研究者データベース

外丸 詩野(トマル ウタノ)
医学研究院 病理系部門 病理学分野
准教授

基本情報

所属

  • 医学研究院 病理系部門 病理学分野

職名

  • 准教授

学位

  • 博士(医学)(北海道大学)

ホームページURL

J-Global ID

研究キーワード

  • 病理学   免疫学   実験病理学   

研究分野

  • ライフサイエンス / 実験病理学

職歴

  • 2012年11月 - 現在 北海道大学大学院 医学研究院 分子病理学教室 准教授
  • 2004年11月 - 2012年10月 北海道大学大学院 医学研究院 分子病理学教室 講師
  • 2003年04月 - 2004年10月 北海道大学大学院 医学研究院 分子病理学教室 助教

研究活動情報

論文

  • 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 2021年05月 
    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.
  • Tamihiro Kawakami, Yuto Tamura, Yupeng Dong, Miku Yoshinari, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
    The Journal of dermatology 48 5 703 - 706 2021年05月 
    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.
  • 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 2021年04月 
    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.
  • 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 2021年02月 
    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.
  • 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 2021年01月 
    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.
  • 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年 
    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.
  • 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 2020年11月 [査読有り][通常論文]
     
    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.
  • 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 2020年08月 
    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.
  • Ryo Uozumi, Risa Iguchi, Sakiko Masuda, Yuka Nishibata, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
    Modern rheumatology 30 3 544 - 550 2020年05月 [査読有り][通常論文]
     
    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.
  • 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 2020年01月27日 [査読有り][通常論文]
     
    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.
  • 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年 [査読有り][通常論文]
     
    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.
  • 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 2019年12月21日 [査読有り][通常論文]
  • 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 2019年08月 [査読有り][通常論文]
     
    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.
  • 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 2019年07月01日 [査読有り][通常論文]
     
    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.
  • 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 2019年04月 [査読有り][通常論文]
     
    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.
  • 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 2019年04月 [査読有り][通常論文]
     
    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.
  • Daigo Nakazawa, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
    Nature reviews. Rheumatology 15 2 123 - 123 2019年02月 
    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.
  • 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 2019年02月 [査読有り][通常論文]
     
    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.
  • Daigo Nakazawa, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
    Nature reviews. Rheumatology 15 2 91 - 101 2019年02月 [査読有り][通常論文]
     
    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.
  • Utano Tomaru, Saori Konno, Syota Miyajima, Rikuto Kimoto, Mari Onodera, Shizuka Kiuchi, Shigeo Murata, Akihiro Ishizu, Masanori Kasahara
    Cell reports 26 3 639 - 651 2019年01月15日 [査読有り][通常論文]
     
    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.
  • 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 2018年11月 [査読有り][通常論文]
     
    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.
  • Park JK, Otsuka N, Tomaru U, Suzuki H, Azuma M, Okamoto K, Yamashiro K, Kasahara M
    Human pathology 80 104 - 112 2018年10月 [査読有り][通常論文]
  • 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  2018年06月22日 [査読有り][通常論文]
     
    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.
  • 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 2018年04月01日 [査読有り][通常論文]
     
    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.
  • 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 2018年04月01日 [査読有り][通常論文]
     
    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.
  • Yusuke Nishioka, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
    Frontiers in Immunology 9 548  2018年03月15日 [査読有り][通常論文]
     
    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.
  • Emika Futamata, Sakiko Masuda, Yuka Nishibata, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu
    Nephron 138 4 328 - 330 2018年03月01日 [査読有り][通常論文]
  • 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 2017年08月 [査読有り][通常論文]
     
    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.
  • Mai Yamada, Tamihiro Kawakami, Kohei Takashima, Yusuke Nishioka, Yuka Nishibata, Sakiko Masuda, Shigeru Yoshida, Utano Tomaru, Akihiro Ishizu
    RHEUMATOLOGY 56 6 1013 - 1018 2017年06月 [査読有り][通常論文]
     
    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.
  • 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  2017年05月 [査読有り][通常論文]
     
    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.
  • 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 2017年02月 [査読有り][通常論文]
     
    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.
  • Yusuke Nishioka, Madoka Yamaguchi, Ai Kawakami, Maya Munehiro, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu
    AMERICAN JOURNAL OF PATHOLOGY 187 1 176 - 186 2017年01月 [査読有り][通常論文]
     
