T細胞の抗原認識と活性化制御

• Project/Area Number: 12051203
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Chiba University
• Principal Investigator: 斉藤 隆 千葉大学, 大学院・医学研究院, 教授 (50205655)
• Co-Investigator(Kenkyū-buntansha): 西村 泰治 熊本大学, 大学院・医学研究科, 教授 (10156119) ; 湊 長博 京都大学, 大学院・生命科学研究科, 教授 (40137716) ; 中山 俊憲 千葉大学, 大学院・医学研究院, 教授 (50237468)
• Project Period (FY): 2000 – 2002
• Project Status: Completed (Fiscal Year 2002)
• Budget Amount: ¥193,400,000 (Direct Cost: ¥193,400,000); Fiscal Year 2002: ¥66,900,000 (Direct Cost: ¥66,900,000); Fiscal Year 2001: ¥68,400,000 (Direct Cost: ¥68,400,000); Fiscal Year 2000: ¥58,100,000 (Direct Cost: ¥58,100,000)
• Keywords: 部分アゴニストペプチド / T細胞エピトープ / 免疫シナプス形成 / 制御性アダプター分子 / インテグリン活性化 / 慢性骨髄性白血 / Th2分化 / ポリコーム遺伝子 / 免疫シナプス / raft / アゴニストペプチド / ヒストンアセチル化 / Th1・Th2 / rap-1 / インテグリン / SPA-1 / Gab-2 / CTLA-4 / Rap-1 / Spa-1 / Th1 / Th2 / カルシニューリン / シグナルクロストーク / 部分アゴニスト

Research Abstract

本研究においては、T細胞が抗原を認識して活性化され、機能分化や機能発現する際の、抗原ペプチド、抑制性アダプター分子、G蛋白、MAPKやCN系、などによるTCRシグナル伝達の制御を解析し、Th1/Th2分化やアナジー誘導などのT細胞機能調節系を解明した。
1)部分アゴニストペプチドによるT細胞応答は、ZAP70/LAT/SLP76などを活性化せずに、PKCm/Ras/B-raf/Erkの活性化に依存した増殖を誘導することが判明した。CLIPペプチドを置換するライブラリーを用いたT細胞エピトープの同定法を開発し、I型糖尿病関連自己抗原を同定した。
2)ラフト局在分子CbpがCbp-EBP50-ERM-Actinの会合によって細胞骨格系にアンカーし、ラフトの細胞膜状の可動性を調節するシステムが存在し、免疫シナプス形成をも制御していることを見いだした。Gab2がTCRシグナル系ではフィードバック抑制を担うアダプター分子であり、その抑制機構を明らかにした。細胞表面抑制分子CTLA-4がT細胞分化選択でも抑制機能を有することを示した。
3)G蛋白Rap1がインテグリンの主要な活性化因子であることを見つけ、そのGAPであるSPA-1はRap1の制御を通して、免疫シナプス形成・活性化様式の決定に関わることを明らかにした。SPA-1欠損マウスを作製し、免疫応答不全とともに慢性骨髄性白血病を発症することから、SPA-1が癌抑制遺伝子であることを示した。
4)Th2への分化に必要なTCRシグナル伝達系を解析し、Lck,Ras/Erk,Calcineurinの強い活性化がTh2分化に必要なことを見いだした。実際にマウス喘息モデルにおいて、Th2のRas/MAPKによる亢進と、SHP-1による抑制を証明した。また、Th2分化におけるGATA3によるクロマチンリモデリングの分子機構を解明し、遺伝子発現制御に関与するポリコーム遺伝子がTh2分化に必須であることを明らかにした。

