Cathepsin C inhibition reduces neutrophil serine protease activity and improves activated neutrophil-mediated disorders.
Yuka Nishibata, Suishin Arai, Mai Taniguchi, Issei Nakade, Hodaka Ogawa, Shota Kitano, Yumeka Hosoi, Ayano Shindo, Ryo Nishiyama, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Takafumi Shimizu, William Sinko, Tadashi Nagakura, Yoh Terada, Akihiro Ishizu
Nature communications, 15, 1, 6519, 6519, 2024年08月22日, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Cathepsin C (CatC) is an enzyme which regulates the maturation of neutrophil serine proteases (NSPs) essential for neutrophil activation. Activated neutrophils are key players in the innate immune system, and are also implicated in the etiology of various inflammatory diseases. This study aims to demonstrate a therapeutic potential for CatC inhibitors against disorders in which activated neutrophil-derived neutrophil extracellular traps (NETs) play a significant role. We demonstrate that a CatC inhibitor, MOD06051, dose-dependently suppresses the cellular activity of NSPs, including neutrophil elastase (NE), in vitro. Neutrophils derived from MOD06051-administered rats exhibit significantly lower NE activity and NET-forming ability than controls. Furthermore, MOD06051 dose-dependently ameliorates vasculitis and significantly decreases NETs when administered to a rat model of myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody-associated vasculitis (AAV). These findings suggest that CatC inhibition is a promising strategy to reduce neutrophil activation and improve activated neutrophil-mediated diseases such as MPO-AAV.

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年12月28日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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年11月06日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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, Elsevier BV, 2023年10月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: Anti-glomerular basement membrane (anti-GBM) antibody is essential for the diagnosis of anti-GBM disease. The major epitope consists of the α3 subunits of type IV collagen non-collagenous domain (α 3(IV)NC1). There have been only a few reports of patients false-positive for anti-GBM antibody. CASE REPORT: We experienced an 8-year-old boy who presented with asymptomatic hematuria followed by positivity for anti-GBM antibody as evaluated by a commercially available chemiluminescent enzyme immunoassay (CLEIA). While his condition remained stable other than continuing hematuria, his anti-GBM antibody titer increased. Further examination of another anti-GBM antibody assay (fluoroenzyme immunoassay) showed negative results. Thus, evaluation of the accuracy of his positivity for anti-GBM antibody was required. We conducted the following examinations: A) enzyme-linked immunosorbent assay, B) immunoblotting for recombinant α 1-5(IV)NC1, and C) immunohistochemical analysis of normal kidney tissue sections. Specimens used for the analysis were sera in A and IgG from the patient in B and C, respectively. As a result, no anti-GBM antibody was detected in A. In B, no band specific to α 1-5(IV)NC1 was observed. In C, the kidney tissue was not stained. Taken together, these results led us to judge the positive anti-GBM result in CLEIA of our patient to be a non-specific reaction. CONCLUSION: The commercial assays for anti-GBM antibody can lead to false-positive results. We recommend confirmation of anti-GBM antibody positivity through the use of multiple assays in patients demonstrating an atypical clinical course for anti-GBM disease.

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年12月16日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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年12月15日, ［査読有り］, ［国際誌］
英語

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年11月, ［査読有り］, ［国際誌］
英語

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年07月25日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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年04月17日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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年04月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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.

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年02月04日, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, 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.

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, 2022年01月13日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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, 2021年12月, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, 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.

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年05月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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, 2021年04月, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, 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.

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, 2020年11月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, 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.

