Curriculum Vitaes

Keiji Miura

  (三浦 惠二)

Profile Information

Affiliation
School of Health Sciences Faculty of Rehabilitation, Fujita Health University
Degree
理学博士(名古屋大学)

Researcher number
20199946
J-GLOBAL ID
201101052461367514
researchmap Member ID
B000004116

External link

自己免疫疾患を対象に研究を行っています。
自己免疫疾患患者の血清を調べると、血管内皮細胞の細胞表面に結合する抗体(抗血管内皮細胞抗体;AECA)が高頻度で見つかってきます。細胞への抗体の結合が病態に直接関係してくると考えられ、結合している分子すなわち自己抗原を特定するための研究が行われてきました。徐々に見つかってきてはいるものの、まだまだ未知の自己抗原が多いと考えられます。それらの分子を特定し、病態との関連を解明すること、そして新たな診断法開発を目標にしています。
その基盤技術となる方法として、CSP-ELISA法を独自に開発し、特許を取得しています。(特開2014-081365)この方法により、立体構造が不安定になりやすい膜タンパク質と抗体との結合を検出することが可能になりました。膜タンパク質に対する抗体取得は、抗がん剤としての利用に繋がる可能性もあり、がん研究の分野での発展も目指しています。

教育では、保健衛生学部1年生を対象に生命科学と化学の講義を担当しています。生物学と化学は医療系専門科目につながる重要な基礎科学ですが、受験に生物や化学を選択しなかった学生もいるため、生物学や化学の基礎を確認しつつ、生命の不思議・おもしろさ・身の回りで起こる自然現象を話題に挙げ、生物であるヒトすなわち自分を見つめる機会にして欲しいと思っています。


Papers

 27
  • Sumitomo-Kondo M, Ukai Y, Iba Y, Ohshima N, Miura K, Takasaki A, Kurosawa Y, Kurosawa G
    Biochemical and biophysical research communications, 503(2) 1141-1147, Jun, 2018  Peer-reviewed
  • Ayako Kondo, Kazuo Takahashi, Tomohiro Mizuno, Akihiro Kato, Daisuke Hirano, Naoki Yamamoto, Hiroki Hayashi, Shigehisa Koide, Hiroshi Takahashi, Midori Hasegawa, Yoshiyuki Hiki, Shunji Yoshida, Keiji Miura, Yukio Yuzawa
    PLoS One, 11(10) e0163085, Oct, 2016  Peer-reviewed
    Anti-endothelial cell antibodies (AECA) are frequently detected in patients with systemic lupus erythematosus (SLE), but their pathological role remains unclear. We recently developed a solubilized cell surface protein capture enzyme-linked immunosorbent assay (CSPELISA) to detect antibodies against membrane proteins involved in autoimmune reactions. In this study, sera from 51 patients with biopsy-proven lupus nephritis (LN), 25 with SLE without renal involvement (non-LN SLE), 42 disease control (DC) subjects, and 80 healthy control (HC) subjects were tested for IgG-and IgA-AECA for human umbilical vein endothelial cells (HUVEC) and human glomerular EC (HGEC) by using CSP-ELISA. IgG-and IgA-AECA titers were significantly higher in LN and non-LN SLE patients than in the DC or HC (P < 0.001) groups. IgG-and IgA-AECA titers for HUVEC corresponded well with those for HGEC. The IgA-AECA level correlated with the SLE disease activity index and with histological evidence of active lesions (cellular proliferations, hyaline thrombi and wire loops, leukocytic infiltration, and fibrinoid necrosis) in LN patients (P < 0.001). The sensitivity of IgA-AECA as a diagnostic test for histological evidence of active lesions in LN patients was 0.92, with a specificity of 0.70. The significant correlation of IgA-AECA with glomerular hypercellularity indicates that IgA-AECA are associated with endothelial damage in LN.
  • 近藤 亜矢子, 高橋 和男, 秋山 真一, 林 宏樹, 小出 滋久, 富田 亮, 長谷川 みどり, 比企 能之, 三浦 惠二, 湯澤 由紀夫
    日本腎臓学会誌, 55(3) 332-332, Apr, 2013  Peer-reviewed
  • Keiji Miura, Keiko Aoun, Shunji Yoshida, Yoshikazu Kurosawa
    JOURNAL OF IMMUNOLOGICAL METHODS, 382(1-2) 32-39, Aug, 2012  Peer-reviewed
    This article describes a novel method for detecting anti-endothelial cell antibodies (AECAs). Sera from patients with systemic vasculitis or inflammatory conditions have been reported to contain antibodies (Abs) that bind to endothelial cells (EC), i.e., AECAs. AECAs are known to play immunogenic effects by triggering EC activation and vascular damage, but the immunopathological role of AECAs is not clear. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting have previously been used for detecting target antigens of AECAs. However, we assumed that these methods are not appropriate for searching genuine target antigens (Ags) on cell surface, and developed a novel solubilized cell surface protein-capture ELISA (CSP-ELISA). Ags were obtained as cell surface proteins from the plasma membrane of human umbilical vein endothelial cells (HUVECs); these cell surface proteins were biotinylated, solubilized with detergent, and captured on ELISA wells coated with NeutrAvidin (TM) biotin binding protein (NeuAvi). AECA titers in serum from 126 autoimmune disease patients and 122 healthy donors were tested. AECAs were detected in 28 of 36 (78%) of systemic lupus erythematosus (SLE) patients; in 13 of 16(81%) of mixed connective tissue disease (MCTD) patients; and in 5 of 9 (56%) of systemic sclerosis (SSc) patients. Relatively weak denaturation of antigens on ELISA wells caused loss of binding of these autoantibodies (autoAbs). Thus, this newly developed CSP-ELISA method enables the detection of Abs to the labile epitopes of autoantigens (autoAgs) such as membrane proteins, and this method is generally applicable to various kinds of membrane proteins and the Abs against them. We propose CSP-ELISA for measuring AECAs in serum samples for routine laboratory testing. (C) 2012 Elsevier B.V. All rights reserved.
  • Gene Kurosawa, Naofumi Takamatsu, Masayoshi Takahashi, Mariko Sumitomo, Emi Sanaka, Kouji Yamada, Kazuhiro Nishii, Masaru Matsuda, Shuichi Asakawa, Hiroshi Ishiguro, Keiji Miura, Yoshikazu Kurosawa, Nobuyoshi Shimizu, Yuji Kohara, Hiroshi Hori
    GENE, 376(2) 298-299, Jul, 2006  Peer-reviewed

Misc.

 13

Presentations

 6

Teaching Experience

 4

Research Projects

 9

Industrial Property Rights

 7

Other

 1
  • 細胞表面タンパクを抗原とする抗体を測定する方法(CSP-ELISA法) 自己抗体の中には、細胞表面の膜タンパクに結合するものがあり、病態を理解するためには標的抗原を同定して解析する必要がある。膜タンパクの抗原性を保持した形で抗原抗体反応を検出する方法がCSP-ELISA法であり、それを利用することで疾患の新たなバイオマーカーの同定が可能になる。国内特許取得済(特許6327662) *本研究シーズに関する産学共同研究の問い合わせは、藤田医科大学産学連携推進センター(fuji-san@fujita-hu.ac.jp)まで