Researcher Profile and Settings

Affiliation

• Faculty of Health Sciences　Health Sciences　Medical Laboratory Science

Job Title

• Lecturer

Research funding number

• 10763617

J-Global ID

• 201501050448302086

Research Areas

• Life sciences / Experimental pathology

Educational Organization

•  Bachelor's degree program　Departments of Health Sciences, School of Medicine
•  Master's degree program　Graduate School of Health Sciences
•  Doctoral (PhD) degree program　Graduate School of Health Sciences

Association Memberships

• JAPANESE ASSOCIATION OF MEDICAL TECHNOLOGISTS   JAPANESE SOCIETY OF LABORATORY MEDICINE   JAPAN COLLEGE OF RHEUMATOLOGY   THE JAPANESE SOCIETY OF CLINICAL CYTOLOGY   日本病理学会   

Research Activities

Published Papers

• Methylprednisolone pulse-enhanced neutrophil extracellular trap formation in mice with imiquimod-induced lupus-like disease, resulting in ischaemia of the femoral head cartilage.

  Hodaka Ogawa, Shunichi Yokota, Yumeka Hosoi, Ayano Shindo, Naho Ogawa, Ryodai Yamamura, Tomohiro Shimizu, Issei Nakade, Suishin Arai, Mai Taniguchi, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Norimasa Iwasaki, Akihiro Ishizu

  Lupus science & medicine 10 (2) 2023/12/28 

  OBJECTIVES: Methylprednisolone (mPSL) pulse therapy is an essential option for patients with active systemic lupus erythematosus, but there is a risk of adverse events related to microcirculation disorders, including idiopathic osteonecrosis of the femoral head (ONFH). Recent studies have revealed that excessive neutrophil extracellular traps (NETs) are involved in microcirculation disorders. This study aimed to demonstrate that mPSL pulse could induce NETs in lupus mice and identify the factors contributing to this induction. METHODS: Six mice with imiquimod (IMQ)-induced lupus-like disease and six normal mice were intraperitoneally injected with mPSL on days 39 to 41, and five mice with IMQ-induced lupus-like disease and six normal mice were injected with phosphate-buffered saline. Pathological examinations were conducted to evaluate the ischaemic state of the femoral head and tissue infiltration of NET-forming neutrophils. Proteome analysis was performed to extract plasma proteins specifically elevated in mPSL-administered mice with IMQ-induced lupus-like disease, and their effects on NET formation were assessed in vitro. RESULTS: Mice with IMQ-induced lupus-like disease that received mPSL pulse demonstrated ischaemia of the femoral head cartilage with tissue infiltration of NET-forming neutrophils. Proteome analysis suggested that prenylcysteine oxidase 1 (PCYOX1) played a role in this phenomenon. The reaction of PCYOX1-containing very low-density lipoproteins (VLDL) with its substrate farnesylcysteine (FC) induced NETs in vitro. The combined addition of IMQ and mPSL synergistically enhanced VLDL-plus-FC-induced NET formation. CONCLUSION: PCYOX1 and related factors are worthy of attention to understand the underlying mechanisms and create novel therapeutic strategies for mPSL-mediated microcirculation disorders, including ONFH.
• Bruton's tyrosine kinase is a possible therapeutic target in microscopic polyangiitis.

  Issei Nakade, Yuto Tamura, Fuyu Hashimoto, Yuko Ariza, Shingo Hotta, Hirofumi Fujigaya, Suishin Arai, Mai Taniguchi, Hodaka Ogawa, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu

  Arthritis research & therapy 25 (1) 215 - 215 2023/11/06 

  BACKGROUND: Bruton's tyrosine kinase (Btk) is an enzyme expressed in leukocytes other than T lymphocytes and plasma cells and involved in B-cell receptor- and Fcγ receptor (FcγR)-mediated signal transduction. Btk inhibitors potentially suppress autoantibody production due to the expected inhibitory ability of B lymphocyte differentiation into antibody-producing plasma cells and reduce FcγR-mediated neutrophil activation, including the release of neutrophil extracellular traps (NETs). Microscopic polyangiitis (MPA) is a systemic small-vessel vasculitis characterized by the pathogenic autoantibody, antineutrophil cytoplasmic antibody (ANCA) that reacts with myeloperoxidase (MPO). MPO and MPO-ANCA immune complex (IC)-induced FcγR-mediated NETs are critically involved in MPA pathogenesis. This study aimed to demonstrate the therapeutic efficacy of the Btk inhibitor tirabrutinib on MPA. METHODS: Various doses of tirabrutinib or vehicle were orally administered to Sprague-Dawley rats daily. Four weeks later, the number of peripheral B lymphocytes was counted, and Btk phosphorylation in B lymphocytes was evaluated by flow cytometry. Human peripheral blood neutrophils were stimulated by MPO and anti-MPO antibody ICs (MPO and anti-MPO-ICs), and Btk and its downstream Vav phosphorylation were assessed by western blotting. The effects of tirabrutinib on MPO and anti-MPO-IC-induced NET formation were examined in vitro. Wistar Kyoto rats were immunized with human MPO to induce experimental MPA and given drug-free or tirabrutinib-containing feed (0.0037% or 0.012%) from day 0 or 28. All rats were euthanized on day 42 for serological and histological evaluation. RESULTS: Tirabrutinib inhibited Btk phosphorylation without decreasing B lymphocytes in vivo. Neutrophil Btk and Vav were phosphorylated when stimulated with MPO and anti-MPO-ICs. Tirabrutinib suppressed MPO and anti-MPO-IC-induced NET formation in vitro and ameliorated experimental MPA in a dose-dependent manner in vivo. Although MPO-ANCA production was not affected, NET-forming neutrophils in the blood were significantly reduced by tirabrutinib. CONCLUSIONS: The Btk inhibitor tirabrutinib suppressed MPO and anti-MPO-IC-induced NET formation in vitro and ameliorated experimental MPA by reducing NET-forming neutrophils but not decreasing MPO-ANCA titer in vivo. This study suggests that Btk is a possible therapeutic target in MPA.
• Demonstration of equivocal anti-glomerular basement membrane antibody positivity as a non-specific reaction through multiple immunologic assays in a case of pediatric asymptomatic hematuria

  Masayuki Sato, Yuka Nishibata, Sakiko Masuda, Tsunehisa Nagamori, Emi Ishibazawa, Yoichiro Yoshida, Hironori Takahashi, Akihiro Ishizu, Satoru Takahashi

  Clinical Biochemistry 120 110650 - 110650 0009-9120 2023/10
• CD47 blockade ameliorates autoimmune vasculitis via efferocytosis of neutrophil extracellular traps.

  Satoka Shiratori-Aso, Daigo Nakazawa, Takashi Kudo, Masatoshi Kanda, Yusho Ueda, Kanako Watanabe-Kusunoki, Saori Nishio, Sari Iwasaki, Takahiro Tsuji, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu, Tatsuya Atsumi

  JCI insight 2023/06/27 

  Neutrophil extracellular trap (NET) formation contributes to immune defense and is a distinct form of cell death. Excessive NET formation is found in patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), contributing to disease progression. The clearance of dead cells by macrophages, a process known as efferocytosis, is regulated by the CD47-mediated "don't eat me" signal. Hence, we hypothesized that pathogenic NETs in AAV escape from efferocytosis via the CD47 signaling pathway, resulting in the development of necrotizing vasculitis. Immunostaining for CD47 in human renal tissues revealed high CD47 expression in crescentic glomerular lesions of patients with AAV. In ex vivo studies, ANCA-induced netting neutrophils increased the expression of CD47 with the reduction of efferocytosis. After efferocytosis, macrophages displayed pro-inflammatory phenotypes. The blockade of CD47 in spontaneous crescentic glomerulonephritis-forming/Kinjoh (SCG/Kj) mice ameliorated renal disease and reduced myeloperoxidase (MPO)-ANCA titers with a reduction in NETs formation. Thus, CD47 blockade would protect against developing glomerulonephritis in AAV via restored efferocytosis of ANCA-induced NETs.
• 無症候性血尿に対する精査で抗糸球体基底膜抗体が陽性だったが、抗体解析により非特異的反応と考えられた男児例

  佐藤 雅之, 西端 友香, 益田 紗季子, 長森 恒久, 石羽澤 映美, 吉田 陽一郎, 高橋 弘典, 石津 明洋, 高橋 悟

  日本小児腎臓病学会雑誌 (一社)日本小児腎臓病学会 36 (Suppl.) 192 - 192 0915-2245 2023/05
• ANCA関連血管炎に対するアバコパンを含む新規治療 C5a受容体拮抗薬と好中球エラスターゼ阻害剤の好中球活性化抑制比較

  荒井 粋心, 西端 友香, 谷口 舞, 益田 紗季子, 外丸 詩野, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 67回 629 - 629 2023/03
• ANCA関連血管炎に対するアバコパンを含む新規治療 Cathepsin C阻害による好中球細胞外トラップ形成抑制

