
2008 - 2010
ISHIZU Akihiro, TOMARU Utano, IWASAKI Sari
Autoreactive vasculitogenic T cell clones were established from env-pX rats, which develop medium to small-sized vasculitis. Identification of the T cell target and the mechanism of vascular injury induced by the T cells might result in the development of novel therapeutic strategies for vasculitis. The transcriptome analysis was performed using the peripheral blood obtained from patients with microscopic polyangiitis (MPA) before and 1 week after the treatment. Sixteen genes were nominated as distinguished indicators that could predict prognosis of patients with MPA at an early point during the therapy.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 20590390

Role of stress-inducible NKG2D ligands in disease
Grants-in-Aid for Scientific Research
2007 - 2008
KASAHARA Masanori, TOMARU Utano, FUGO Kazunori
NKG2Dリガンドはがん化やウイルス感染に伴って、細胞表面に発現される主要組織適合遺伝子複合体クラスI様分子であり、免疫系に異常細胞の存在を知らせる危険信号の働きをしている。本研究では、我々が同定した2個の新規リガンドを中心として、NKG2Dリガンドの構造、機能、病態における役割を解析した
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 19390107

慢性ウイルス感染症におけるメモリーT細胞の免疫応答と関連疾患の病態解明
科学研究費助成事業
2005 - 2006
外丸 詩野
本研究では、ヒトレトロウイルスであるヒトT細胞白血病ウイルス1型(HTLV-I)に着目し、HTLV-I関連疾患である脊髄症(HAM/TSP)を発症した患者におけるT細胞免疫応答を解析した。HAM/TSPではT細胞による過剰な免疫応答が疾患発症に関連している可能性が高く、また免疫抗原となるウイルスペプチドの研究が進んでおり、慢性ウイルス感染症におけるT細胞の免疫応答と関連疾患発症のメカニズムを研究するモデルとして有用である。本研究計画では、申請者が最近樹立した人工的抗原提示細胞を用いた解析法を応用し、ウイルス抗原に対する(1)CD8+T細胞による免疫応答、(2)CD4+T細胞による免疫応答、を患者血液から樹立したT細胞クローン、あるいは末梢血を用いてした。
研究期間では、(1)に関してウイルス抗原に対するCD8+T細胞による免疫応答の詳細な解析を行い、HAM/TSP患者では他の慢性ウイルス感染症に比べ、CD8+T細胞において細胞障害活性の低いintermediateな分化段階を示すメモリー細胞が集積している傾向にあることがわかった。また、(2)に関しては、新たに樹立したHLA-DRB1^*0101-GFP分子を発現する人工的抗原提示細胞のコントロール実験を行った。樹立したHLA-DRB1^*0101-GFP分子を発現する人工的抗原提示細胞では良好なHLA-DRB1^*0101-GFP分子の細胞表面での発現が観察され、蛍光顕微鏡による解析ではCD4+T細胞との接着1や細胞反応が認められた。現在、ウイルス抗原特異的CD4+T細胞の機能解析を継続的に行っている。今後は研究期間内での研究結果をふまえて、慢性ウイルス感染症におけるウイルス感染のコントロール機構や免疫破綻のメカニズムを解析し、新たな発症予防.治療戦略を展開する。
日本学術振興会, 若手研究(B), 北海道大学, 17790251

MILL分子群による免疫監視機構の解析
科学研究費助成事業
2005 - 2006
笠原 正典, 石津 明洋, 外丸 詩野
NKG2Dリガンドは癌細胞やウイルス感染細胞などで発現されるストレス誘導性のMHCクラスI様分子であり,NK細胞,CD8_+T細胞による免疫監視において重要な役割を果たしている.マウスでは,既知のリガンドとして,RAE-1α,-β,-γ,-δ,-ε,H60(Histocompatibility-60),MULT-1が知られている.本年度は,ヒトのNKG2Dリガンド分子の一つであるMICA-MICB分子と最も近縁なマウスのMHCクラスI様分子MILL(MHCclass I-like located near the leukocyte recptor complex)がNKG2Dのリガンドとして機能するか否かを検討するとともに,新しいマウスNKG2Dリガンドの同定を試みた.その結果,MILLは配列的にはNKG2Dのリガンドとして機能することが予想されたにもかかわらず,NKG2D-Ig融合タンパク質と結合しないことが示された.さらに,MILLはβ2ミクログロブリンと会合するGPIアンカー型タンパク質であり,この点でもMICA・MICB分子とは異なっていることが判明した.この結果を受けて新しいマウスNKG2Dリガンドを探索した結果,2個のMHCクラスI様分子がNKG2Dリガンドとして同定された.これらのNKG2Dリガンドは既知のリガンドの中ではH60と最も相似性が高かったため,H60b, H60cと命名した.H60b, H60c遺伝子の発現はサイトメガロウイルス感染や抗がん剤処理などのストレスによって顕著に誘導された.H60b, H60cのNKG2Dとの結合親和性はH60に較べて低く,H60cの結合親和性はH60bのそれより低かった.
日本学術振興会, 特定領域研究, 北海道大学, 17047001