    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.
  • 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  2016年12月 [査読有り][通常論文]
     
    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.
  • Saldko Masuda, Daigo Nakazawa, Haruki Shida, Anna Miyoshi, Yoshihiro Kusunoki, Utano Tomaru, Akihiro Ishizu
    CLINICA CHIMICA ACTA 459 89 - 93 2016年08月 [査読有り][通常論文]
     
    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/).
  • 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 2016年06月 [査読有り][通常論文]
     
    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.
  • Daigo Nakazawa, Haruki Shida, Yoshihiro Kusunoki, Arina Miyoshi, Saori Nishio, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
    JOURNAL OF AUTOIMMUNITY 67 19 - 28 2016年02月 [査読有り][通常論文]
     
    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.
  • 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 2016年 [査読有り][通常論文]
  • 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 2016年 [査読有り][通常論文]
     
    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
  • 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 2015年11月 [査読有り][通常論文]
  • 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 2015年11月 [査読有り][通常論文]
  • Sari Iwasaki, Akira Suzuki, Takashi Fujisawa, Taiju Sato, Shinya Shirai, Mitsunori Kamigaki, Noriyuki Otsuka, Utano Tomaru, Akihiro Ishizu
    CARDIOVASCULAR PATHOLOGY 24 6 408 - 410 2015年11月 [査読有り][通常論文]
     
    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.
  • 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 2015年08月28日 [査読有り][通常論文]
  • Tomaru U, Tsuji T, Kiuchi S, Ishizu A, Suzuki A, Otsuka N, Ito T, Ikeda H, Fukasawa Y, Kasahara M
    Histopathology 67 2 235 - 244 2015年08月 [査読有り][通常論文]
     
    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.
  • 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 2015年07月 [査読有り][通常論文]
  • 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 2015年06月 [査読有り][通常論文]
     
    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.
  • Atsushi Wanifuchi, Keisuke Taguchi, Yoshinori Ikehata, Yuichiro Kurimura, Yoshiki Hiyama, Utano Tomaru
    Acta Urologica Japonica 61 6 249 - 252 2015年06月01日 [査読有り][通常論文]
     
    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.
  • 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 2015年02月 [査読有り][通常論文]
     
    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.
  • 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 2015年 [査読有り][通常論文]
     
    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
  • Matsui Y, Tomaru U, Miyoshi A, Ito T, Fukaya S, Miyoshi H, Atsumi T, Ishizu A
    Experimental and molecular pathology 97 3 354 - 358 2014年12月 [査読有り][通常論文]
     
    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
    2014年10月 [査読有り][通常論文]
  • 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 2014年08月 [査読有り][通常論文]
     
    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
  • 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 2014年08月 [査読有り][通常論文]
     
    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.
  • 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 2014年07月03日 [査読有り][通常論文]
     
    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.
  • 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 2014年05月 [査読有り][通常論文]
     
    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.
  • 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 2014年03月 [査読有り][通常論文]
     
    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.
  • Yamada Y, Tomaru U, Ishizu A, Kiuchi T, Kasahara M, Matsuno Y
    Journal of clinical pathology 67 3 276 - 278 2014年03月 [査読有り][通常論文]
     
    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.
  • 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 2014年01月 [査読有り][通常論文]
     
    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.
  • T. Imamoto, D. Nakazawa, H. Shida, A. Suzuki, N. Otsuka, U. Tomaru, A. Ishizu
    CLINICAL AND EXPERIMENTAL RHEUMATOLOGY 32 1 149 - 150 2014年01月 [査読有り][通常論文]
  • Ryoko Hamano, Tomohisa Baba, Soichiro Sasaki, Utano Tomaru, Akihiro Ishizu, Mitsuhiro Kawano, Masakazu Yamagishi, Naofumi Mukaida
    European Journal of Immunology 44 4 1005 - 1015 2014年 [査読有り][通常論文]
     
    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.
  • Nakazawa D, Nishio S, Tomaru U, Atsumi T, Ishizu A
    Nihon Jinzo Gakkai shi 56 2 117 - 123 2014年 [査読有り][通常論文]
  • 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 2013年12月 [査読有り][通常論文]
  • Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
    CLINICAL AND EXPERIMENTAL NEPHROLOGY 17 5 631 - 633 2013年10月 [査読有り][通常論文]
     