  西端 友香, 荒井 粋心, 谷口 舞, 益田 紗季子, 外丸 詩野, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 67回 629 - 629 2023/03
• ANCA関連血管炎に対するアバコパンを含む新規治療 好中球細胞外トラップにDNase I抵抗性を付与するタンパクの同定

  谷口 舞, 益田 紗季子, 中村 哲朗, 荒井 粋心, 西端 友香, 外丸 詩野, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 67回 630 - 630 2023/03
• ベーチェット病 ベーチェット病における口内炎の発生原因の解明

  益田 紗季子, 西端 友香, 川邊 智宏, 宮前 多佳子, 外丸 詩野, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 67回 649 - 649 2023/03
• 自己抗体から紐解く疾患の病理病態 ANCA関連血管炎におけるintermolecular epitope spreadingによる抗GBM抗体の産生

  西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  日本病理学会会誌 (一社)日本病理学会 112 (1) 196 - 196 0300-9181 2023/03
• SLEモデルマウスへのステロイドパルスは好中球細胞外トラップを誘導する

  小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 67回 886 - 886 2023/03
• COVID-19関連を含むIgA血管炎皮膚生検標本を使用したNeutrophil Extracellular Traps(NETs)の検証

  川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 石津 明洋, 外丸 詩野

  日本皮膚科学会雑誌 (公社)日本皮膚科学会 133 (2) 254 - 254 0021-499X 2023/02
• 感染症と血管炎 COVID-19発症後およびCOVID-19ワクチン接種後IgA血管炎の皮膚生検組織における好中球細胞外トラップの沈着 COVID-19非関連IgA血管炎との比較

  益田 紗季子, 西端 友香, 外丸 詩野, 横山 華英, 池田 高治, 川上 民裕, 石津 明洋

  脈管学 (一社)日本脈管学会 63 (1) 10 - 11 0387-1126 2023/02
• 無症候性血尿を呈した抗糸球体基底膜(GBM)抗体陽性症例の血清を用いた抗体解析

  西端 友香, 佐藤 雅之, 長森 恒久, 益田 紗季子, 外丸 詩野, 石津 明洋

  脈管学 (一社)日本脈管学会 63 (1) 13 - 14 0387-1126 2023/02
• 皮膚血管炎動物モデルの完成

  川上 民裕, 中出 一生, 田村 宥人, 伊藤 吹夕, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  脈管学 (一社)日本脈管学会 63 (1) 15 - 15 0387-1126 2023/02
• Low disease activity of microscopic polyangiitis in patients with anti-myosin light chain 6 antibody that disrupts actin rearrangement necessary for neutrophil extracellular trap formation.

  Miku Yoshinari, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Yoshihiro Arimura, Koichi Amano, Yukio Yuzawa, Ken-Ei Sada, Tatsuya Atsumi, Hiroaki Dobashi, Hitoshi Hasegawa, Masayoshi Harigai, Seiichi Matsuo, Hirofumi Makino, Akihiro Ishizu

  Arthritis research & therapy 24 (1) 274 - 274 2022/12/16 

  BACKGROUND: Neutrophil extracellular traps (NETs) are critically involved in microscopic polyangiitis (MPA) pathogenesis, and some patients with MPA possess anti-NET antibody (ANETA). Anti-myosin light chain 6 (MYL6) antibody is an ANETA that affects NETs. This study aimed to determine the significance of anti-MYL6 antibody in MPA. METHODS: The influence of anti-MYL6 antibody on NET formation and actin rearrangement necessary for NET formation was assessed by fluorescent staining. An enzyme-linked immunosorbent assay was established to detect serum anti-MYL6 antibody, and the prevalence of this antibody in MPA was determined. Furthermore, the disease activity and response to remission-induction therapy of MPA were compared between anti-MYL6 antibody-positive and anti-MYL6 antibody-negative MPA patients. RESULTS: Anti-MYL6 antibody disrupted G-actin polymerization into F-actin, suppressing phorbol 12-myristate 13-acetate-induced NET formation. Serum anti-MYL6 antibody was detected in 7 of 59 patients with MPA. The Birmingham vasculitis activity score (BVAS) of anti-MYL6 antibody-positive MPA patients was significantly lower than anti-MYL6 antibody-negative MPA patients. Among the nine BVAS evaluation items, the cutaneous, cardiovascular, and nervous system scores of anti-MYL6 antibody-positive MPA patients were significantly lower than anti-MYL6 antibody-negative MPA patients, although other items, including the renal and chest scores, were equivalent between the two groups. The proportion of patients with remission 6 months after initiation of remission-induction therapy in anti-MYL6 antibody-positive MPA patients was significantly higher than in anti-MYL6 antibody-negative MPA patients. CONCLUSIONS: Collective findings suggested that anti-MYL6 antibody disrupted actin rearrangement necessary for NET formation and could reduce the disease activity of MPA.
• Similar deposition of neutrophil extracellular traps in the dermis among COVID-19-associated IgA vasculitis, post-COVID-19 vaccination IgA vasculitis, and COVID-19-unrelated IgA vasculitis.

  Tamihiro Kawakami, Kae Yokoyama, Takaharu Ikeda, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu

  The Journal of dermatology 50 (5) e151-e152  2022/12/15
• 正常ラットにヒストン皮下注射後,抗ホスファチジルセリン・プロトロンビン複合体抗体と抗リソソーム膜タンパク質2抗体の静脈注射により,皮膚血管炎の発症に成功した

  川上 民裕, 中出 一生, 田村 宥人, 伊藤 吹夕, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  日本皮膚免疫アレルギー学会総会学術大会プログラム・抄録集 (一社)日本皮膚免疫アレルギー学会 52回 215 - 215 2022/12
• A Novel Antineutrophil Extracellular Trap Antibody Targeting Myosin Light Chain 6 in Microscopic Polyangiitis.

  Miku Yoshinari, Fumihiko Hattanda, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu

  The Journal of rheumatology 49 (11) 1286 - 1288 2022/11
• ベーチェット病皮膚生検標本を使用したNETsの検証

  川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  西日本皮膚科 日本皮膚科学会-西部支部 84 (4) 371 - 371 0386-9784 2022/08
• ベーチェット病皮下の血栓性静脈炎におけるNeutrophil Extracellular Trapsの発現

  川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  アレルギー (一社)日本アレルギー学会 71 (6-7) 870 - 870 0021-4884 2022/08
• Cyclophilin D regulates NETosis and inflammation in myeloperoxidase-antineutrophil cytoplasmic antibody-associated vasculitis.

  Takashi Kudo, Daigo Nakazawa, Kanako Watanabe-Kusunoki, Masatoshi Kanda, Satoka Shiratori-Aso, Nobuya Abe, Saori Nishio, Jun-Ichiro Koga, Sari Iwasaki, Takahiro Tsuji, Yuichiro Fukasawa, Miwako Yamasaki, Masahiko Watanabe, Sakiko Masuda, Utano Tomaru, Masaaki Murakami, Yasuaki Aratani, Akihiro Ishizu, Tatsuya Atsumi

  Arthritis & rheumatology (Hoboken, N.J.) 75 (1) 71 - 83 2022/07/29 

  OBJECTIVE: Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is pathologically characterized by focal fibrinoid necrosis where ANCA-mediated neutrophil extracellular trap (NET) formation and subsequent endothelial necrosis occurs. Cyclophilin D (CypD) plays an important role in mediating cell necrosis and inflammation via opening of mitochondrial permeability transition pores (mPTP). Here, we examined the role of CypD in AAV pathogenesis. METHODS: In vitro, the role and mechanism of CypD in ANCA-stimulated neutrophils were assessed by immunostaining and electron microscopy. A comprehensive RNA sequencing analysis was performed on ANCA-treated murine neutrophils. To investigate the role of CypD in vivo, an-anti-MPO IgG-transfer AAV model or spontaneous AAV model mice were induced in CypD knockout or wild-type mice. RESULTS: In vitro, pharmacological and genetic inhibition of CypD suppressed ANCA-induced NET formation via the suppression of reactive oxygen species/cytochrome c release from the mitochondria. The analysis of RNA sequencing in ANCA-treated murine neutrophils revealed the involvement of inflammatory responses, and CypD deficiency reduced ANCA-induced alterations in gene expression. Furthermore, the upstream regulator analysis revealed the relevance of intracellular calcium (CypD activator) and cyclosporin (CypD inhibitor) in ANCA stimulation, indicating that CypD-dependent mPTP opening is associated with ANCA-induced neutrophil activation and NETosis. In both AAV models, the genetic deletion of CypD ameliorated crescentic glomerulonephritis via the inhibition of CypD-dependent neutrophil and endothelial necrosis. CONCLUSIONS: CypD targeting is a novel and specific therapeutic strategy for AAV via the resolution of necrotizing vasculitis.
• Typical cutaneous small-vessel vasculitis induced by combined injection of antiphosphatidylserine/prothrombin complex antibody and anti-LAMP-2 antibody in normal rats.

