シェーグレン症候群発症の分子機構:M3Rに対する免疫応答解析

2008 Fiscal Year Annual Research Report

• Project/Area Number: 19591151
• Research Institution: University of Tsukuba
• Principal Investigator: 住田 孝之 University of Tsukuba, 大学院・人間総合科学研究科, 教授 (00183054)
• Keywords: シェーグレン症候群 / M3R / 抗M3R抗体 / B細胞エピトープ / 唾液分泌 / 唾液腺炎モデルマウス / T細胞エピトープ / アナログペプチド

Research Abstract

本研究では、シェーグレン症候群(SS)患者において、ムスカリン作働性アセチルコリン受容体M3(M3R)に対する免疫応答を解析することにより、SSにおける唾液腺破壊の発症機序を解明し、SSに対する新しい抗原特異的治療戦略を開発することを目的とした。本年度は以下の事実を明らかにしてきた。
1)抗M3R抗体の対応B細胞エピトープ解析:SS患者血清を用いて、M3RのN領域、細胞外第一、第二、第三ドメインに対する抗M3R抗体をELISA法にて解析した。その結果、それぞれのドメインに対する自己抗体が存在すること、同一患者において複数のB細胞エピトープがM3R分子上に存在すること、を明らかにした。
2)M3R上のB細胞エピトープと機能:唾液腺上皮細胞由来細胞株(HSG)を塩酸セビメリンで刺激し細胞内のCa2+増減をフォローサイトメトリーで測定する系を確立し、抗M3R抗体の唾液分泌に対する機能について検討した。その結果、M3R第ニドメインに対する抗体はCa2+influxを抑制し、N領域、第一ドメインに対する抗体は増強、第三ドメインに対する抗体は影響を与えない事が判明した。すなわち、対応B細胞エピトープにより自己抗体の機能が異なる事が明らかとなった。
3)SSのモデルマウスの樹立:C57BL/6マウスバックのM3RノックアウトマウスにM3Rの各領域(N、細胞外第一、第二、第三)をコードする合成アミノ酸を混合免疫し、20日目に脾臓細胞をRag1ノックアウトマウスに細胞移入した。30日目以降に単核球浸潤を中心とした唾液腺炎が認められた。浸潤細胞の主体はCD4+T細胞であり、SSの唾液腺炎に類似した組織像を呈していた。以上のように、2つのノックアウトマウスを組み合わせる事により、M3R免疫唾液腺炎モデルマウスを確立することに成功した。