    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.
  • Utano Tomaru, Masanori Kasahara
    Archivum Immunologiae et Therapiae Experimentalis 61 5 357 - 365 2013年10月 [査読有り][通常論文]
     
    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.
  • Sora Takeuchi, Satoko Kimura, Yoshinao Soma, Masashi Waki, Madoka Yamaguchi, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu, Tamihiro Kawakami
    RHEUMATOLOGY 52 9 1592 - 1598 2013年09月 [査読有り][通常論文]
     
    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.
  • 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 2013年09月 [査読有り][通常論文]
     
    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.
  • Rie Michimata, Hidemichi Watari, Utano Tomaru, Noriaki Sakuragi, Akihiro Ishizu
    Pathobiology 80 5 259 - 264 2013年06月 [査読有り][通常論文]
     
    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.
  • 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 2013年06月 [査読有り][通常論文]
  • 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 2013年05月 [査読有り][通常論文]
     
    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.
  • 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  2013年05月 [査読有り][通常論文]
     
    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.
  • 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 2013年02月 [査読有り][通常論文]
  • Fukaya S, Matsui Y, Tomaru U, Kawakami A, Sogo S, Bohgaki T, Atsumi T, Koike T, Kasahara M, Ishizu A
    Laboratory investigation; a journal of technical methods and pathology 93 1 72 - 80 2013年01月 [査読有り][通常論文]
     
    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
  • Masuda S, Iwasaki S, Tomaru U, Baba T, Katsumata K, Ishizu A
    Clinical & developmental immunology 2013 345745  2013年 [査読有り][通常論文]
     
    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.
  • 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 2013年 [査読有り][通常論文]
     
    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.
  • Masuda S, Iwasaki S, Tomaru U, Sato J, Kawakami A, Ichijo K, Sogo S, Baba T, Katsumata K, Kasahara M, Ishizu A
    PloS one 7 12 e52918  2012年12月 [査読有り][通常論文]
     
    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.
  • 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 2012年11月 [査読有り][通常論文]
     
    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.
  • Tomaru U, Yamada Y, Ishizu A, Kuroda T, Matsuno Y, Kasahara M
    Journal of clinical pathology 65 9 858 - 859 2012年09月 [査読有り][通常論文]
  • 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  2012年09月 [査読有り][通常論文]
     
    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.
  • Baba T, Badr Mel S, Tomaru U, Ishizu A, Mukaida N
    PloS one 7 7 e41154  2012年07月 [査読有り][通常論文]
     
    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.
  • Tanaka Y, Manabe A, Nakadate H, Kondoh K, Nakamura K, Koh K, Tomaru U, Kikuchi A, Komiyama T
    Leukemia research 36 5 560 - 564 2012年05月 [査読有り][通常論文]
  • 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 2012年04月01日 [査読無し][通常論文]
  • 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 2012年03月 [査読有り][通常論文]
  • 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 2012年03月 [査読有り][通常論文]
  • 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 2012年03月 [査読有り][通常論文]
     
    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)
  • Sutoh Y, Kondo M, Ohta Y, Ota T, Tomaru U, Flajnik MF, Kasahara M
    Immunogenetics 64 1 49 - 58 2012年01月 [査読有り][通常論文]
     
    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.
  • 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年 [査読有り][通常論文]
  • Daigo Nakazawa, Utano Tomaru, Chiho Yamamoto, Satoshi Jodo, Akihiro Ishizu
    FRONTIERS IN IMMUNOLOGY 3 333  2012年 [査読有り][通常論文]
     
    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 2011年09月 [査読有り][通常論文]
  • Yamada Y, Tomaru U, Ishizu A, Kiuchi T, Marukawa K, Matsuno Y, Kasahara M
    The American journal of surgical pathology 35 9 1296 - 1304 2011年09月 [査読有り][通常論文]
     
    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 2011年03月 [査読有り][通常論文]
  • Iwasaki S, Masuda S, Baba T, Tomaru U, Katsumata K, Kasahara M, Ishizu A
    Cytometry. Part A : the journal of the International Society for Analytical Cytology 79A 1 46 - 56 2011年01月 [査読有り][通常論文]
     
    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
  • Hidemichi Watari, Rie Michimata, Motoaki Yasuda, Akihiro Ishizu, Utano Tomaru, Ying Xiong, Mohamed K. Hassan, Noriaki Sakuragi
    PATHOBIOLOGY 78 4 220 - 226 2011年 [査読有り][通常論文]
     