EXPRESSION OF HUMAN ENDOGENOUS RETROVIRUS AND ITS POSSIBLE ROLES FOR DEVELOPING AUTOIMMUNE DESIASES
Grants-in-Aid for Scientific Research
2003 - 2004
TOMARU Utano, ISHIZU Akihiro, SUZUKI Akira
The aims of this project were to investigate the biological role of human endogenous retrovinis-R (HERV-R) in vivo. We have established transgenic rats carrying a full sequence of HERV-R under control of its own long terminal repeat promoter (HERV-R rat), and analyzed HERV-R expression and its possible roles for developing autoirnmune disease. Major research accomplishments were as follows :
1.The product of the transgene was detected as an 85 kDa glycoprotein in specific organs such as Harderian gland, prostatic gland, skin and salivary gland.
2.We have found that skin tissues transplanted from HERV-R rats to wild rats are rejected around 50 days after transplantation, suggesting that HERV-R antigen is recognized as a target antigen in the immune responses.
3.Because we observed no pathological significance in the HERV-R rats, we have established double transgenic rats carrying both human infectious retrovirus HTLV-I env-pX gene and HERV-R gene. Based on our previous studies, transgenic rat carrying HTLV-I env-pX gene indicate significant pathological findings which are similar to human autoimmune disorders. We have postulated that HERV-R antigen may modulate immune responses in the rats with a background of auto-immune responses. We observed that double transgenic rats had a tendency to be short-lived, and we found deposition of IgM-type autoantibodies in the Harderian gland. We also found thrombus formation in the glomeruli of kidney by histopathological analysis.
4.Based on our experiments, we propose a possibility that expression of endogenous retrovirus may modulate immune responses in the inflammatory diseases including autoimmune disorders.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), HOKKAIDO UNIVERSITY, 15590330

動物モデルを用いたHTLV-1発がん機構の解析
科学研究費助成事業
2003 - 2003
吉木 敬, 石津 明洋, 池田 仁, 志田 壽利, 外丸 詩野
lckプロモーターの制御下にヒトT細胞白血病ウイルスI型(HTLV-I)のpX遺伝子を発現するlck-pXラットは上皮型胸腺腫を発症する。このラットの腎被膜下に腫瘍化前のlck-pXラット胸腺を移植することにより、胸腺腫の悪性化と考えられる現象が観察された。この悪性胸腺腫では、オリジナルの胸腺腫で認められるp16とARFの発現が認められず、p16/ARF領域遺伝子の欠失が本腫瘍の悪性化に中心的な役割を果たしていると考えられた。HTLV-Iが原因となる成人T細胞白血病(ATL)は一般にキャリア状態から腫瘍化し、くすぶり型や慢性型といった低悪性度のATLから急性型やリンパ腫型といった高悪性度のATLへと進展する経過をたどる。高悪性度のATLへの移行の際にはp16やp53の遺伝子異常を高率に伴っていることが知られている。従って、lck-pXラットでの胸腺腫の悪性化はヒトATLの急性転化でみられる現象の一部を観察している可能性がある。良性腫瘍の状態から高悪性度腫瘍の状態までを連続的に観察できる本実験系はHTLV-I関連腫瘍の悪性化機構を解明する上で有用なin vivoモデルである。
一方、HTLV-Iがヒト以外の動物で効率よく増殖できないのは、ウイルス粒子の形成に必要なRexの働きを支持する宿主因子CRM1が、ヒト以外の動物ではHTLV-Iに対して低い支持活性しか示さないためであると考えられている。実際にHTLVV-1持続感染ラットT細胞株にヒトcrm1遺伝子を導入することにより、HTLV-I p19発現が増加した。ヒトcrm1遺伝子導入ラットはHTLV-1に対する高感染感受性宿主となる可能性が期待される。
日本学術振興会, 特定領域研究, 北海道大学, 15023202

Development and analyses of novel animal models for human retrovirus-induced desiases.
Grants-in-Aid for Scientific Research
2002 - 2003
KYOSHIKI Takashi, TOMARU Utano, ISHIZU Akihiro, IKEDA Hitoshi
Virus-host interactions were investigated using rat models for human retrovirus-induced diseases.
1.HAM rat disease occurred in HTLV-I-infected rats in a strain-dependent manner. In HAM susceptible rats, expression of the INF-γ gene in the spinal cord did not increase after HTLV-I infection, while did in HAM resistant strains. These findings suggest that INF-γ may play a role in prevention at HTLV-1-induced diseases.
2.Autoimmune diseases occurred in HTLV-I LTR-env-pX transgenic rats (LTR-env-pX rats). Functional alteration of CD25+CD4+ immunoregulatory T-cells was evident in LTR-env-pX rats before they developed diseases. Expressions of the CTLA-4, GITR, Foxp3, and SOCS family genes in the cells of LTR-env-pX rats were lower than those of wild type rats. Synovial tissues rather than bone marrow cells carrying the transgene may play roles in prolongation of arthritis developed in LTR-env-pX rats.
3.Benign epithelial thymoma occurred in lck-pX transgenic rats (lck-pX rats). Thymoma grafted in the subcapsular space of the kidney of lck-pX rats showed malignant transformation. The p16/ARF genes were lost in the transformed tumors, thus suggesting that these molecules may be associated with malignant transformation of HTLV-I-induced neoplasms.
4.Rat fibrobalastic cells expressing human CD4 and CXCR-4/CCR5 could be infected with HIV-1. Although HIV-1 provirus was integrated in the genome of these cells, viral expression was not achieved completely. Putative factors including human cyclin T1 and MHC class II transactivator (C2TA) may be needed for viral replication in the cells. To examine interactions of HIV-1 and host in vivo, transgenic rats carrying the human CD4, CXCR4/CCR5, cyclic T1, and C2TA genes were established.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), HOKKAIDO UNIVERSITY, 14207010