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, 2020年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The pleiotropic effects of statins, including an antiarthritic potential, have been noted. This study aimed to determine the efficacy of statins on rheumatoid arthritis (RA) and clarify how statins affect its pathogenesis. Fluvastatin (500 μg/kg/day) or vehicle was given per os to env-pX rats, which carry the human T-cell leukemia virus type I env-pX gene and spontaneously develop destructive arthritis mimicking RA, for 30 days. Blood sampling and ultrasonography (US) of the ankle joints were conducted on days 0, 10, 20, and 30. On day 30, all rats were euthanized, and the ankle joints were subjected to histological analysis. To clarify how fluvastatin affects the pathogenesis of RA, comprehensive serum exosomal microRNA (miRNA) analysis was performed. Gene expression in the primary culture of synovial fibroblasts derived from arthritic rat and human and non-arthritic rat periarticular tissues was determined quantitatively by real-time reverse transcription-polymerase chain reaction (RT-PCR). As a result, the development of arthritis in env-pX rats was significantly suppressed by fluvastatin, which was evident from the viewpoints of serology, US imaging, and histology. Comprehensive serum exosomal miRNA analysis suggested that the expression of Rho GTPase-activating protein 12 (Arhgap12) was decreased in arthritic env-pX rats but increased with the administration of fluvastatin. Corresponding results were obtained by quantitative RT- PCR using primary culture of synovial fibroblasts. The collective findings suggest that fluvastatin prevents the development of arthritis in env-pX rats via the up-regulation of ARHGAP12. This study suggests that ARHGAP12 can be a possible therapeutic target of RA.

Pharmaceutical immunoglobulins reduce neutrophil extracellular trap formation and ameliorate the development of MPO-ANCA-associated vasculitis
Ryo Uozumi, Risa Iguchi, Sakiko Masuda, Yuka Nishibata, Daigo Nakazawa, Utano Tomaru, Akihiro Ishizu
Modern Rheumatology, 30, 3, 544, 550, 2020年05月03日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2019, © 2019 Japan College of Rheumatology. 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.

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, 38, 2, 161, 165, 2020年03月01日, ［査読有り］, ［国際誌］
研究論文（学術雑誌）, 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 and Therapy, 21, 1, 296, 296, 2019年12月21日, ［査読有り］, ［国際誌］
英語

Detection of Increased Vascular Signal in Arthritis-Prone Rats Without Joint Swelling Using Superb Microvascular Imaging Ultrasonography
Tatsunori Horie, Mutsumi Nishida, Shun Tanimura, Tamotsu Kamishima, Erika Tamai, Yutaka Morimura, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
Ultrasound in Medicine and Biology, 45, 8, 2086, 2093, 2019年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2019 World Federation for Ultrasound in Medicine & Biology 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.

Formation and Disordered Degradation of Neutrophil Extracellular Traps in Necrotizing Lesions of Anti-Neutrophil Cytoplasmic Antibody–Associated Vasculitis
Sakiko Masuda, Mayu Nonokawa, Emika Futamata, Yuka Nishibata, Sari Iwasaki, Takahiro Tsuji, Yutaka Hatanaka, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Tamihiro Kawakami, Tatsuya Atsumi, Akihiro Ishizu
American Journal of Pathology, 189, 4, 839, 846, 2019年04月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2019 American Society for Investigative Pathology Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is characterized by the production of ANCAs and systemic necrotizing vasculitis in small vessels. Disordered regulation of neutrophil extracellular traps (NETs) is critically involved in the pathogenesis of AAV. NETs are web-like DNA decorated with antimicrobial proteins; they are extruded from activated neutrophils. The principal degradation factor of NETs in vivo is DNase I; however, NETs resistant to DNase I can persist in tissues and can lead to the production of ANCAs. Deposition of NETs has been demonstrated in glomerular crescents and necrotizing vasculitis in AAV. Here, the amount of NETs in formalin-fixed, paraffin-embedded tissue sections was examined, and the results for AAV were compared with the results for diseases that should be distinguished from AAV. NETs were more abundant in necrotizing vasculitis of AAV than in non–ANCA-associated vasculitis, or in granulomatous angiitis. Pulmonary granulomas in AAV and non–ANCA-associated diseases were further studied. The amount of NETs was significantly greater in necrotizing granulomas of AAV than in granulomas of sarcoidosis without necrosis. Although NETs were formed in necrotizing granulomas of tuberculosis equivalently to those formed in AAV, they were more susceptible to degradation by DNase I than were NETs in AAV. The formation and disordered degradation of NETs in necrotizing lesions are characteristics of AAV and are possibly related to its pathogenesis.

