抗リン脂質抗体症候群:病因の解明と新たなる治療ターゲット

2007 Fiscal Year Annual Research Report

• Project/Area Number: 19390269
• Research Institution: Hokkaido University
• Principal Investigator: 小池 隆夫 Hokkaido University, 大学院・医学研究科, 教授 (80146795)
• Keywords: 抗リン脂質抗体症候群 / β_2GPI / 血栓形成 / プロテオーム解析 / シグナル伝達 / p38MAPK / ゲルゾリン / インテグリン

Research Abstract

抗リン脂質抗体による細胞刺激シグナル、細胞活性化メカニズムについて検討を行うことにより抗リン脂質抗体症候群発症機構を解明し、さらにシグナル伝達制御による病態制御を可能とすることを目的とした。β_2GPIと結合する細胞表面タンパク質の検出のため、リコンビナントFLAGタグβ2GPIを、RAW264.7の細胞溶解液と混合し、抗FLAG抗体アフィニティークロマトグラフィーによりFLAGタグβ_2GPIと複合体を形成するタンパク質(β_2GPI関連プロテオーム)を網羅的に分離した。MALDI-TOFもしくはESI-LC-MS/MSにより解析し、結果として得られたタンパク質につき、β_2GPIとの結合をFACS法およびウエスタンブロッティング法にて確認をした。さらにそのタンパク質の存在の有無による抗CL/β_2GPI抗体による細胞刺激シグナルの変動について、インテグリン直下のFAKのリン酸化をウエスタンブロッティングにて解析した。次に、RAW264.7にNF一κBレポーターベクターをトランスフェクションし、ルシフェラーゼアッセイにて解析した。
その結果、ゲルゾリンがβ_2GPIと結合していることがウエスタンブロッティング法にて確認された。また、フローサイトメーターによりゲルゾリンが細胞膜表面上でインテグリンα5β1と結合することが確認され、さらにβ_2GPIがゲルゾリン、インテグリンα5β1を含む複合体と結合することが確認された。また、抗β_2GPI抗体刺激によりインテグリン直下のFAKのリン酸化の亢進が認められ,抗インテグリン抗体の存在下ではFAKのリン酸化が減弱した。ルシフェラーゼアッセイにより、抗インテグリン抗体によるインテグリンα5β1阻害により、抗CL/β_2GPI抗体による細胞刺激シグナルが減弱することが確認された。