  Tamihiro Kawakami, Issei Nakade, Yuto Tamura, Fuyu Ito, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu

  The Journal of dermatology 49 (12) 1233 - 1237 2022/07/25 

  We previously reported that IgA vasculitis and cutaneous arteritis could be dependently associated with the presence of the antiphosphatidylserine/prothrombin complex (anti-PS/PT) antibody and lysosomal-associated membrane protein-2 (LAMP-2). The copy number of LAMP-2 mRNA in skin tissue samples from rats with cutaneous vasculitis induced by intravenous administration of anti-PS/PT antibody after subcutaneous histone injection was significantly higher than in those without cutaneous vasculitis. We found LAMP-2 protein overexpression in neutrophils and vascular endothelial cells of the affected blood vessels in rats with cutaneous vasculitis induced by intravenous administration of anti-PS/PT antibody after cutaneous priming by histones. We found typical cutaneous small-vessel vasculitis in the skin of rats given intravenous injection of both anti-PS/PT antibody and anti-LAMP-2 antibody after cutaneous priming by histones. We suggested that the introduction of skin local histones and anti-PS/PT antibody in serum could move LAMP-2 to the cell surface of neutrophils and vascular endothelial cells, and that anti-LAMP-2 antibody could bridge these cells through antigen-specific binding in typical cutaneous small-vessel vasculitis.
• Presence of neutrophil extracellular traps in superficial venous thrombosis of Behçet's disease.

  Tamihiro Kawakami, Kae Yokoyama, Takaharu Ikeda, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu

  The Journal of dermatology 49 (7) 741 - 745 2022/04/17 

  Behçet's disease (BD) has a heterogeneous spectrum of disease manifestations featuring the involvement of different organs and can be characterized with different symptoms depending on the clinical department in charge. We retrospectively reviewed BD patients seen at our hospital and investigated the presence of neutrophils producing neutrophil extracellular traps (NET) in those patients. Immunolabeling of myeloperoxidase and histone citrullination proteins was performed on skin biopsies from three BD patients who had skin biopsy-proven superficial vein thrombophlebitis in their erythema nodosum-like lesions. We observed a higher proportion of female patients and a higher incidence of acne-like eruptions among BD patients seen at our dermatology department, while there was a higher incidence of ocular and gastrointestinal involvement among BD patients treated in other departments. We suggest that sex statistical trends could lead to the co-development of different manifestations and may help clinicians choose the best therapeutic approaches, tailoring them to the specific phenotype of the patient rather than one based on single disease manifestations. NET were found in neutrophils of panniculitis concurrent with superficial vein thrombophlebitis. We suggest that the pathogenesis of BD-related thrombosis could be associated with neutrophil activation and NET are released in the panniculitis of affected skin lesions, erythema nodosum-like lesions.
• Phorbol 12-myristate 13-acetate stimulation under hypoxia induces nuclear swelling with DNA outflow but not extracellular trap formation of neutrophils.

  Sakiko Masuda, Kurumi Kato, Misato Ishibashi, Yuka Nishibata, Ayako Sugimoto, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Ichizo Tsujino, Akihiro Ishizu

  Experimental and molecular pathology 125 104754 - 104754 2022/03/05 

  Neutrophils stand sentinel over infection and possess diverse antimicrobial weapons, including neutrophil extracellular traps (NETs). NETs are composed of web-like extracellular DNA decorated with antimicrobial substances and can trap and eliminate invading microorganisms. Although phorbol 12-myristate 13-acetate (PMA) is a potent NET inducer, previous studies have demonstrated that not all neutrophils exhibit NET formation even if stimulated by PMA at high concentrations. This study first showed that some neutrophils stimulated by PMA displayed a swollen nucleus but not NET formation and that hypoxic environments suppressed the NET release. Next, characterization of PMA-stimulated neutrophils with a swollen nucleus was accomplished by differentiating between suicidal-type NETosis and apoptosis. Furthermore, the significance of the phenomenon was examined using formalin-fixed, paraffin-embedded human lung disease tissues with and without pneumonia. As a result, histone H3 citrullination, DNA outflow, propidium iodide labeling, resistance to DNase I, and suspended actin rearrangement were characteristics of PMA-stimulated neutrophils with a swollen nucleus distinct from neutrophils that underwent either suicidal-type NETosis or apoptosis. Neutrophils stimulated by PMA under hypoxic conditions secreted matrix metalloproteinase-9 cytotoxic to human lung-derived fibroblasts. Further, deposition of neutrophil-derived citrullinated histone H3+ chromatin substances in pulmonary lesions was greater in patients with pneumonia than in patients without pneumonia and positively correlated with hypoxia-inducible factor-1α expression. The collective findings suggested that neutrophils activated under hypoxic conditions could be putative modulators of hypoxia-related disease manifestations.
• 変貌する糸球体疾患の概念:近年のトピックス 膜性腎症の特異抗原(PLA2R,THSD7A,NELL1,EXT1/2)について最近の話題

  辻 隆裕, 牧田 啓史, 深澤 雄一郎, 加賀 幸斗, 西端 友香, 益田 紗季子, 石津 明洋, 岩崎 沙理

  日本病理学会会誌 (一社)日本病理学会 111 (1) 162 - 162 0300-9181 2022/03
• 好中球細胞外トラップにDNase I抵抗性を付与するタンパクの探索

  益田 紗季子, 北野 翔大, 西端 友香, 外丸 詩野, 石津 明洋

  日本病理学会会誌 (一社)日本病理学会 111 (1) 214 - 214 0300-9181 2022/03
• SLE・抗リン脂質抗体症候群:基礎 全身性エリテマトーデスへのステロイドパルスが好中球細胞外トラップ形成に及ぼす影響

  小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 66回 347 - 347 2022/03
• ANCA関連血管炎:基礎研究・予後予測因子 ブルトン型チロシンキナーゼ阻害剤チラブルチニブによるMPO-ANCA関連血管炎誘導モデルの発症抑制

  中出 一生, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 66回 406 - 406 2022/03
• ANCA関連血管炎:基礎研究・予後予測因子 MPO-ANCA関連血管炎モデルにおける新規好中球機能制御化合物薬の抑制効果

  西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 66回 407 - 407 2022/03
• SLE・抗リン脂質抗体症候群:基礎 全身性エリテマトーデスへのステロイドパルスが好中球細胞外トラップ形成に及ぼす影響

  小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋

  日本リウマチ学会総会・学術集会プログラム・抄録集 (一社)日本リウマチ学会 66回 347 - 347 2022/03
• Possible implication of intermolecular epitope spreading in the production of anti-glomerular basement membrane antibody in anti-neutrophil cytoplasmic antibody-associated vasculitis.

  Yuka Nishibata, Mayu Nonokawa, Yuto Tamura, Rio Higashi, Ku Suzuki, Hideyuki Hayashi, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu

  Clinical and experimental rheumatology 40 (4) 691 - 704 2022/02/04 

  OBJECTIVES: Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is sometimes complicated by anti-glomerular basement membrane (GBM) disease. Proteases, including elastase, released from neutrophils activated by ANCA are implicated in the pathogenesis of AAV. Epitopes of anti-GBM antibody exist in the α3-subunit non-collagenous (NC1) domain of collagen type IV [Col (IV)]. This region, called α3(IV)NC1, is structurally cryptic. This study aimed to determine the production mechanism of anti-GBM antibody in AAV. METHODS: We first examined whether α3(IV)NC1 could be revealed by the digestion of formalin-fixed, paraffin-embedded (FFPE) normal kidney sections and Col (IV) by proteases, including neutrophil elastase, using immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA). Next, the reveal of α3(IV)NC1 and the infiltration of CD11c+ macrophages in the affected kidneys were evaluated by IHC and immunofluorescent staining using FFPE sections. Finally, the production of anti-GBM antibody in AAV rats was determined by ELISA. RESULTS: α3(IV)NC1 was revealed by the digestion of FFPE normal kidney sections and Col (IV) by proteases. Although the reveal of α3(IV)NC1 was observed in sclerotic glomeruli regardless of causative diseases, CD11c+ macrophages near α3(IV)NC1 were characteristics of AAV. Anti-GBM antibody was produced subsequent to ANCA in some AAV rats. IHC demonstrated the reveal of α3(IV)NC1 in affected renal tissues and the infiltration of CD11c+ macrophages around the sites. CONCLUSIONS: The collective findings suggest that, in AAV, proteases released from neutrophils activated by ANCA digest Col (IV) and result in the reveal of α3(IV)NC1, CD11c+ macrophages present GBM epitopes, and then the host's immune system produce anti-GBM antibody.
• 結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の人工知能による鑑別

  柏 航, 加藤 千恵次, 西端 友香, 益田 紗季子, 外丸 詩野, 川上 民裕, 石津 明洋

  脈管学 (一社)日本脈管学会 62 (2) 8 - 8 0387-1126 2022/02
• Involvement of neutrophil extracellular traps in the pathogenesis of glomerulonephritis in a case of systemic lupus erythematosus and antineutrophil cytoplasmic antibody-associated vasculitis overlap syndrome.