    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
  • 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 2010年11月 [査読有り][通常論文]
     
    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.
  • 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 2010年06月 [査読有り][通常論文]
     
    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.
  • Akihiro Ishizu, Asami Abe, Yukiko Miyatake, Tomohisa Baba, Chihiro Iinuma, Utano Tomaru, Takashi Yoshiki
    MODERN RHEUMATOLOGY 20 2 134 - 138 2010年04月 [査読有り][通常論文]
     
    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 2010年03月 [査読有り][通常論文]
  • 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 2009年09月 [査読有り][通常論文]
     
    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.
  • 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 2009年05月 [査読有り][通常論文]
     
    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)
  • 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 2008年03月 [査読有り][通常論文]
     
    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.
  • 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 2008年02月 [査読有り][通常論文]
     
    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.
  • 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 2007年12月 [査読有り][通常論文]
     
    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.
  • 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 2007年08月 [査読有り][通常論文]
     
    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.
  • 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 2006年09月 [査読有り][通常論文]
     
    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.
  • 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 2006年07月 [査読有り][通常論文]
     
    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.
  • T Baba, A Ishizu, S Iwasaki, A Suzuki, U Tomaru, H Ikeda, T Yoshiki, M Kasahara
    BLOOD 107 5 2004 - 2012 2006年03月 [査読有り][通常論文]
     
    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.
  • 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 2005年10月 [査読有り][通常論文]
     
    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.
  • H Hayase, A Ishizu, H Ikeda, Y Miyatake, T Baba, M Higuchi, A Abe, U Tomaru, T Yoshiki
    INTERNATIONAL IMMUNOLOGY 17 6 677 - 684 2005年06月 [査読有り][通常論文]
     
    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.
  • T Baba, A Ishizu, H Ikeda, Y Miyatake, T Tsuji, A Suzuki, U Tomaru, T Yoshiki
    EUROPEAN JOURNAL OF IMMUNOLOGY 35 6 1731 - 1740 2005年06月 [査読有り][通常論文]
     
    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.
  • Y Yamano, N Takenouchi, HC Li, U Tomaru, K Yao, CW Grant, DA Maric, S Jacobson
    JOURNAL OF CLINICAL INVESTIGATION 115 5 1361 - 1368 2005年05月 [査読有り][通常論文]
     
    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.
  • Utano Tomaru, Yoshihisa Yamano, Steven Jacobson
    Analyzing T Cell Responses: How to Analyze Cellular Immune Responses against Tumor Associated Antigens 261 - 274 2005年 [査読有り][通常論文]
     
    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.
  • Y Yamano, CJ Cohen, N Takenouchi, K Yao, U Tomaru, HC Li, Y Reiter, S Jacobson
    JOURNAL OF EXPERIMENTAL MEDICINE 199 10 1367 - 1377 2004年05月 [査読有り][通常論文]
     
    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.
  • U Tomaru, H Ikeda, XY Jiang, O Ohya, T Yoshiki
    JOURNAL OF NEUROVIROLOGY 9 5 530 - 538 2003年10月 [査読有り][通常論文]
     
    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.
  • U Tomaru, Y Yamano, M Nagai, D Maric, PTP Kaumaya, W Biddison, S Jacobson
    NATURE MEDICINE 9 4 469 - 475 2003年04月 [査読有り][通常論文]
     
    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.
  • CJ Cohen, O Sarig, Y Yamano, U Tomaru, S Jacobson, Y Reiter
    JOURNAL OF IMMUNOLOGY 170 8 4349 - 4361 2003年04月 [査読有り][通常論文]
     
    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.
  • Y Yamano, U Tomaru, S Jacobson
    TWO DECADES OF ADULT T-CELL LEUKEMIA AND HTLV-I RESEARCH 50 171 - 182 2003年 [査読有り][通常論文]
     
    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.
  • T Yano, T Kishimoto, U Tomaru, Y Kawarada, H Kato, T Yoshiki, H Ishikura
    PATHOLOGY 35 1 75 - 78 2003年01月 [査読有り][通常論文]
     
    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.
  • 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 2002年01月 [査読有り][通常論文]
     
    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.
  • 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 2001年07月 [査読有り][通常論文]
     
    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.
  • Makoto Saito, Syuhei Hige, Hiroshi Takeda, Utano Tomaru, Masahiko Shibata, Masahiro Asaka
    Journal of Gastroenterology 36 12 842 - 847 2001年 [査読有り][通常論文]
     