Epitope recognized by anti-glomerular basement membrane (GBM) antibody in a patient with repeated relapse of anti-GBM disease
Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Mandkhai Nergui, Jia Xiao-yu, Zhao Cui, Ming hui Zhao, Kimimasa Nakabayashi, Akihiro Ishizu
Experimental and Molecular Pathology, 107, 165, 170, 2019年04月, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2019 Elsevier Inc. 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.

Detection of Autoreactive Type II NKT Cells: A Pilot Study of Comparison Between Healthy Individuals and Patients with Vasculitis
Yusuke Nishioka, Takaomi Sonoda, Haruki Shida, Yoshihiro Kusunoki, Fumihiko Hattanda, Shun Tanimura, Ryo Uozumi, Mai Yamada, Yuka Nishibata, Sakiko Masuda, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
Cytometry Part A, 93, 11, 1157, 1164, 2018年11月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2018 International Society for Advancement of Cytometry 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.

Brain-derived neurotrophic factor induces angiogenin secretion and nuclear translocation in human umbilical vein endothelial cells
Ayako Mori, Yusuke Nishioka, Mai Yamada, Yuka Nishibata, Sakiko Masuda, Utano Tomaru, Naoyuki Honma, Takanori Moriyama, Akihiro Ishizu
Pathology Research and Practice, 214, 4, 521, 526, 2018年04月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2018 Elsevier GmbH 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.

Vanishing Immunoglobulins: The Formation of Pauci-Immune Lesions in Myeloperoxidase-Antineutrophil Cytoplasmic Antibody-Associated Vasculitis
Emika Futamata, Sakiko Masuda, Yuka Nishibata, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu
Nephron, 138, 4, 328, 330, 2018年03月01日, ［査読有り］, ［国際誌］
英語

Measurement of NET formation in vitro and in vivo by flow cytometry
Sakiko Masuda, Sakika Shimizu, Junji Matsuo, Yuka Nishibata, Yoshihiro Kusunoki, Fumihiko Hattanda, Haruki Shida, Daigo Nakazawa, Utano Tomaru, Tatsuya Atsumi, Akihiro Ishizu
Cytometry Part A, 91, 8, 822, 829, 2017年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © 2017 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC. 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. © 2017 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC.

Establishment of a rat model of thrombosis induced by intravenous injection of antiphosphatidylserine-prothrombin complex antibody
Mai Yamada, Tamihiro Kawakami, Kohei Takashima, Yusuke Nishioka, Yuka Nishibata, Sakiko Masuda, Shigeru Yoshida, Utano Tomaru, Akihiro Ishizu
Rheumatology (United Kingdom), 56, 6, 1013, 1018, 2017年06月01日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, © The Author 2017. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. 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 mg/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.

Measurement of lipoprotein particle sizes using dynamic light scattering
Toshihiro Sakurai, Suchin Trirongjitmoah, Yuka Nishibata, Takeshi Namita, Masahiro Tsuji, Shu Ping Hui, Shigeki Jin, Koichi Shimizu, Hitoshi Chiba
Annals of Clinical Biochemistry, 47, 5, 476, 481, SAGE PUBLICATIONS INC, 2010年09月, ［国際誌］
英語, 研究論文（学術雑誌）, Background: A simple method for the measurement of LDL particle sizes is needed in clinical laboratories because a predominance of small, dense LDL (sd LDL) has been associated with coronary heart disease. We applied dynamic light scattering (DLS) to measure lipoprotein particle sizes, with special reference to sd LDL. Methods: Human serum lipoproteins isolated by a combination of ultracentrifugation and gel chromatography, or by sequential ultracentrifugation, were measured for particle size using DLS. Results: The sizes of polystyrene beads, with diameters of 21 and 28 nm according to the manufacturer, were determined by DLS as 19.3 ± 1.0 nm (mean ± SD, n = 11) and 25.5 ± 1.0 nm, respectively. The coefficients of variation for the 21 and 28 nm beads were 5.1% and 3.8% (within-run, n = 11), and 2.9% and 6.2% (between-run, n = 3), respectively. The lipoprotein sizes determined by DLS for lipoprotein fractions isolated by chromatography were consistent with the elution profile. Whole serum, four isolated lipoprotein fractions (CM + VLDL + IDL, large LDL, sd LDL and HDL) and a non-lipoprotein fraction isolated by sequential ultracentrifugation were determined by DLS to be 13.1 ± 7.5, 37.0 ± 5.2, 21.5 ± 0.8, 20.3 ± 1.1, 8.6 ± 1.5 and 8.8 ± 2.0 nm, respectively. Conclusions: The proposed DLS method can differentiate the sizes of isolated lipoprotein particles, including large LDL and sd LDL, and might be used in clinical laboratories in combination with convenient lipoprotein separation.