  Arisa Senda, Ryutaro Sasai, Kurumi Kato, Yuka Nishibata, Sakiko Masuda, Akihiro Ishizu, Noriko Takahara

  CEN case reports 11 (3) 339 - 346 2192-4449 2022/01/13 

  Systemic lupus erythematosus (SLE) and antineutrophil cytoplasmic antibody-associated vasculitis (AAV) are autoimmune diseases that often cause rapidly progressive glomerulonephritis, with neutrophil extracellular traps (NETs) involved in their pathogenesis. However, the involvement of NETs in the renal damage caused by SLE/AAV overlap syndrome has not been clarified yet. In this study, we detected renal deposition of NETs in a patient with SLE/AAV overlap syndrome. In addition, a significantly increased level of NET-inducing activity was observed in the patient's serum, which improved with treatment. On the other hand, a markedly lower level of NET degradation was observed in the patient's serum as compared to healthy subjects' sera, without any posttreatment changes. These findings suggest that NETs may play a role in the pathogenesis of renal injury associated with SLE/AAV overlap syndrome.
• Neutrophil fixation protocols suitable for substrates to detect anti-neutrophil cytoplasmic antibodies by indirect immunofluorescence.

  Yuka Nishibata, Shun Matsuzawa, Yosuke Satomura, Takeshi Ohtsuka, Motoki Kuhara, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu

  Pathology, research and practice 228 153661 - 153661 0344-0338 2021/12 

  Anti-neutrophil cytoplasmic antibodies (ANCAs) are autoantibodies that recognize neutrophil cytoplasmic antigens. The major ANCA antigens are myeloperoxidase and proteinase 3. Necrotizing small vessel vasculitis accompanied by ANCA production is called ANCA-associated vasculitis (AAV). In addition to AAV, ANCA is sometimes produced in patients with connective tissue diseases, such as systemic lupus erythematosus, and inflammatory bowel diseases. Indirect immunofluorescence (IIF) and enzyme immunoassay (EIA) have been used to detect ANCAs. Recently, the accuracy of EIA has improved and it has become the gold standard for ANCA detection. However, IIF does not lose its role in ANCA detection because EIA cannot detect ANCAs that recognize antigens other than those coated on the plate. For IIF, neutrophil substrates prepared with two different fixations, namely, ethanol fixation and formalin fixation, are used. There is a recommended protocol for ethanol fixation but not for formalin fixation. This study prepared neutrophil substrates according to the recommended protocol for ethanol fixation and protocols in the literature and original protocols for formalin fixation and then examined ANCA specificity and how storage period would influence the number of cells, antigen distribution, and antigenicity of the substrates. As a result, the number of cells and antigen distribution did not change after storage for up to 2 months regardless of fixation protocols, whereas a time-dependent decline in ANCA antigenicity and a fixation protocol-dependent difference in ANCA specificity were observed. How neutrophils are fixed on the glass slide needs to be checked upon evaluation of ANCAs by IIF.
• Spontaneously regressed granulomatosis with polyangiitis: A case report.

  Hiroki Ota, Chisa Sato, Akira Igarashi, Sumito Inoue, Sakiko Masuda, Akihiro Ishizu, Masafumi Watanabe

  Respiratory investigation 59 (3) 372 - 376 2212-5345 2021/05 [Refereed]
   

  A 71-year-old woman presented with chest pain, cough, and back pain. A chest roentgenogram showed multiple nodular shadows in both lungs. She was diagnosed with granulomatosis with polyangiitis (GPA). The multiple nodular shadows in both lungs regressed spontaneously in a few months. There are few reports of spontaneous regression of GPA, and the underlying mechanism is unclear. Neutrophil extracellular traps (NETs) have been recently shown to be involved in GPA. NETs may also be related to the natural regression of GPA.
• Anti-phosphatidylserine/prothrombin complex antibodies in patients with cutaneous vasculitis: Possible involvement in the pathogenesis.

  Tamihiro Kawakami, Yuto Tamura, Yupeng Dong, Miku Yoshinari, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu

  The Journal of dermatology 48 (5) 703 - 706 2021/05 [Refereed]
   

  We assessed the IgG and IgM prevalence of anti-phosphatidylserine/prothrombin complex (aPS/PT) antibodies (Abs) in patients with vasculitis using a novel commercial ELISA kit. To examine whether aPS/PT Abs were involved in the pathogenesis of cutaneous vasculitis, inbred wild-type rats were intravenously administered with a rat IgM class aPS/PT monoclonal Ab established previously or with rat immunoglobulins as controls. To express PS on the surface of vascular endothelium, these rats were given a subcutaneous injection of cell-free histones in advance. Serum IgM aPS/PT Ab levels were elevated in patients with systemic vasculitis with skin involvement and cutaneous arteritis compared to those in patients with systemic vasculitis without skin involvement and healthy controls. There was no significant difference in the serum levels of IgG aPS/PT Abs between the patients and healthy controls. Correspondingly, inbred wild-type rats intravenously administered with the aPS/PT monoclonal IgM Ab after appropriate priming-subcutaneous histone injection developed cutaneous vasculitis. Some rats given rat IgM instead of the aPS/PT monoclonal Ab also developed cutaneous vasculitis, whereas vasculitis did not occur in rats given IgG or only priming by histones. We suggested that IgM aPS/PT Abs could be involved in the pathogenesis of cutaneous vasculitis based on these findings.
• RNase in the saliva can affect the detection of severe acute respiratory syndrome coronavirus 2 by real-time one-step polymerase chain reaction using saliva samples.

  Yuka Nishibata, Shota Koshimoto, Kenta Ogaki, Erika Ishikawa, Kosuke Wada, Miku Yoshinari, Yuto Tamura, Ryo Uozumi, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu

  Pathology, research and practice 220 153381 - 153381 0344-0338 2021/04 [Refereed]
   

  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.
• 人工知能による結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の鑑別

  新海 隼人, 加藤 千恵次, 川上 民裕, 高橋 啓, 西端 友香, 益田 紗季子, 田中 敏, 外丸 詩野, 石津 明洋

  脈管学 (一社)日本脈管学会 61 (1) 1 - 2 0387-1126 2021/01
• Fluvastatin prevents the development of arthritis in env-pX rats via up-regulation of Rho GTPase-activating protein 12.

  Shun Tanimura, Mutsumi Nishida, Tatsunori Horie, Tamotsu Kamishima, Hitomi Matsumoto, Yutaka Morimura, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu

  Experimental and molecular pathology 115 104454 - 104454 0014-4800 2020/08 [Refereed]
   

  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.
• Association of neutrophil extracellular traps with the development of idiopathic osteonecrosis of the femoral head.

  Mayu Nonokawa, Tomohiro Shimizu, Miku Yoshinari, Yamato Hashimoto, Yusuke Nakamura, Daisuke Takahashi, Tsuyoshi Asano, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Norimasa Iwasaki, Akihiro Ishizu

  The American journal of pathology 190 (11) 2282 - 2289 0002-9440 2020/07/20 [Refereed][Not invited]
   

  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 contribute 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 (OA) were assessed for existence of NET-forming neutrophils by immunofluorescent staining. As a result, NET-forming neutrophils were present in small vessels surrounding the femoral head of patients with idiopathic ONFH but not OA. 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 (HIF-1α). NET-forming neutrophils circulated into the tissue around the femoral head, and HIF-1α expression in the tissue was higher compared to 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 intravenous injection of NET-forming neutrophils. The collective findings suggest that NETs are possibly associated with the development of idiopathic ONFH.
• 抗GBM抗体が認識するエピトープの表出に関する検討

  西端 友香, 野々川 茉佑, 益田 紗季子, 中沢 大悟, 田中 敏, 外丸 詩野, 石津 明洋

  脈管学 (一社)日本脈管学会 60 (2) 19 - 19 0387-1126 2020/02
• Relationship between lysosomal-associated membrane protein-2 and anti-phosphatidylserine/prothrombin complex antibody in the pathogenesis of cutaneous vasculitis.

  Tamihiro Kawakami, Ayaka Kikuchi, Chie Miyabe, Takaharu Ikeda, Sora Takeuchi, Yuto Tamura, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu

  Clinical and experimental rheumatology 0392-856X 2020/01/27 [Refereed][Not invited]
   

  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.
• Native myeloperoxidase is required to make the experimental vasculitis model.

  Mayu Nonokawa, Ku Suzuki, Hideyuki Hayashi, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu

  Arthritis research & therapy 21 (1) 296 - 296 1478-6354 2019/12/21 [Refereed][Not invited]
  
• Detection of Increased Vascular Signal in Arthritis-Prone Rats Without Joint Swelling Using Superb Microvascular Imaging Ultrasonography.