Research Products

• [Journal Article] Reversible cardiomyopathy due to secondary hemochromatosis with multitransfusions for severe aplastic anemia after successful non-myeloablative stem cell transplantation. (2008)
  • Author(s): Nishio M, Endo T, Nakao S, Sato N, Koike T.
  • Journal Title: Int J Cardiol (in press)
  • Peer Reviewed
• [Journal Article] Polymyalgia rheumatica as the manifestation of unclassified aortits. (2007)
  • Author(s): Kataoka H, Atsumi T, Hashimoto T, Horita T, Yasuda S, Koike T.
  • Journal Title: Mod Rheumatol 18(1) Pages: 105-108
  • Peer Reviewed
• [Journal Article] Multiple autoimmune diseases after autologous stem-cell transplantation. (2007)
  • Author(s): Bohgaki T, Atsumi T, Koike T.
  • Journal Title: New Engl J Med. 357(26) Pages: 2734-2736
  • Peer Reviewed
• [Journal Article] Melanocortin 2 receptor is required for adrenal gland development, steroidogenesis, and neonatal gluconeogenesis. (2007)
  • Author(s): Chida D, Nakagawa S, Nagai S, Sagara H, Katsumata H, Imaki T, Suzuki H, Mitani F, Ogishima T, Shimizu C, Kotaki H, Kakuta S, Sudo K, Koike T, Kubo M, Iwakura Y.
  • Journal Title: PNAS. 104(46) Pages: 18205-18210
  • Peer Reviewed
• [Journal Article] Successful efavirenz dose reduction in HIV type 1-infected individuals with cytochrome P450 2B6 *6 and *26. (2007)
  • Author(s): Gatanaga H, Hayashida T, Tsuchiya K, Yoshino M, Kuwahara T, Tsukada H, Fujimoto K, Sato I, Ueda M, Horiba M, Hamaguchi M, Yamamoto M, Takata N, Kimura A, Koike T, Gejyo F, Matsushita S, Shrasaka T, Kimura S, Oka S.
  • Journal Title: CID 45(9) Pages: 1230-1237
  • Peer Reviewed
• [Journal Article] Soluble fibrin monomer degradation products as a potentially useful markerfor hypercoagulable states with accelerated fibrinolysis. (2007)
  • Author(s): eko M, Nakabayashi T, Tarumi T, Naito S, Yoshida M, Kanazawa K, Mizukami K, Koike T.
  • Journal Title: Clin Chim Acta. 386(1-2) Pages: 38-45
  • Peer Reviewed
• [Journal Article] Defective expression of ras guanyl nucleotide-releasing protein 1 in a subset of patients with systemic lupus erythematosus. (2007)
  • Author(s): Yasuda S, Stevens RL, Terada T, Takeda M, Hashimoto T, Fukae J, Horita T, Kataoka H, Atsumi T, Koike T.
  • Journal Title: J. Immunol 179(7) Pages: 4890-4900
  • Peer Reviewed
• [Journal Article] The Proinflammatory cytokine macrophage migration inhibitory factor regulates glucose metabolism during systemic inflammation. (2007)
  • Author(s): Atsumi T, Cho YR, Leng L, McDonald C, Yu T, Danton C, Hong EG, Mitchell RA, Metz C, Niwa H, Takeuchi J, Onodera S, Umino T, Yoshioka N, Koike T, Kim JK, Bucala R.
  • Journal Title: J. Immunol 179(8) Pages: 5399-5406,
  • Peer Reviewed
• [Journal Article] Increased fructose 2, 6-bisphosphate in peripheral blood mononuclear cells of patients with diabetes. (2007)
  • Author(s): Atsumi T, Chiba H, Yoshioka N, Bucala R, Koike T.
  • Journal Title: Endocr J. 57(4) Pages: 517-520
  • Peer Reviewed
• [Journal Article] Amicrobial pustulosis associated with IgA nephropathy and Sjogren's syndrome. (2007)
  • Author(s): Natsuga K, Sawamura D, Homma E, Nomura T, Abe M, Muramatsu R, Mochizuki T, Koike T, Shimizu H.
  • Journal Title: J Am Acad Dermatol 57 Pages: 523-526
  • Peer Reviewed
• [Journal Article] Human monoclonal antibodies against the complex of phosphatidylserine and prothrombin from patients with the antiphospholipid antibodies. (2007)
  • Author(s): Horita T, Ichikawa K, Kataoka H, Yasuda S, Atsumi T, Koike T.
  • Journal Title: Lupus. 16(7) Pages: 509-516
  • Peer Reviewed
• [Journal Article] A polymorphism in human platelet antigen 6b and risk of thrombocytopenia in patients with systemic lupus erythematosus. (2007)
  • Author(s): Amengual O, Atsumi T, Komano Y, Kataoka H, Horita T, Yasuda S, KoikeT.
  • Journal Title: Arthritis Rheum. 56(8) Pages: 2803-2809
  • Peer Reviewed
• [Journal Article] Acute onset of type 1 diabetes accompanied by acute hepatitis C: the potential role of proinflammatory cytokine in the pathogenesis of autoimmune diabetes. (2007)
  • Author(s): Masuda H, Atsumi T, Fujisaku A, Shimizu C, Yoshioka N, Koike T.
  • Journal Title: Diabetes Res Clin Pract 75(3) Pages: 357-361
  • Peer Reviewed
• [Journal Article] Delayed redistribution of CD27, CD40 and CD80 positive B cells and the impaired in vitro immunoglobulin production in patients with non-Hodgkinlymphoma after rituximab treatment as an adjuvant to autologous stem cell transplantation. (2007)
  • Author(s): Nishio M, Fujimoto K, Yamamoto S, Endo T, Sakai T, Obara M, Kumano K, Yamaguchi K, Takeda Y, Goto H, Sato N, Koizumi K, Mukai M, Koike T.
  • Journal Title: Br J Haematol 137(4) Pages: 349-354
  • Peer Reviewed
• [Journal Article] Hepatosplenic alpha/beta T cell lymphoma presenting with cold agglutinin disease. (2007)
  • Author(s): Minauchi K, Nishio M, Itoh T, Yamamoto S, Fujimoto K, Sato N, Koike T.
  • Journal Title: Ann Hematol. 86(2) Pages: 155-157
  • Peer Reviewed
• [Journal Article] Unilateral adrenalectomy improves insulin resistance and polycystic ovaries in a middle-aged woman with virilizing adrenocortical adenoma complicated with Cushing's syndrome. (2007)
  • Author(s): Nakamura A, Shimizu C, Nagai S, Taniguchi S, Umetsu M, Atsumi T, Wada N, Yoshioka N, Ono Y, Sasano H, Koike T.
  • Journal Title: J Endocrinol Invest 30 Pages: 65-69
  • Peer Reviewed
• [Journal Article] "Resurrection of thrombin" in the pathophysiology of the antiphospholipid syndrome. (2007)
  • Author(s): Koike T, Atsumi T.
  • Journal Title: Arthritis Rheum 56(2) Pages: 393-394
  • Peer Reviewed
• [Presentation] Multiple autoimmune diseases after CD34+ selected autologous hematopoietic stem cell transplantation in a patient with systemic sclerosis (2008)
  • Author(s): Koike, T. :
  • Organizer: The Mosaic of Autoimmunity
  • Place of Presentation: Kfar Maccabiah Olympic Village, Israel,
  • Year and Date: 2008-02-11
• [Presentation] Recent Progress in Systemic Autommune Diseases (2007)
  • Author(s): Koike, T
  • Organizer: 第8回国際SLE学会
  • Place of Presentation: Shanghai international Convention Center, 上海
  • Year and Date: 20070523-25
• [Presentation] Antiphospholipid antibodies and cell acyivation (2007)
  • Author(s): Koike, T.
  • Organizer: 12th International Congress on Antiphospholipid Antibodies
  • Place of Presentation: Palazzo dei Congressi, Florence, Italy
  • Year and Date: 2007-04-18