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年02月
(一社)日本脈管学会, 日本語

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, 36, Suppl., 192, 192, 2023年05月
(一社)日本小児腎臓病学会, 日本語

ANCA関連血管炎に対するアバコパンを含む新規治療 Cathepsin C阻害による好中球細胞外トラップ形成抑制　　　　　　　　　　　　　　　
西端 友香, 荒井 粋心, 谷口 舞, 益田 紗季子, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 629, 629, 2023年03月
(一社)日本リウマチ学会, 日本語

自己抗体から紐解く疾患の病理病態 ANCA関連血管炎におけるintermolecular epitope spreadingによる抗GBM抗体の産生　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 112, 1, 196, 196, 2023年03月
(一社)日本病理学会, 日本語

ANCA関連血管炎に対するアバコパンを含む新規治療 C5a受容体拮抗薬と好中球エラスターゼ阻害剤の好中球活性化抑制比較　　　　　　　　　　　　　　　
荒井 粋心, 西端 友香, 谷口 舞, 益田 紗季子, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 629, 629, 2023年03月
(一社)日本リウマチ学会, 日本語

ANCA関連血管炎に対するアバコパンを含む新規治療 好中球細胞外トラップにDNase I抵抗性を付与するタンパクの同定　　　　　　　　　　　　　　　
谷口 舞, 益田 紗季子, 中村 哲朗, 荒井 粋心, 西端 友香, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 630, 630, 2023年03月
(一社)日本リウマチ学会, 日本語

ベーチェット病 ベーチェット病における口内炎の発生原因の解明　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 川邊 智宏, 宮前 多佳子, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 649, 649, 2023年03月
(一社)日本リウマチ学会, 日本語

SLEモデルマウスへのステロイドパルスは好中球細胞外トラップを誘導する　　　　　　　　　　　　　　　
小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 886, 886, 2023年03月
(一社)日本リウマチ学会, 日本語

無症候性血尿を呈した抗糸球体基底膜(GBM)抗体陽性症例の血清を用いた抗体解析　　　　　　　　　　　　　　　
西端 友香, 佐藤 雅之, 長森 恒久, 益田 紗季子, 外丸 詩野, 石津 明洋, 脈管学, 63, 1, 13, 14, 2023年02月
(一社)日本脈管学会, 日本語

COVID-19関連を含むIgA血管炎皮膚生検標本を使用したNeutrophil Extracellular Traps(NETs)の検証　　　　　　　　　　　　　　　
川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 石津 明洋, 外丸 詩野, 日本皮膚科学会雑誌, 133, 2, 254, 254, 2023年02月
(公社)日本皮膚科学会, 日本語

感染症と血管炎 COVID-19発症後およびCOVID-19ワクチン接種後IgA血管炎の皮膚生検組織における好中球細胞外トラップの沈着 COVID-19非関連IgA血管炎との比較　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 外丸 詩野, 横山 華英, 池田 高治, 川上 民裕, 石津 明洋, 脈管学, 63, 1, 10, 11, 2023年02月
(一社)日本脈管学会, 日本語

皮膚血管炎動物モデルの完成　　　　　　　　　　　　　　　
川上 民裕, 中出 一生, 田村 宥人, 伊藤 吹夕, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋, 脈管学, 63, 1, 15, 15, 2023年02月
(一社)日本脈管学会, 日本語