  Horie T, Nishida M, Tanimura S, Kamishima T, Tamai E, Morimura Y, Nishibata Y, Masuda S, Nakazawa D, Tomaru U, Atsumi T, Ishizu A

  Ultrasound in medicine & biology 45 (8) 2086 - 2093 0301-5629 2019/08 [Refereed][Not invited]
   

  This study aimed to determine whether ultrasonography (US) can detect increased vascular signal in the synovial tissue before overt synovitis in rheumatoid arthritis (RA). Env-pX rats that spontaneously develop RA-like synovitis were used. Ankle joints of 15 pre-morbid env-pX rats were observed with power Doppler and superb microvascular imaging (SMI) using an ultrahigh-frequency (8-24 MHz) probe. Signal values were counted as the number of pixels. The total number of vessels and vessel area in the synovial tissue were histologically evaluated. Dilated vessels were determined from the mean value of synovial vessels in three wild-type rats. In all env-pX rats, apparent synovial proliferation was not observed. However, vasodilation was evident. Only SMI values were significantly correlated with the number of dilated vessels (r = 0.585, p = 0.022) but not with the total number of vessels. US with SMI using ultrahigh-frequency probe can detect increased vascular signal in the synovial tissue of arthritis-prone rats.
• The presence of anti-neutrophil extracellular trap antibody in patients with microscopic polyangiitis.

  Hattanda F, Nakazawa D, Watanabe-Kusunoki K, Kusunoki Y, Shida H, Masuda S, Nishio S, Tomaru U, Atsumi T, Ishizu A

  Rheumatology (Oxford, England) 58 (7) 1293 - 1298 1462-0324 2019/07 [Refereed][Not invited]
   

  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.
• Formation and Disordered Degradation of Neutrophil Extracellular Traps in Necrotizing Lesions of Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis.

  Masuda S, Nonokawa M, Futamata E, Nishibata Y, Iwasaki S, Tsuji T, Hatanaka Y, Nakazawa D, Tanaka S, Tomaru U, Kawakami T, Atsumi T, Ishizu A

  The American journal of pathology 189 (4) 839 - 846 0002-9440 2019/04 [Refereed][Not invited]
   

  Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is characterized by the production of ANCAs and systemic necrotizing vasculitis in small vessels. Disordered regulation of neutrophil extracellular traps (NETs) is critically involved in the pathogenesis of AAV. NETs are web-like DNA decorated with antimicrobial proteins; they are extruded from activated neutrophils. The principal degradation factor of NETs in vivo is DNase I; however, NETs resistant to DNase I can persist in tissues and can lead to the production of ANCAs. Deposition of NETs has been demonstrated in glomerular crescents and necrotizing vasculitis in AAV. Here, the amount of NETs in formalin-fixed, paraffin-embedded tissue sections was examined, and the results for AAV were compared with the results for diseases that should be distinguished from AAV. NETs were more abundant in necrotizing vasculitis of AAV than in non-ANCA-associated vasculitis, or in granulomatous angiitis. Pulmonary granulomas in AAV and non-ANCA-associated diseases were further studied. The amount of NETs was significantly greater in necrotizing granulomas of AAV than in granulomas of sarcoidosis without necrosis. Although NETs were formed in necrotizing granulomas of tuberculosis equivalently to those formed in AAV, they were more susceptible to degradation by DNase I than were NETs in AAV. The formation and disordered degradation of NETs in necrotizing lesions are characteristics of AAV and are possibly related to its pathogenesis.
• Epitope recognized by anti-glomerular basement membrane (GBM) antibody in a patient with repeated relapse of anti-GBM disease.

  Nishibata Y, Masuda S, Nakazawa D, Tanaka S, Tomaru U, Nergui M, Jia X, Cui Z, Zhao MH, Nakabayashi K, Ishizu A

  Experimental and molecular pathology 107 165 - 170 0014-4800 2019/04 [Refereed][Not invited]
   

  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.
• Pharmaceutical immunoglobulins reduce neutrophil extracellular trap formation and ameliorate the development of MPO-ANCA-associated vasculitis.

  Uozumi R, Iguchi R, Masuda S, Nishibata Y, Nakazawa D, Tomaru U, Ishizu A

  Modern rheumatology 30 (3) 1 - 7 1439-7595 2019/04 [Refereed][Not invited]
   

  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.
• HIGH DOSE IMMUNOGLOBULINS CAN INHIBIT NEUTROPHIL EXTRACELLULAR TRAP FORMATION, EVENTUALLY PREVENT THE DEVELOPMENT OF MPO-ANCA-ASSOCIATED VASCULITIS

  Ryo Uozumi, Sakiko Masuda, Yuka Nishibata, Shun Tanimura, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu

  RHEUMATOLOGY 58 1462-0324 2019/03
• 抗糸球体基底膜抗体病(抗GBM病)の頻回再発症例の抗GBMが認識するエピトープ

  西端 友香, 東 里緒, 益田 紗季子, 中沢 大悟, 田中 敏, 外丸 詩野, 中林 公正, 石津 明洋

  脈管学 (一社)日本脈管学会 59 (3) 19 - 19 0387-1126 2019/03
• ヒストンは好中球細胞上にLAMP2を表出し、抗LAMP2抗体と連携して皮膚小血管炎の発症機序に関与している

  川上 民裕, 竹内 そら, 菊池 彩翔, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋

  脈管学 (一社)日本脈管学会 59 (3) 22 - 22 0387-1126 2019/03
• Author Correction: Pathogenesis and therapeutic interventions for ANCA-associated vasculitis.

  Daigo Nakazawa, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu

  Nature reviews. Rheumatology 15 (2) 123 - 123 2019/02 

  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.
• Pathogenesis and therapeutic interventions for ANCA-associated vasculitis.

  Nakazawa D, Masuda S, Tomaru U, Ishizu A

  Nature reviews. Rheumatology 15 (2) 91 - 101 1759-4790 2019/02 [Refereed][Not invited]
   

  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.
• EFFECTS OF RECOMBINANT THROMBOMODULIN ON EXPERIMENTAL AUTOIMMUNE VASCULITIS VIA THE INHIBITION OF NEUTROPHIL EXTRACELLULAR TRAPS

  Kanako Watanabe, Daigo Nakazawa, Yoshihiro Kusunoki, Fumihiko Hattanda, Saori Nishio, Sakiko Masuda, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu

  NEPHROLOGY DIALYSIS TRANSPLANTATION 33 34 - 34 0931-0509 2018/05
• ANCA関連血管炎の壊死性病変部におけるNETsの存在と病的意義

  益田 紗季子, 野々川 茉佑, 西端 友香, 岩崎 沙理, 辻 隆裕, 田中 敏, 外丸 詩野, 川上 民裕, 石津 明洋

  脈管学 (一社)日本脈管学会 58 (3) 35 - 35 0387-1126 2018/03
• プロテアーゼによる抗糸球体基底膜抗体病の隔絶抗原の表出

  西端 友香, 植松 千浩, 益田 紗季子, 田中 敏, 外丸 詩野, 石津 明洋

  脈管学 (一社)日本脈管学会 58 (3) 36 - 37 0387-1126 2018/03
• 血管炎の実験動物モデル 抗PSPT抗体は、正常ラットに血栓を発症させる

  川上 民裕, 山田 真衣, 高島 滉平, 西岡 佑介, 西端 友香, 益田 紗季子, 吉田 繁, 外丸 詩野, 石津 明洋

  脈管学 (一社)日本脈管学会 57 (2) 22 - 22 0387-1126 2017/02

Conference Activities & Talks

• ANCA関連血管炎(AAV)の壊死性病変部における好中球細胞外トラップ(NETs)の存在と病的意義  [Not invited]

  益田 紗季子

  日本病理学会  2018
• ANCA関連血管炎(AAV)の壊死性病変部における好中球細胞外トラップ(NETs)の存在と病的意義  [Not invited]

  益田 紗季子

  血管病理研究会  2017
• フローサイトメトリーによる定量的好中球細胞外トラップ測定法  [Not invited]

  益田 紗季子

  日本病理学会  2017
• フローサイトメトリーによる定量的好中球細胞外トラップ測定法  [Not invited]

  益田 紗季子

  MPO研究会  2016
• 初発診断および再発評価に細胞診が有用だった子宮頸部原発小細胞癌の１例  [Not invited]

  益田 紗季子

  日本臨床細胞学会  2014
• ヒト末梢血におけるCD8陽性単球・顆粒球の解析  [Not invited]

  益田 紗季子

  日本病理学会  2010

MISC

• Rising starの集い ANCA関連血管炎におけるNETs形成異常

  益田 紗季子  脈管学  64-  (1)  5  -6  2024/02
• 結腸癌の診断と同時期に発症した抗LAMP-2抗体陽性血管炎の1例

  原田 拓弥, 山下 裕之, 上田 聖, 秋山 優弥, 小林 俊昭, 谷口 舞, 西端 友香, 益田 紗季子, 石津 明洋, 金子 礼志  脈管学  64-  (1)  11  -12  2024/02
• 壊疽性膿皮症の皮膚生検標本を使用したneutrophil extracellular traps(NETs)の検証

  川上 民裕, 横山 華英, 池田 高治, 益田 紗季子, 石津 明洋  日本皮膚科学会雑誌  133-  (4)  713  -714  2023/04
• 皮膚血管炎動物モデルの完成