Research Products

• [Journal Article] B cells play a crucial role as antigen-presenting cells and collaborate with in flammatory cytokines in glucose-6-phosphate isomerase-induced arthritis. (2009)
  • Author(s): Tanaka-Watanabe Y, Matsumoto I, Iwanami K, Inoue A, Goto D, Ito S, Tsutsumi A, Sumida T.
  • Journal Title: Clin. Exp. Immunol. 155 Pages: 285-294
  • Peer Reviewed
• [Journal Article] Replication of the association between the C8orf13-BLK region and systemic lupus erythematosus in a Japanese population. (2009)
  • Author(s): Ito I, Kawasaki A, Ito S. Hayashi T, Goto D, Matsumoto I, Tsutsumi A, Hom G, Graham RR, Takasaki Y, Hashimoto H, Ohashi J, Behrens TW, Sumida T, Tsuchiya N.
  • Journal Title: Arthritis Rheum. 60 Pages: 553-558
  • Peer Reviewed
• [Journal Article] Arthritogenic T cell epitope in glucose-6-phosphate isomerase (GPI)-induced arthritis. (2008)
  • Author(s): Iwanami K, Matsum I, Tanaka Y, Inoue A, Goto D, Ito S, Tsutsumi A, Sumida T.
  • Journal Title: Arthritis Res. Ther. 10(Epub) Pages: R130
  • Peer Reviewed
• [Journal Article] Role of STAT4 polymorphisms in systemic lupus erytheMatosus in a Japanese population: a case-control association study of STAT1-STAT4 region. (2008)
  • Author(s): Kawasaki A, Ito I, Hikami K, Ohashi J, Hayashi T,Goto D, Matsumoto I, Ito S, Tsutsumi A, Koga M, Arinami T, Graham R R, Hom G, Takasaki Y, Hashimoto H, Behrens T W, Sumida T and Tsuchiya N.
  • Journal Title: Arthritis Res Ther 10(Epub) Pages: R113
  • Peer Reviewed
• [Journal Article] Muscarinic-3 acetylcholine receptor autoantibody in patients with systemic sclerosis: contribution to severe gastrointestinal tract dysmotility. (2008)
  • Author(s): Kawaguchi Y, Wakamatsu E, Matsumoto I, Nishimagi E, Kamatani N, Satoh T, Kuwana M, Sumida T, Hara M.
  • Journal Title: Ann. Rheum. Dis. Sep1(Epub)
  • Peer Reviewed
• [Journal Article] Therapeutic effects of antibodies to: TNF alpha and IL-6 and CTLA-4 Ig in mice with glucose-6-phosphate isomerase-induced arthritis. (2008)
  • Author(s): Matsumoto I, Zhang H, Yasukochi I, wanami K, Tanaka Y, Inoue A, Goto D, Ito S, Tsutsumi A, Sumida T.
  • Journal Title: Arthritis Res Ther 10(Epub) Pages: R66
  • Peer Reviewed
• [Journal Article] Invariant NKT cells produce IL-17 through IL-23-dependent and-independent pathways with potential modulation of Th17 response in collagen-induced arthritis. (2008)
  • Author(s): Yoshiga Y, Goto D, Segawa S, Ohnishi Y, I. Matsumoto I, Ito S, Tsutsumi A, Taniguchi M and SumidaT.
  • Journal Title: Int. J. Mol. Med. 22 Pages: 369-374
  • Peer Reviewed
• [Journal Article] Crucial role of IL-6/IL-17 cytokine axis in the induction of arthritis by glucose-6-phosphate-isomerase. (2008)
  • Author(s): Iwanami K, Matsumoto I, Watanabe Y, Mihara M, Ohsugi Y, Mamura M, Goto D, Ito S, Tsutsumi A, Kishimoto T, SumTida T.
  • Journal Title: Arthritis Rheum. 58 Pages: 754-763
  • Peer Reviewed
• [Journal Article] Visfatin (pre-B cell colony-enhancing factor) gene expressionin patients with rheumatoid arthritis. (2008)
  • Author(s): Matsui H, Tsutsumi A, Sugihara M, Suzuki T, Iwanami K, Kohno M, Goto D, Matsumoto I, Ito S, Sumida T.
  • Journal Title: Ann. Rheum. Dis. 67 Pages: 571-572
  • Peer Reviewed
• [Journal Article] High prevalence of autoantibodies to muscarinic-3 acetylcholine receptor in patients with juvenile-onset Sjogren's syndrome (2008)
  • Author(s): Nakamura Y. Wakamatsu E, Matsumoto I, Tomiita M, Kohno Y, Mori M, Yokota S, Goto D, Ito S, Tsutsumi A, Sumida T.
  • Journal Title: Ann. Rheum. Dis. 67 Pages: 136-137
  • Peer Reviewed