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年

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年
(公社)日本皮膚科学会, 日本語

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年12月
(一社)日本皮膚免疫アレルギー学会, 日本語

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年08月
(一社)日本アレルギー学会, 日本語

ベーチェット病皮膚生検標本を使用したNETsの検証　　　　　　　　　　　　　　　
川上 民裕, 横山 華英, 池田 高治, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋, 西日本皮膚科, 84, 4, 371, 371, 2022年08月
日本皮膚科学会-西部支部, 日本語

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年08月
(一社)日本アレルギー学会, 日本語

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, 111, 1, 162, 162, 2022年03月
(一社)日本病理学会, 日本語

好中球細胞外トラップにDNase I抵抗性を付与するタンパクの探索　　　　　　　　　　　　　　　
益田 紗季子, 北野 翔大, 西端 友香, 外丸 詩野, 石津 明洋, 日本病理学会会誌, 111, 1, 214, 214, 2022年03月
(一社)日本病理学会, 日本語

SLE・抗リン脂質抗体症候群:基礎 全身性エリテマトーデスへのステロイドパルスが好中球細胞外トラップ形成に及ぼす影響　　　　　　　　　　　　　　　
小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 347, 347, 2022年03月
(一社)日本リウマチ学会, 日本語

ANCA関連血管炎:基礎研究・予後予測因子 ブルトン型チロシンキナーゼ阻害剤チラブルチニブによるMPO-ANCA関連血管炎誘導モデルの発症抑制　　　　　　　　　　　　　　　
中出 一生, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 406, 406, 2022年03月
(一社)日本リウマチ学会, 日本語

ANCA関連血管炎:基礎研究・予後予測因子 MPO-ANCA関連血管炎モデルにおける新規好中球機能制御化合物薬の抑制効果　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 407, 407, 2022年03月
(一社)日本リウマチ学会, 日本語

SLE・抗リン脂質抗体症候群:基礎 全身性エリテマトーデスへのステロイドパルスが好中球細胞外トラップ形成に及ぼす影響　　　　　　　　　　　　　　　
小川 帆貴, 横田 隼一, 清水 智弘, 西端 友香, 益田 紗季子, 外丸 詩野, 岩崎 倫政, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 66回, 347, 347, 2022年03月
(一社)日本リウマチ学会, 日本語

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年02月
(一社)日本脈管学会, 日本語

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年02月
(一社)日本脈管学会, 日本語

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年01月
(公社)日本皮膚科学会, 日本語

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年01月
(公社)日本皮膚科学会, 日本語

【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年11月
(株)東京医学社, 日本語

【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年11月
(株)文光堂, 日本語

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年10月
(一社)日本臨床免疫学会, 日本語

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年08月
(一社)日本アレルギー学会, 日本語

中小型血管炎:ANCA関連血管炎(基礎) myosin light chain 6を認識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, 65回, 407, 407, 2021年03月
(一社)日本リウマチ学会, 日本語

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年01月
(一社)日本脈管学会, 日本語

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

低酸素環境が好中球細胞外トラップ形成へ与える影響
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年
(一社)日本病理学会, 日本語

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年01月
(一社)日本脈管学会, 日本語

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年09月
(公社)日本整形外科学会, 日本語

中小型血管炎(ANCA関連血管炎) 抗好中球細胞質抗体(ANCA)に続き抗糸球体基底膜(GBM)抗体が産生されるメカニズム　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 64回, 507, 507, 2020年08月
(一社)日本リウマチ学会, 日本語

肺基礎疾患を有する肺炎患者における分解抵抗性好中球細胞外トラップの形成　　　　　　　　　　　　　　　
益田 紗季子, 石橋 美郷, 加藤 くるみ, 西端 友香, 田中 敏, 外丸 詩野, 辻野 一三, 石津 明洋, 日本病理学会会誌, 109, 1, 308, 308, 2020年03月
(一社)日本病理学会, 日本語

抗GBM抗体が認識するエピトープの表出に関する検討　　　　　　　　　　　　　　　
西端 友香, 野々川 茉佑, 益田 紗季子, 中沢 大悟, 田中 敏, 外丸 詩野, 石津 明洋, 脈管学, 60, 2, 19, 19, 2020年02月
(一社)日本脈管学会, 日本語