  川上民裕, 中出一生, 田村宥人, 伊藤吹夕, 西端友香, 益田紗季子, 外丸詩野, 石津明洋  脈管学(Web)  63-  (1)  2023
• 正常ラットに抗PSPT抗体と抗LAMP2抗体の静脈注射により皮膚血管炎の発症に成功した

  川上民裕, 中出一生, 田村宥人, 西端友香, 益田紗季子, 石津明洋, 伊藤吹夕, 外丸詩野  日本皮膚科学会雑誌  133-  (3)  523  -523  2023
• 正常ラットにヒストン皮下注射後,抗ホスファチジルセリン・プロトロンビン複合体抗体と抗リソソーム膜タンパク質2抗体の静脈注射により,皮膚血管炎の発症に成功した

  川上 民裕, 中出 一生, 田村 宥人, 伊藤 吹夕, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋  日本皮膚免疫アレルギー学会総会学術大会プログラム・抄録集  52回-  215  -215  2022/12
• ベーチェット病皮下の血栓性静脈炎におけるNeutrophil Extracellular Trapsの発現

  川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋  アレルギー  71-  (6-7)  870  -870  2022/08
• 結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の人工知能による鑑別

  柏 航, 加藤 千恵次, 西端 友香, 益田 紗季子, 外丸 詩野, 川上 民裕, 石津 明洋  脈管学  62-  (2)  8  -8  2022/02
• 抗ホスファチジルセリン・プロトロンビン複合体抗体による皮膚血管炎動物モデルの完成

  川上 民裕, 董 宇鵬, 田村 宥人, 吉成 未来, 西端 友香, 益田 紗季子, 石津 明洋, 外丸 詩野  日本皮膚科学会雑誌  132-  (1)  91  -91  2022/01
• 【新しい手法を駆使した腎臓病研究の最前線】糸球体疾患 ANCAとNETs

  石津 明洋, 益田 紗季子, 西端 友香  腎と透析  91-  (5)  851  -855  2021/11
• 【循環器II-血管・血管腫・弁膜疾患-】小型血管の血管炎

  石津 明洋, 益田 紗季子, 西端 友香  病理と臨床  39-  (11)  1116  -1122  2021/11
• 免疫疾患の形態病理学的理解 ヒストン皮下注射と抗ホスファチジルセリン・プロトロンビン複合体抗体静脈注射から完成した皮膚血管炎の動物モデル

  川上 民裕, 田村 宥人, 董 宇鵬, 吉成 未来, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋  日本臨床免疫学会総会プログラム・抄録集  49回-  58  -58  2021/10
• 膠原病・自己免疫疾患 皮膚血管炎動物モデルの完成と抗ホスファチジルセリン・プロトロンビン複合体抗体

  川上 民裕, 田村 宥人, 董 宇鵬, 吉成 未来, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋  アレルギー  70-  (6-7)  809  -809  2021/08
• 多発血管炎性肉芽腫症(GPA)の自然退縮における,好中球細胞外トラップ(NETs)分解について 当院で経験した1症例を通して

  太田 啓貴, 佐藤 千紗, 五十嵐 朗, 邨野 浩義, 梁 秀鼎, 町田 浩祥, 佐藤 建人, 中野 寛之, 根本 貴子, 西脇 道子, 木村 友美, 山内 啓子, 佐藤 正道, 井上 純人, 渡辺 昌文, 益田 紗季子, 石津 明洋  日本呼吸器学会誌  10-  (増刊)  241  -241  2021/04
• 人工知能による結節性多発動脈炎と皮膚動脈炎の皮膚生検画像の鑑別

  新海 隼人, 加藤 千恵次, 川上 民裕, 高橋 啓, 西端 友香, 益田 紗季子, 田中 敏, 外丸 詩野, 石津 明洋  脈管学  61-  (1)  1  -2  2021/01
• myosin light chain6を認議する抗好中球細胞外トラップ(NETs)抗体はNETs分解阻害活性を持つ

  西端友香, 益田紗季子, 外丸詩野, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  65th-  407  -407  2021
• Effects of hypoxia on neutrophil extracellular trap formation

  益田紗季子, 西端友香, 田中敏, 外丸詩野, 辻野一三, 石津明洋  日本病理学会会誌  110-  (1)  246  -246  2021
• Degradation of neutrophil extracellular traps in the spontaneous regression of granulomatosis with polyangiitis: a case report

  太田啓貴, 佐藤千紗, 五十嵐朗, 邨野浩義, 梁秀鼎, 町田浩祥, 佐藤建人, 中野寛之, 根本貴子, 西脇道子, 木村友美, 山内啓子, 佐藤正道, 井上純人, 渡辺昌文, 益田紗季子, 石津明洋  日本呼吸器学会誌(Web)  10-  2021
• 腎生検組織におけるNeutrophil Extracellular Traps(NETs)の検討

  岩崎 沙理, 益田 紗季子, 石津 明洋, 大塚 拓也, 牧田 啓史, 深澤 雄一郎, 辻 隆裕  日本病理学会会誌  109-  (1)  435  -435  2020/03
• Recombinant thrombomodulin ameliorates autoimmune vasculitis via immune response regulation and tissue injury protection.

  Kanako Watanabe-Kusunoki, Daigo Nakazawa, Yoshihiro Kusunoki, Takashi Kudo, Fumihiko Hattanda, Saori Nishio, Sakiko Masuda, Utano Tomaru, Takeshi Kondo, Tatsuya Atsumi, Akihiro Ishizu  Journal of autoimmunity  108-  102390  -102390  2020/03  [Not refereed][Not invited]
   

  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.
• 抗GBM抗体が認識するエピトープの表出に関する検討

  西端友香, 野々川茉佑, 益田紗季子, 中沢大悟, 田中敏, 外丸詩野, 石津明洋  脈管学(Web)  60-  (2)  2020
• ANCA関連血管炎の病因研究 ANCA関連血管炎における好中球細胞外トラップ

  魚住諒, 魚住諒, 益田紗季子, 石津明洋  炎症と免疫  28-  (4)  279  -283  2020  

  好中球細胞外トラップ(NETs)は、活性化した好中球から細胞外に放出されるDNA線維であり、好中球細胞質の抗菌タンパクで装飾されている。NETsは生体防御に不可欠な自然免疫機構であるが、過剰なNETsは血管内皮障害の原因になるとともに、ANCA産生の原因ともなる。ANCA関連血管炎では、ANCAによるNETsの誘導、セマフォリン4Dを介したNETs抑制機構の不全、NETsの生理的な分解因子であるDNase Iの活性低下、不明の機序によるNETs自体のDNase I抵抗性獲得などのため、NETsが体内に蓄積しやすい状況が生じている。(著者抄録)
• 特発性大腿骨頭壊死症の発生における好中球細胞外トラップの関与

  清水智弘, 野々川茉佑, 西端友香, 益田紗季子, 高橋大介, 浅野毅, 田中敏, 外丸詩野, 岩崎倫政, 石津明洋  日本整形外科学会雑誌  94-  (8)  S1801  -S1801  2020
• Formation of degradation-resistant NETs in pneumonia patients with lung disease

  益田紗季子, 石橋美郷, 加藤くるみ, 西端友香, 田中敏, 外丸詩野, 辻野一三, 石津明洋  日本病理学会会誌  109-  (1)  308  -308  2020
• Detection of Neutrophil Extracellular Traps in kidney biopsy specimen

  岩崎沙理, 益田紗季子, 石津明洋, 大塚拓也, 牧田啓史, 深澤雄一郎, 辻隆裕  日本病理学会会誌  109-  (1)  2020
• 抗好中球細胞質抗体(ANCA)に続き抗糸球体基底膜(GBM)抗体が産生されるメカニズム

  西端友香, 益田紗季子, 中沢大悟, 外丸詩野, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  64th (CD-ROM)-  507  -507  2020
• 薬剤起因性自己免疫疾患におけるNETsの関与

  益田紗季子, 石津明洋  月刊リウマチ科  61-  (1)  64  -68  2019
• 特発性大腿骨頭壊死症の発生における好中球細胞外トラップの関与

  野々川茉佑, 西端友香, 益田紗季子, 石津明洋, 清水智弘, 高橋大介, 浅野毅, 岩崎倫政, 田中敏, 外丸詩野  北海道医学雑誌  94-  (1)  59  -59  2019
• ANCA関連血管炎(AAV)の壊死性病変部における好中球細胞外トラップ(NETs)の存在と病的意義

  益田紗季子, 野々川茉佑, 西端友香, 岩崎沙理, 辻隆裕, 田中敏, 外丸詩野, 川上民裕, 石津明洋  日本病理学会会誌  108-  (1)  327  -327  2019
• 抗糸球体基底膜抗体病(抗GBM病)の頻回再発症例の抗GBMが認識するエピトープ

  西端友香, 東里緒, 益田紗季子, 中沢大悟, 田中敏, 外丸詩野, 中林公正, 石津明洋  脈管学(Web)  59-  (3)  2019
• シクロフィリンDをターゲットとしたANCA関連壊死性血管炎に対する新規治療薬の開発