• [Presentation] The Expression of MicroRNA-424 in Rheumatoid Arthritis. (2008)
  • Author(s): Yoshitaka T, Nakasa T, Inoue A, Okubo A, Takemoto F, Miyaki S, Nishida K, Ochi M, Ozaki T, Sumida T, Asahara H.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-28
• [Presentation] The Pathogenesis of Anti-3 Muscarinic Acetylcholine Receptor Antibodies in Patients with Sjogren's Syndrome. (2008)
  • Author(s): Tsuboi H, Matsumoto I, Wakamatsu E, Iizuka M, Nakamura Y, Hayashi T, Goto D, Ito S, Sumida T.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-28
• [Presentation] Establishment Of New Murine Model For Sjogren'S Syndrome Using M3 Muscarinic Acethylcholine Receptor Knockout Mice. (2008)
  • Author(s): Wakamatsu E, Matsumoto I, Iizuka M, Tsuboi H, Nakamura Y, Hayashi T, Goto D, Ito S, Sumida T.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-28
• [Presentation] Overexpression Of TNFalpha-induced-adipose Related Protein (TIARP) In Joints With Autoimmune Arthritis Mediated By Glucose-6-phophate Isomerase. (2008)
  • Author(s): Inoue A, Matsutomo, I, Iwanami K, Tanaka-Watanabe Y, Hayashi T, Goto D, Ito S, Sumida T.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-27
• [Presentation] Replication of the Association between C8orf13-BLK Region and Systemic Lupus Erythematosus in a Japanese Population. (2008)
  • Author(s): Ito I, Kawasaki A Hayashi T, Matsumoto I, Ito S, Tsutsumi A, Ohashi J, Hom G, Graham R R, Takasaki Y, Hashimoto H, Behrens TW, Sumida T, Tsuchiya N.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-26
• [Presentation] Association of STAT4 polymorphisms with Systemic Lupus Erythematosus in a Japanese Population. (2008)
  • Author(s): Kawasaki A, Ito I, Hikami K, Hayashi T, Matsumoto I, Ito S, Tsutsumi A, Koga M, Arinami T, Graham R R, Hom G, Ohashi J, Takasaki Y, Hashimoto H, Behrens TW, Sumida T, Tsuchiya N.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-26
• [Presentation] Use of Laser Microdissection in the Analysis of Renal-infiltratsing T cell in lupus nephritis. (2008)
  • Author(s): Wang Y, Ito S, Chino Y, Goto D, Matsumoto I, Hayashi T, Sumida T.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-26
• [Presentation] Muscarinic-3 Acetylcholine Receptor Autoantibody in Patients with Systemic Sclerosis. (2008)
  • Author(s): Kawaguchi Y, Nakamura Y, Matsumoto I, Ota Y, Nishimagi E, Kamatani N, Satoh T, Kuwana M, Sumida T, Hara M.
  • Organizer: 2008 ANNUAL SCIENTIFIC MEETING on the ACR
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-26
• [Presentation] Induction of experimental autoimmune sialoadenitis using organ-specific autoantigen knockout mice. (2008)
  • Author(s): Wakamatsu E, Matsumoto I, Iizuka M, Tsuboi H, Nakamura Y, Hayashi T, Goto D, Ito S, Sumida T.
  • Organizer: APLAR2008
  • Place of Presentation: Yokohama
  • Year and Date: 2008-09-26
• [Presentation] Evaluation of anti-TNF biologiics on rheumatoid arthritis using the new low-field extremity MRI and scoring system.APLAR2008 (2008)
  • Author(s): Suzuki T, Hand S, Kose K, Hayashi T, Goto D, Matsumoto I, Ito S, Sumida T.
  • Organizer: APLAR2008
  • Place of Presentation: Yokohama
  • Year and Date: 2008-09-26
• [Presentation] TNF alpha induces human six-transmembrane epitherial antigen of prostate 4 in rheumatoid synovium. (2008)
  • Author(s): Tanaka-Watanabe Y, Matsumoto I, Iwanami K, Inoue A, Minami R, Hayashi T, Goto. D, Ito S, Sumida T.
  • Organizer: APLAR2008
  • Place of Presentation: Yokohama
  • Year and Date: 2008-09-24
• [Book] やさしいシェーグレン症候群の自己管理 (2008)
  • Author(s): 住田孝之
  • Total Pages: 10-14
  • Publisher: 医薬ジャーナル社