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年

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年11月
(公社)日本皮膚科学会, 日本語

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年05月
北海道医学会, 日本語

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年04月
(一社)日本病理学会, 日本語

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年04月
(一社)日本病理学会, 日本語

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, 2019年03月
OXFORD UNIV PRESS, 英語, 研究発表ペーパー・要旨（国際会議）

抗糸球体基底膜抗体病(抗GBM病)の頻回再発症例の抗GBMが認識するエピトープ　　　　　　　　　　　　　　　
西端 友香, 東 里緒, 益田 紗季子, 中沢 大悟, 田中 敏, 外丸 詩野, 中林 公正, 石津 明洋, 脈管学, 59, 3, 19, 19, 2019年03月
(一社)日本脈管学会, 日本語

ヒストンは好中球細胞上にLAMP2を表出し、抗LAMP2抗体と連携して皮膚小血管炎の発症機序に関与している　　　　　　　　　　　　　　　
川上 民裕, 竹内 そら, 菊池 彩翔, 西端 友香, 益田 紗季子, 外丸 詩野, 石津 明洋, 脈管学, 59, 3, 22, 22, 2019年03月
(一社)日本脈管学会, 日本語

ANCA関連血管炎の壊死性病変部における好中球細胞外トラップの存在と病的意義　　　　　　　　　　　　　　　
益田 紗季子, 西端 友香, 中沢 大悟, 外丸 詩野, 川上 民裕, 渥美 達也, 石津 明洋, 日本リウマチ学会総会・学術集会プログラム・抄録集, 63回, 1, 777, 777, 2019年03月
(一社)日本リウマチ学会, 日本語

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年

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年

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年11月
北海道医学会, 日本語

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年04月
(一社)日本病理学会, 日本語

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年04月
(一社)日本病理学会, 日本語

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年04月
(一社)日本病理学会, 日本語

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年04月
(一社)日本病理学会, 日本語

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年03月
(一社)日本リウマチ学会, 日本語

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年03月
(一社)日本リウマチ学会, 日本語

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年03月
(一社)日本リウマチ学会, 日本語

ANCA関連血管炎の壊死性病変部におけるNETsの存在と病的意義　　　　　　　　　　　　　　　
益田 紗季子, 野々川 茉佑, 西端 友香, 岩崎 沙理, 辻 隆裕, 田中 敏, 外丸 詩野, 川上 民裕, 石津 明洋, 脈管学, 58, 3, 35, 35, 2018年03月
(一社)日本脈管学会, 日本語

プロテアーゼによる抗糸球体基底膜抗体病の隔絶抗原の表出　　　　　　　　　　　　　　　
西端 友香, 植松 千浩, 益田 紗季子, 田中 敏, 外丸 詩野, 石津 明洋, 脈管学, 58, 3, 36, 37, 2018年03月
(一社)日本脈管学会, 日本語

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年03月
(一社)日本リウマチ学会, 日本語

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年03月
(一社)日本リウマチ学会, 日本語

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年03月
(一社)日本リウマチ学会, 日本語

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年

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年

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年

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年11月
北海道医学会, 日本語

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年03月
OXFORD UNIV PRESS, 英語, 研究発表ペーパー・要旨（国際会議）

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年03月
(一社)日本病理学会, 日本語

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年03月
(一社)日本病理学会, 日本語

血管炎の実験動物モデル 抗PSPT抗体は、正常ラットに血栓を発症させる　　　　　　　　　　　　　　　
川上 民裕, 山田 真衣, 高島 滉平, 西岡 佑介, 西端 友香, 益田 紗季子, 吉田 繁, 外丸 詩野, 石津 明洋, 脈管学, 57, 2, 22, 22, 2017年02月
(一社)日本脈管学会, 日本語

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

自製ELISAによる酸化リポ蛋白の測定とその有用性の検討　　　　　　　　　　　　　　　
櫻井 俊宏, 生田 知子, 古牧 宏啓, 高橋 祐司, 西端 友香, 石川 明彦, 古川 博之, 和田 典男, 永坂 敦, 惠 淑萍, 神 繁樹, 武田 晴治, 布田 博敏, 小林 清一, 千葉 仁志, 臨床病理, 59, 補冊, 204, 204, 2011年10月
(一社)日本臨床検査医学会, 日本語