    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.
  • XY Jiang, H Ikeda, U Tomaru, K Morita, Y Tanaka, T Yoshiki
    JOURNAL OF NEUROIMMUNOLOGY 106 1-2 105 - 113 2000年07月 [査読有り][通常論文]
     
    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.
  • O Ohya, H Ikeda, U Tomaru, Yamashita, I, T Kasai, K Morita, A Wakisaka, T Yoshiki
    APMIS 108 6 459 - 466 2000年06月 [査読有り][通常論文]
     
    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.
  • U Tomaru, H Ishikura, SI Kon, M Kanda, H Harada, T Yoshiki
    JOURNAL OF UROLOGY 162 2 496 - 497 1999年08月 [査読有り][通常論文]
  • 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 1999年02月 [査読有り][通常論文]
     
    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.

その他活動・業績

  • 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 2020年03月 [査読無し][通常論文]
     
    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.
  • 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 2017年09月 [査読無し][通常論文]
  • 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 2017年09月 [査読無し][通常論文]
  • 中智昭, 外丸詩野, 松野吉宏 病理と臨床 35 151‐160 2017年04月11日 [査読無し][通常論文]
  • Yusuke Nishioka, Madoka Yamaguchi, Ai Kawakami, Maya Munehiro, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu RHEUMATOLOGY 56 30 -31 2017年03月 [査読無し][通常論文]
  • Yoshihiro Kusunoki, Daigo Nakazawa, Haruki Shida, Fumihiko Hattanda, Arina Miyoshi, Sakiko Masuda, Saori Nishio, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu RHEUMATOLOGY 56 112 -113 2017年03月 [査読無し][通常論文]
  • 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 2015年10月 [査読無し][通常論文]
     
    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.
  • 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 2015年05月 [査読無し][通常論文]
  • W. Kobayashi, T. Katsurada, U. Tomaru, N. Sakamoto, M. Kasahara GUT 63 A177 -A177 2014年06月 [査読無し][通常論文]
  • Nakazawa Daigo, Shida Haruki, Tomaru Utano, Yoshida Masaharu, Nishio Saori, Atsumi Tatsuya, Ishizu Akihiro NEPHROLOGY 19 149 -150 2014年05月 [査読無し][通常論文]
  • D. Nakazawa, U. Tomaru, S. Jodo, S. Nishio, T. Atsumi, A. Ishizu PRESSE MEDICALE 42 (4) 755 -755 2013年04月 [査読無し][通常論文]
  • A. Ishizu, U. Tomaru, T. Murai, T. Yamamoto, T. Atsumi, T. Yoshiki, H. Makino, S. Ozaki PRESSE MEDICALE 42 (4) 733 -733 2013年04月 [査読無し][通常論文]
  • 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 2013年03月 [査読無し][通常論文]
  • Tamihiro Kawakami, Sora Takeuchi, Satoko Kimura, Yoshinao Soma, Masashi Waki, Madoka Yamaguchi, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu JOURNAL OF DERMATOLOGY 39 49 -49 2012年06月 [査読無し][通常論文]
  • Daigo Nakazawa, Saori Nishio, Sekiya Shibasaki, Utano Tomaru, Ishizu Akihiro NEPHROLOGY DIALYSIS TRANSPLANTATION 27 442 -442 2012年05月 [査読無し][通常論文]
  • Y. Yamada, U. Tomaru, K. Kubota, A. Ishizu, T. Kiuchi, T. Mitsuhashi, Y. Matsuno, M. Kasahara LABORATORY INVESTIGATION 92 493A -493A 2012年02月 [査読無し][通常論文]
  • Y. Yamada, U. Tomaru, K. Kubota, A. Ishizu, T. Kiuchi, T. Mitsuhashi, Y. Matsuno, M. Kasahara MODERN PATHOLOGY 25 493A -493A 2012年02月 [査読無し][通常論文]
  • Akihiro Ishizu, Chihiro Iinuma, Masashi Waki, Madoka Yamaguchi, Naomi Sasaki, Utano Tomaru, Takashi Yoshiki CLINICAL AND EXPERIMENTAL IMMUNOLOGY 164 128 -128 2011年05月 [査読無し][通常論文]
  • 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 2010年03月 [査読無し][通常論文]
  • 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 2009年06月 [査読無し][通常論文]
  • 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 2009年03月 [査読無し][通常論文]
  • 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 2009年01月 [査読無し][通常論文]
  • 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 2009年01月 [査読無し][通常論文]
  • 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 2008年10月 [査読無し][通常論文]
  • 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 2008年09月 [査読無し][通常論文]
  • 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 2008年09月 [査読無し][通常論文]
  • 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 2008年03月 [査読無し][通常論文]
  • 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 2008年03月 [査読無し][通常論文]
  • 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 2007年11月 [査読無し][通常論文]
  • 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 2007年08月 [査読無し][通常論文]
  • 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 2007年02月 [査読無し][通常論文]
  • 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 2007年02月 [査読無し][通常論文]
  • 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 2007年02月 [査読無し][通常論文]
  • 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 2007年02月 [査読無し][通常論文]
  • 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 2006年11月 [査読無し][通常論文]
  • 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 2006年04月 [査読無し][通常論文]
  • 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 2005年09月 [査読無し][通常論文]
  • 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 2004年05月 [査読無し][通常論文]
  • Y Yamano, CJ Cohen, U Tomaru, Y Reiter, S Jacobson AIDS RESEARCH AND HUMAN RETROVIRUSES 19 S38 -S38 2003年 [査読無し][通常論文]
  • KS Jones, U Tomaru, Y Yamano, C Sadowski, M Dambach, S Jacobson, FW Ruscetti AIDS RESEARCH AND HUMAN RETROVIRUSES 19 S28 -S28 2003年 [査読無し][通常論文]
  • U Tomaru, Y Yamano, M Nagai, S Jacobson AIDS RESEARCH AND HUMAN RETROVIRUSES 19 S8 -S9 2003年 [査読無し][通常論文]
  • U Tomaru, Y Yamano, M Nagai, D Maric, PT Kaumaya, W Biddison, S Jacobson AIDS RESEARCH AND HUMAN RETROVIRUSES 19 S44 -S45 2003年 [査読無し][通常論文]
  • Y Yamano, U Tomaru, M Nagai, S Jacobson AIDS RESEARCH AND HUMAN RETROVIRUSES 19 S41 -S41 2003年 [査読無し][通常論文]
  • 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 2002年01月01日 [査読無し][通常論文]
  • 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 1996年08月 [査読無し][通常論文]
     