  工藤孝司, 中沢大悟, 白鳥里佳, 楠加奈子, 西尾妙織, 益田紗季子, 外丸詩野, 石津明洋, 渥美達也  日本臨床免疫学会総会プログラム・抄録集  47th-  134  -134  2019
• 抗GBM抗体が認識するエピトープの表出に関する検討

  西端友香, 東里緒, 益田紗季子, 中沢大悟, 田中敏, 外丸詩野, 石津明洋  日本病理学会会誌  108-  (1)  298  -298  2019
• ANCA関連血管炎の壊死性病変部における好中球細胞外トラップの存在と病的意義

  益田紗季子, 益田紗季子, 西端友香, 中沢大悟, 外丸詩野, 川上民裕, 渥美達也, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  63rd-  2019
• 皮膚血管炎の発症機序における抗リソソーム膜タンパク2抗体(抗LAMP2抗体)と抗ホスファチジルセリン・プロトロンビン複合体抗体(抗PSPT抗体)の役割

  川上民裕, 宮部千恵, 池田高治, 菊池彩花, 西端友香, 益田紗季子, 石津明洋, 中沢大悟, 外丸詩野  日本皮膚科学会雑誌  129-  (12)  2533  -2533  2019
• ヒストンは好中球細胞上にLAMP2を表出し,抗LAMP2抗体と連携して皮膚小血管炎の発症機序に関与している

  川上民裕, 竹内そら, 菊池彩翔, 西端友香, 益田紗季子, 外丸詩野, 石津明洋  脈管学(Web)  59-  (3)  2019
• ANCA関連血管炎(AAV)の壊死性病変部における好中球細胞外トラップ(NETs)の存在と病的意義

  益田紗季子, 野々川茉佑, 西端友香, 岩崎沙理, 辻隆裕, 田中敏, 外丸詩野, 川上民裕, 石津明洋  日本病理学会会誌  107-  (1)  330  -330  2018/04/26  [Not refereed][Not invited]
  
• 免疫グロブリン大量静注療法(IVIG)は好中球細胞外トラップ(NETs)の抑制によりMPO‐ANCA関連血管炎を抑制する

  魚住諒, 魚住諒, 井口理彩, 益田紗季子, 西端友香, 谷村瞬, 中沢大悟, 外丸詩野, 石津明洋  日本病理学会会誌  107-  (1)  331  -331  2018/04/26  [Not refereed][Not invited]
  
• 免疫グロブリン大量静注療法(IVIG)は好中球細胞外トラップ(NETs)の抑制によりMPO-ANCA関連血管炎を抑制する

  魚住 諒, 井口 理彩, 益田 紗季子, 西端 友香, 谷村 瞬, 中沢 大悟, 外丸 詩野, 石津 明洋  日本病理学会会誌  107-  (1)  331  -331  2018/04  [Not refereed][Not invited]
  
• 免疫グロブリン製剤は好中球細胞外トラップ(NETs)の形成抑制を介してMPO‐ANCA関連血管炎(MPO‐AAV)の発症を抑制する

  魚住諒, 魚住諒, 益田紗季子, 西端友香, 谷村瞬, 中沢大悟, 外丸詩野, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  62nd-  433  -433  2018/03/20  [Not refereed][Not invited]
  
• スタチンの関節炎抑制効果とその機序の解明

  谷村瞬, 西田睦, 神島保, 西端友香, 益田紗季子, 中沢大悟, 外丸詩野, 渥美達也, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  62nd-  778  -778  2018/03/20  [Not refereed][Not invited]
  
• 超高周波プローブを用いたSuperb Micro‐vascular Imagingによる超音波検査でのラット足関節の早期関節炎診断

  西田睦, 谷村瞬, 神島保, 西端友香, 益田紗季子, 中沢大悟, 外丸詩野, 渥美達也, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  62nd-  485  -485  2018/03/20  [Not refereed][Not invited]
  
• 血管炎1:血管炎のバイオマーカー・病態解析 免疫グロブリン製剤は好中球細胞外トラップ(NETs)の形成抑制を介してMPO-ANCA関連血管炎(MPO-AAV)の発症を抑制する

  魚住 諒, 益田 紗季子, 西端 友香, 谷村 瞬, 中沢 大悟, 外丸 詩野, 石津 明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  62回-  433  -433  2018/03  [Not refereed][Not invited]
  
• リウマチ性疾患の画像2:関節エコーその他 超高周波プローブを用いたSuperb Micro-vascular Imagingによる超音波検査でのラット足関節の早期関節炎診断

  西田 睦, 谷村 瞬, 神島 保, 西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 渥美 達也, 石津 明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  62回-  485  -485  2018/03  [Not refereed][Not invited]
  
• スタチンの関節炎抑制効果とその機序の解明

  谷村 瞬, 西田 睦, 神島 保, 西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 渥美 達也, 石津 明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  62回-  778  -778  2018/03  [Not refereed][Not invited]
  
• 抗好中球細胞質抗体(ANCA)と好中球細胞外トラップ(NETs)

  益田紗季子, 石津明洋  月刊リウマチ科  59-  (2)  200‐205  -205  2018/02/28  [Not refereed][Not invited]
  
• BDNFはHUVECにおけるangiogeninの分泌と核移行を誘導する

  森綾子, 西岡祐介, 山田真衣, 西端友香, 益田紗季子, 外丸詩野, 本間直幸, 森山隆則, 石津明洋  日本血管生物医学会学術集会プログラム・抄録集  26th-  2018
• スタチンの関節炎抑制効果と機序の検討

  谷村瞬, 渥美達也, 西田睦, 堀江達則, 神島保, 玉井絵里香, 森村豊, 西端友香, 益田紗季子, 石津明洋, 外丸詩野  北海道医学雑誌  93-  (2)  121  -122  2018
• プロテアーゼによる抗糸球体基底膜抗体病の隔絶抗原の表出

  西端友香, 植松千浩, 益田紗季子, 田中敏, 外丸詩野, 石津明洋  脈管学(Web)  58-  (3)  2018
• ANCA関連血管炎の壊死性病変部におけるNETsの存在と病的意義

  益田紗季子, 野々川茉佑, 西端友香, 岩崎沙理, 辻隆裕, 田中敏, 外丸詩野, 川上民裕, 石津明洋  脈管学(Web)  58-  (3)  2018
• スタチン製剤の関節炎抑制効果と機序の検討

  谷村瞬, 渥美達也, 西田睦, 堀江達則, 神島保, 齋藤克己, 西端友香, 益田紗季子, 石津明洋, 外丸詩野  北海道医学雑誌  92-  (2)  105  -105  2017/11/01  [Not refereed][Not invited]
  
• 自己免疫疾患とNETosis

  楠由宏, 益田紗季子, 外丸詩野, 石津明洋  月刊リウマチ科  57-  (4)  437‐442  -442  2017/04/28  [Not refereed][Not invited]
  
• フローサイトメトリーによる定量的好中球細胞外トラップ測定法

  益田紗季子, 西端友香, 松尾淳司, 外丸詩野, 石津明洋  日本病理学会会誌  106-  (1)  350  -350  2017/03/24  [Not refereed][Not invited]
  
• 抗PSPT抗体による抗リン脂質抗体症候群動物モデルの完成

  川上民裕, 石津明洋, 益田紗季子, 外丸詩野  日本リウマチ学会総会・学術集会プログラム・抄録集  61st-  605  -605  2017/03/18  [Not refereed][Not invited]
  
• ANCA関連血管炎(AAV)における抗NETs抗体の存在と病的意義

  八反田文彦, 楠由宏, 志田玄貴, 中沢大悟, 西尾妙織, 益田紗季子, 外丸詩野, 渥美達也, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  61st-  431  -431  2017/03/18  [Not refereed][Not invited]
  
• フローサイトメトリーによる定量的好中球細胞外トラップ測定法

  益田 紗季子, 西端 友香, 松尾 淳司, 外丸 詩野, 石津 明洋  日本病理学会会誌  106-  (1)  350  -350  2017/03  [Not refereed][Not invited]
  
• _7 RAT TYPE II NKT CELL CLONE PATHOGENIC FOR SMALL VESSEL VASCULITIS RECOGNIZES STEROL CARRIER PROTEIN 2

  Yusuke Nishioka, Madoka Yamaguchi, Ai Kawakami, Maya Munehiro, Sakiko Masuda, Utano Tomaru, Akihiro Ishizu  RHEUMATOLOGY  56-  30  -31  2017/03  [Not refereed][Not invited]
  
• THE EFFECT OF PEPTIDYLARGININE DEIMINASE INHIBITOR ON NET FORMATION AND MPO-ANCA PRODUCTION IN MOUSE MODEL

  Yoshihiro Kusunoki, Daigo Nakazawa, Haruki Shida, Fumihiko Hattanda, Arina Miyoshi, Sakiko Masuda, Saori Nishio, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu  RHEUMATOLOGY  56-  112  -113  2017/03  [Not refereed][Not invited]
  