酸化リポ蛋白自己抗体の研究 臨床的検討
櫻井 俊宏, 西端 友香, 高橋 祐司, 古川 博之, 和田 典夫, 永坂 敦, 惠 淑萍, 生田 知子, 古牧 宏啓, 神 繁樹, 武田 晴治, 布田 博敏, 小林 清一, 千葉 仁志, 臨床病理, 58, 補冊, 94, 94, 2010年07月
(一社)日本臨床検査医学会, 日本語

抗apoE抗体と抗apoCIII抗体のsandwich-ELISAによるレムナントリポ蛋白の検出について
古牧 宏啓, 櫻井 俊宏, 高橋 祐司, 生田 知子, 西端 友香, 惠 淑萍, 神 繁樹, 布田 博敏, 千葉 仁志, 臨床病理, 58, 補冊, 96, 96, 2010年07月
(一社)日本臨床検査医学会, 日本語

酸化リポ蛋白自己抗体の研究 モノクローナル抗体の作製
西端 友香, 櫻井 俊宏, 高橋 祐司, 古川 博之, 和田 典男, 永坂 敦, 惠 淑萍, 生田 知子, 古牧 宏啓, 神 繁樹, 武田 晴治, 布田 博敏, 小林 清一, 千葉 仁志, 臨床病理, 58, 補冊, 173, 173, 2010年07月
(一社)日本臨床検査医学会, 日本語

酸化リポ蛋白の新規測定法の開発　　　　　　　　　　　　　　　
櫻井 俊宏, 生田 知子, 古牧 宏啓, 高橋 祐司, 西端 友香, 古川 博之, 和田 典男, 永坂 敦, 惠 淑萍, 神 繁樹, 武田 晴治, 布田 博敏, 小林 清一, 千葉 仁志, 臨床化学, 39, Suppl.1, 121, 121, 2010年07月
(一社)日本臨床化学会, 日本語

apoCIIIとapoEの両者を持つリポ蛋白粒子(Lp-CIII-E)の分布の検討　　　　　　　　　　　　　　　
古牧 宏啓, 櫻井 俊宏, 高橋 祐司, 生田 知子, 西端 友香, 惠 淑萍, 神 繁樹, 布田 博敏, 武田 晴治, 千葉 仁志, 臨床化学, 39, Suppl.1, 123, 123, 2010年07月
(一社)日本臨床化学会, 日本語

動的光散乱法を用いた各種リポ蛋白粒径の計測
西端 友香, 櫻井 俊宏, Suchin Trirongjitmoah, 浪田 健, 神 繁樹, 清水 孝一, 千葉 仁志, 臨床病理, 57, 補冊, 241, 241, 2009年07月
(一社)日本臨床検査医学会, 日本語

動的光散乱法を用いたリポ蛋白粒径計測法　　　　　　　　　　　　　　　
西端 友香, 櫻井 俊宏, Suchin Trirongjitmoah, 浪田 健, 神 繁樹, 清水 孝一, 千葉 仁志, 臨床化学, 38, Suppl.1, 149, 149, 2009年07月
(一社)日本臨床化学会, 日本語

原子間力顕微鏡を用いたリポ蛋白粒径計測法　　　　　　　　　　　　　　　
櫻井 俊宏, Suchin Trirongjitmoah, 西端 友香, 神 繁樹, 清水 孝一, 千葉 仁志, 臨床化学, 38, Suppl.1, 150, 150, 2009年07月
(一社)日本臨床化学会, 日本語