    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.
  • 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 1995年10月 [査読無し][通常論文]
  • 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年 [査読無し][通常論文]

教育活動情報

主要な担当授業

  • 臨床病理学
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
  • 基本医学研究
    開講年度 : 2020年
    課程区分 : 修士課程
    開講学部 : 医学院
    キーワード : 病理学の基礎 Basic Pathology
  • 医学総論
    開講年度 : 2020年
    課程区分 : 博士後期課程
    開講学部 : 医学研究科
  • 基本医学総論
    開講年度 : 2020年
    課程区分 : 修士課程
    開講学部 : 医学院
    キーワード : 細胞と組織の基本的病的変化,疾患の種類と分類,疾患の病因と病態 Basic pathological changes in cells and tissues, types and classification of diseases, and causes and pathophysiology of diseases
  • 医学総論
    開講年度 : 2020年
    課程区分 : 博士後期課程
    開講学部 : 医学院
    キーワード : 疾患の分子機構、免疫、診断病理
  • 基盤医学研究
    開講年度 : 2020年
    課程区分 : 博士後期課程
    開講学部 : 医学院
    キーワード : 実験病理学、人体病理学
  • 保健組織病理学
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 循環器、呼吸器、消化器、造血器、内分泌器、泌尿・生殖器、運動器、皮膚・感覚器、神経系
  • 器官病理学
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 循環器、呼吸器、消化器、造血器、内分泌器、泌尿・生殖器、運動器、皮膚・感覚器、神経系
  • 細胞診断学講義演習
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 細胞診、細胞検査士(サイトスクリーナー)
  • 病理学
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 細胞と組織の基本的病的変化,疾患の種類と分類,疾患の病因と病態
  • 臨床病理学
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 臨床診断、病理診断、診断の根拠と鑑別診断、治療の評価
  • 病理学演習
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 肉眼的病理診断、組織学的病理診断、最終診断、個体の総合的病態、ポスター/スライド発表
  • 病理学実習
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 組織病理、光学顕微鏡、バーチャルスライド、免疫染色、スケッチ


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