• ESTABLISHMENT OF ANTI-RAT PHOSPHATIDYLSERINE/PROTHROMBIN MONOCLONAL ANTIBODIES AND A THROMBOTIC RAT MODEL INDUCED BY INTRAVENOUS INJECTION OF THE ANTIBODY

  Mai Yamada, Tamihiro Kawakami, Kohei Takashima, Yusuke Nishioka, Yuka Nishibata, Sakiko Masuda, Shigeru Yoshida, Akihiro Ishizu  RHEUMATOLOGY  56-  2017/03  [Not refereed][Not invited]
  
• 抗PSPT抗体は,正常ラットに血栓を発症させる

  川上民裕, 山田真衣, 高島滉平, 西岡佑介, 西端友香, 益田紗季子, 吉田繁, 外丸詩野, 石津明洋  脈管学(Web)  57-  (2)  22(J‐STAGE)  2017  [Not refereed][Not invited]
  
• MPO‐ANCA関連血管炎におけるNETs‐ANCA悪循環

  志田玄貴, 八反田文彦, 三次有奈, 楠由宏, 中沢大悟, 佐藤遥, 橋本展洋, 林晃正, 益田紗季子, 石津明洋, 外丸詩野  北海道医学雑誌  91-  (2)  85‐86  -86  2016/11/01  [Not refereed][Not invited]
  
• ANCA関連血管炎―最近の話題―1 好中球細胞外トラップ

  益田紗季子, 石津明洋  月刊アレルギーの臨床  36-  (485)  419‐423  -423  2016/05/20  [Not refereed][Not invited]
   

  ANCA関連血管炎において、ANCAは主要な病原因子である。このANCAの産生およびANCAを介した血管障害メカニズムに、好中球細胞外トラップ(Neutrophil Extracellular Traps:NETs)が関与することが明らかとなってきた。感染刺激を受けた好中球は活性化され、NETsを放出して細胞死に至る。NETsは対感染防御において不可欠であるものの、その制御異常はANCAの産生原因となる。また、産生されたANCAにはNETs誘導活性があり、NETsには血管障害活性がある。NETsはANCA関連血管炎の病因ならびに病態形成に重要な役割を果たしている。(著者抄録)
• 自己血管内皮細胞反応性小型血管炎惹起性type II NKT細胞が認識する分子の同定

  西岡佑介, 山口まどか, 川上愛, 宗廣真矢, 山田真衣, 益田紗季子, 外丸詩野, 石津明洋  日本病理学会会誌  105-  (1)  353  -353  2016/04/12  [Not refereed][Not invited]
  
• 抗ラクトフェリン抗体は好酸球性多発血管炎性肉芽腫症において好中球細胞外トラップの形成を促進し,疾患活動性に関与する

  志田玄貴, 中沢大悟, 八反田文彦, 楠由宏, 益田紗季子, 外丸詩野, 川上民裕, 渥美達也, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  60th-  479  -479  2016/03/18  [Not refereed][Not invited]
  
• Peptigylarginine deiminase 4阻害薬は好中球細胞外トラップの形成阻害を介してMPO‐ANCA産生を抑制する

  楠由宏, 中沢大悟, 志田玄貴, 八反田文彦, 益田紗季子, 外丸詩野, 西尾妙織, 渥美達也, 石津明洋  日本リウマチ学会総会・学術集会プログラム・抄録集  60th-  479  -479  2016/03/18  [Not refereed][Not invited]
  
• Glandular papilloma(columnar cell papilloma)の1例

  KATO TAKASHI, MASUDA SAKIKO, YOSHIOKA ASUKA, HIRAO TOMOMI, HIDA IKUHARU, ICHIHARA MAKOTO, GOTODA YUKO, MURAOKA SHUNJI  日本臨床細胞学会雑誌  53-  (Suppl.2)  584  -584  2014/10/10  [Not refereed][Not invited]
  
• 初発診断および再発評価に細胞診が有用だった子宮頸部原発小細胞癌の1例

  MASUDA SAKIKO, KATO TAKASHI, YOSHIOKA ASUKA, HIRAO TOMOMI, ICHIHARA SHIN, GOTODA YUKO, MURAOKA SHUNJI  日本臨床細胞学会雑誌  53-  (Suppl.1)  266  -266  2014/04/23  [Not refereed][Not invited]
  
• A case report of matrix-producing carcinoma of the breast

  MINOSHIMA ATSUSHI, MASUDA SAKIKO, KATO TAKASHI, YOSHIOKA ASUKA, HIRAO TOMOMI, ICHIHARA SHIN, GOTODA HIROKO, MURAOKA SHUNJI  日本臨床細胞学会雑誌  53-  (1)  41  -46  2014/01/22  [Not refereed][Not invited]
   

  <b><i>Background</i></b> : Matrix-producing carcinoma (MPC) of the breast is a rare subtype of breast cancer, accounting for 0.03 to 0.2% of all breast cancers.<br>We report a case of MPC and its cytopathological features.<br><b><i>Case</i></b> : A 53-year-old female presented with a hard mass in her left breast. Papanicolaou staining of fine-needle-aspiration cytology specimens showed atypical single cells and cell clusters in a pale green-stained myxoid background. There were three types of cell clusters : carcinomatous, chondromatous, and intermediate clusters in a chondroid matrix.<br>Histologically, the tumor had a chondromatous element at the center and a carcinomatous element in the peripheral region. The tumor demonstrated overt carcinoma with direct transition to a cartilaginous stromal matrix without an intervening spindle cell zone or osteoclastic cells ; based on these findings, we diagnosed the tumor as MPC.<br><b><i>Conclusion</i></b> : It is possible to suspect MPC when both carcinomatous and chondromatous cell clusters appear on the same glass slide with transitional images of these cells. Thus, MPC may be suspected clinically based on its characteristic properties.
• 乳腺基質産生癌の一例

  MINOSHIMA ATSUSHI, MASUDA SAKIKO, KATO TAKASHI, YOSHIOKA ASUKA, HIRAO TOMOMI, ICHIHARA MAKOTO, GOTODA YUKO, MURAOKA SHUNJI  日本臨床細胞学会雑誌  52-  (Suppl.1)  197  -197  2013/05/17  [Not refereed][Not invited]
  
• Plasma-dependent, antibody- and Fcγ receptor-mediated translocation of CD8 molecules from T cells to monocytes

  岩崎 沙理, 益田 紗季子, 馬場 智久, 外丸 詩野, 勝俣 一晃, 笠原 正典, 石津 明洋  北海道醫學雜誌 = Acta medica Hokkaidonensia  87-  (2)  68  -68  2012/04/01  [Not refereed][Not invited]
  
• Fcγ受容体を介したtrogocytosisの意義と制御機構の解析

  MASUDA SAKIKO, IWASAKI SARI, SATO JURI, TOMARU UTANO, KASAHARA MASANORI, ISHIZU AKIHIRO  日本病理学会会誌  100-  (1)  316  -316  2011/03/26  [Not refereed][Not invited]
  
• ヒト末梢血に検出されるCD8陽性単球・顆粒球の解析

  MASUDA SAKIKO, IWASAKI SATOSHI, BABA TOMOHISA, KATSUMATA KAZUAKI, TOMARU UTANO, KASAHARA MASANORI, ISHIZU AKIHIRO  日本病理学会会誌  99-  (1)  217  -217  2010/03/26  [Not refereed][Not invited]
  
• ヒト末梢血におけるCD8陽性単球の解析

  IWASAKI SARI, MASUDA SAKIKO, BABA TOMOHISA, KATSUMATA KAZUAKI, TOMARU UTANO, KASAHARA MASANORI, ISHIZU AKIHIRO  臨床病理  57-  (補冊)  136  -136  2009/07/28  [Not refereed][Not invited]
  
• ヒト末梢血に検出されるCD8陽性単球の解析

  IWASAKI SARI, BABA TOMOHISA, MASUDA SAKIKO, KATSUMATA KAZUAKI, TOMARU UTANO, KASAHARA MASANORI, ISHIZU AKIHIRO  日本病理学会会誌  98-  (1)  279  -279  2009/03/20  [Not refereed][Not invited]
  

Research Grants & Projects

• ベーチェット病患者唾液の好中球細胞外トラップ誘導能の低下原因の解明

  日本学術振興会：科学研究費助成事業

  Date (from‐to) : 2022/04 -2025/03 

  Author : 益田 紗季子
• Reveal of the pathogenesis of MPO-ANCA-associated vasculitis and development of novel therapeutic strategies

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2021/04 -2024/03 

  Author : 石津 明洋, 益田 紗季子, 外丸 詩野, 中沢 大悟

   

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

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2019/04 -2022/03 

  Author : Masuda Sakiko

   

  The proteins of DNase I-sensitive NETs or DNase I-resistant NETs were compared by proteomics analysis, and narrowed down 18 protein candidates that are considered to confer DNase I resistance to NETs. When candidate proteins were added during NET induction, the formation of normal NET was suppressed by the five proteins, and the formation of abnormal NET resistant to DNase I became the main component. Addition of an inhibitory antibody to one of these proteins during NETs induction suppressed the formation of normal and abnormal NETs.

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