動的光散乱によるLDLの計測 sdLDLの定量評価をめざして(Application of Dynamic Light Scattering to LDL Measurement: Attempt for quantitative evaluation of small dense LDL)
トライロンジットモア・スチン, 櫻井 俊宏, 西端 友香, 浪田 健, 加藤 祐次, 飯永 一也, 千葉 仁志, 清水 孝一, 電子情報通信学会技術研究報告(MEとバイオサイバネティックス), 109, 106, 23, 28, 2009年06月
LDL(low density lipoprotein)の中でも、小型で比重の大きいsdLDL(small dense LDL)は、心疾患との高い相関から、臨床検査における重要性が指摘されている。従来法は煩雑で長時間を要する。このsdLDLの定量評価をめざし、動的光散乱(DLS:dynamic light scattering)の応用を考えた。DLS測定により得られる平均粒径や相関関数を用いてLDL中のsdLDL量の割合を求める手法を3種新たに考案した。sdLDLの割合を直接求める数式解を導出した。粒径の異なるラテックス標準粒子および実際のLDLを用いた実測により、提案手法の有効性を実証した。(著者抄録), (一社)電子情報通信学会, 英語

動的光散乱によるsmall dense LDLの割合推定の試み　　　　　　　　　　　　　　　
トライロンジットモア・スチン, 浪田 健, 西端 友香, 櫻井 俊宏, 加藤 祐次, 飯永 一也, 千葉 仁志, 清水 孝一, 生体医工学, 47, 3, 321, 321, 2009年06月
(公社)日本生体医工学会, 日本語

動的光散乱を用いた各種リポ蛋白粒径の簡易計測法の開発　　　　　　　　　　　　　　　
櫻井 俊宏, 西端 友香, Trirongjitmoah Suchin, 浪田 健, 清水 孝一, 千葉 仁志, 臨床化学, 37, Suppl.1, 105, 105, 2008年08月
(一社)日本臨床化学会, 日本語

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, 2023年03月, (一社)日本リウマチ学会, 日本語
2023年03月 - 2023年03月

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, 2023年03月, (一社)日本病理学会, 日本語
2023年03月 - 2023年03月

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, 2023年02月, (一社)日本脈管学会, 日本語
2023年02月 - 2023年02月

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, 2022年03月, (一社)日本リウマチ学会, 日本語
2022年03月 - 2022年03月

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, 2021年
2021年 - 2021年

中小型血管炎(ANCA関連血管炎) 抗好中球細胞質抗体(ANCA)に続き抗糸球体基底膜(GBM)抗体が産生されるメカニズム　　　　　　　　　　　　　　　
西端 友香, 益田 紗季子, 中沢 大悟, 外丸 詩野, 石津 明洋
日本リウマチ学会総会・学術集会プログラム・抄録集, 2020年08月, (一社)日本リウマチ学会, 日本語
2020年08月 - 2020年08月

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, 2020年02月, (一社)日本脈管学会, 日本語
2020年02月 - 2020年02月

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, 2019年04月, (一社)日本病理学会, 日本語
2019年04月 - 2019年04月

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, 2019年03月, (一社)日本脈管学会, 日本語
2019年03月 - 2019年03月

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, 2018年03月, (一社)日本脈管学会, 日本語
2018年03月 - 2018年03月

動的光散乱法を用いたリポ蛋白粒径計測法　　　　　　　　　　　　　　　
西端 友香, 櫻井 俊宏, Suchin Trirongjitmoah, 浪田 健, 神 繁樹, 清水 孝一, 千葉 仁志
臨床化学, 2009年07月, (一社)日本臨床化学会, 日本語
2009年07月 - 2009年07月

動的光散乱法を用いた各種リポ蛋白粒径の計測
西端 友香, 櫻井 俊宏, Suchin Trirongjitmoah, 浪田 健, 神 繁樹, 清水 孝一, 千葉 仁志
臨床病理, 2009年07月, (一社)日本臨床検査医学会, 日本語
2009年07月 - 2009年07月

現在
日本病理学会　　　　　　　　　　　　　　　

ホスファチジルセリン依存性抗プロトロンビン抗体の好中球細胞外トラップ誘導能の解明
科学研究費助成事業
2024年04月01日 - 2027年03月31日
西端 友香
日本学術振興会, 若手研究, 北海道大学, 24K19231

学士課程, 医学部（保健学科）

修士課程, 保健科学院

博士課程, 保健科学院