Research Products

• [Publications] Itoh, K.: "Negative regulation of immune synapse formation by anchoring lipid raft to cytoskeleton through Cbp-EBP50-ERM assembly"J. Immunol.. 168・2. 541-544 (2002)
• [Publications] Azeredo da Silveira, S.: "Complement activation selectively potentiates the pathogenicity of the IgG2b and IgG3 isotypes of a high affinity anti-erythrocyte autoantibody"J. Exp. Med.. 195・6. 665-672 (2002)
• [Publications] Ioan-Facsinay, A.: "FcγRI (CD64) contributes substantially to severity of arthritis, hypersensitivity responses and protection from bacterial infection"Immunity. 16・3. 391-402 (2002)
• [Publications] Yokosuka, T.: "Predominant role of T cell receptor α chain in forming preimmune TCR repertoire revealed by clonal TCR reconstitution system"J. Exp. Med.. 195・8. 991-1001 (2002)
• [Publications] Murata, Y.: "The conversion of redox status of peritoneal macrophages during pathological progression of spontaneous inflammatory bowl disease in Janus family tyrosine kinase 3-/-and IL-2Rγ-/-mice"Int. Immunol.. 14・6. 627-636 (2002)
• [Publications] Redegeld, F.A.: "Immunoglobulin free light chains elicit immediate hypersensitivity-like responses"Nat. Med.. 8・7. 694-701 (2002)
• [Publications] Snell, D.C.: "Differential effects of reduced glycoprotein VI levels on activation of murine platelets by glycoprotein VI ligands"Biochem. J.. 368・Pt 1. 293-300 (2002)
• [Publications] Tarzi, R.M.: "Nephrotoxic nephritis is mediated by Fcγ receptors on circulating leukocytes and not intrinsic renal cells"Kidney Int.. 62・6. 2087-2096 (2002)
• [Publications] Matsumoto, K.: "Fc receptor-independent development of autoimmune glomerulonephritis in lupus-prone MRL lpr mice"Arthritis. Rheum.. 48・2. 486-494 (2003)
• [Publications] Yamasaki, S.: "Gads/Grb2-mediated association with LAT is critical for the inhibitory function of Gab2 in T cells"Moll. Cell. Biol. (in press).
• [Publications] Arase, N.: "IgE-mediated activation of NK cells through FcγRIII"J. Immunol. (in press).
• [Publications] Saito, T.: "Negative feedback of T cell activation through inhibitory adaptors and costimulatory receptors"Immunol. Rev.. (in press).
• [Publications] Nakayama, T.: "The generation of mature, single-positive thymocytes in vivo is dysregulated by CD69 blockade or overexpression"J. Immunol.. 168. 87-94 (2002)
• [Publications] Suto, A.: "CD4^+ CD25^+ T-cell development is regulated by at least 2 distinct mechanisms"Blood. 99. 555-560 (2002)
• [Publications] Gonzalez-Aseguinolaza, G.: "Natural killer T cell ligand α-galactosylceramide enhances protective immunity induced by malaria vaccines"J. Exp. Med.. 195. 617-624 (2002)
• [Publications] Shibata, Y.: "Ras activation in T cells determines the development of antigen-induced airway hyperresponsiveness and eosinophilic inflammation"J. Immunol.. 196. 2134-2140 (2002)
• [Publications] Yamashita, M.: "Identification of a conserved GATA3 response element upstream proximal of the IL-13 gene locus"J. Biol. Chem.. 277. 42399-42408 (2002)
• [Publications] Kamata, T.: "src homology 2 domain-containing tyrosine phosphatase SHP-1 controls the development of allergic airway inflammation"J. Clin. Inv.. 111. 109-119 (2003)
• [Publications] Iwai, Y.: "Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade"Proc Natl Acad Sci U S A. 99・19. 12293-12297 (2002)
• [Publications] Ishida, M.: "Differential expression of PD-L1 and PD-L2, ligands for an inhibitory receptor PD-1, in the cells of lymphohematopoietic tissues"Immunol Lett.. 84・1. 57-62 (2002)
• [Publications] Katagiri, K.: "Rap1 functions as a key regulator of T-cell and antigen-presenting cell interactions and modulates T-cell responses"Mol Cell Biol. 22・4. 1001-1015 (2002)
• [Publications] Yamanaka, K.: "Identification of the ubiquitin-protein ligase that recognizes oxidized IRP2"Nat Cell Biol.. (in press).
• [Publications] Su, L.: "AF-6 controls integrin-mediated cell adhesion by regulating Rap1 activation through the specific recruitment of Rap1GTP and SPA-1"J Biol Chem.. (in press).
• [Publications] Monji, M.: "Head and neck cancer antigens recognized by the humoral immune system"Biochem. Biophys. Res. Comm. 294. 734-741 (2002)
• [Publications] Nakatsura, T.: "Cellular and humoral immune responses to a human pancreatic cancer antigen, clp, originally defined by the SEREX method"Eur. J. Immunol.. 32. 826-836 (2002)
• [Publications] Uemura, Y.: "Systematic analysis of the combinatorial nature of epitopes recognized by TCR leads to identification of mimicry epitopes for GAD65 specific TCRs"J. Immunol.. 170. 947-960 (2003)
• [Publications] Uemura, Y: "Specificity, degeneracy, and molecular mimicry in antigen recognition by HLA-class II restricted T cell receptors ; Implications for clinical medicine"Modern Rheumatology. (in press).
• [Publications] Soejima, H: "The preference to a Th1-type response in patients with coronary spastic angina"Circulation. (in press).
• [Publications] Senju, S.: "Generation and genetic modification of dendritic cells derived from mouse embryonic stem cells"Blood. (in press).
• [Publications] Irie, A.: "Unique T cell proliferation dependent on PKCμ activation with impaired Zap-70 phosphorylation in recognition of over-expressed HLA-DR/partially agonistic peptide"Eur. J. Immunol.. (in press).
• [Publications] Arase, H.: "Negative regulation of expression and function of Fcγ RIII by CD3ζ in murine NK cells"J. Immunol.. 166・1. 21-25 (2001)
• [Publications] Tomita, K.: "Cytokine-indepeudent Jak3 activation upon TCR stimulation through direct association of Jak3 and the TCR complex"J. Biol. Chem.. 276・27. 25378-25385 (2001)
• [Publications] Arase, H.: "The mouse NK cell-associated antigen recognized by DX5 mAb is CD49β(α_2 integrin, VLA-2)"J. Immunol.. 167・3. 1141-1144 (2001)
• [Publications] Takeuchi, A.: "E2A and HEB activate the pre-TCRα promoter during immature T cell development"J. Immunol.. 167・4. 2157-2163 (2001)
• [Publications] Yamasaki, S.: "Docking protein Gab2 is phosphorylated by ZAP-70 and negatively regulates T cell recepor signaling by recruitment of inhibitory molecules"J. Biol. Chem.. 276・48. 45175-45183 (2001)
• [Publications] Itoh, K.: "Negative regulation of immune synapse formation by anchoring lipid raft to cytoskeleton through Cbp-EBP50-ERM assembly"J. Immunol.. 168・2. 541-544 (2002)
• [Publications] Azeredo da Silveira, S.: "Complement activation selectively potentiates the pathogenicity of the IgG2b and IgG3 isotypes of a high-affinity anti-erythrocyte autoantibody"J. Exp. Med.. (In press).
• [Publications] Ioan-Facsinay, A.: "FcγRI(CD64) contributes substantially to severity of arthritis, hypersensitivity responses and protection from bacterial infection"Immunity. (In press).
• [Publications] Yokosuka, T.: "Predominant role of TCRα chain in forming pre-immune TCR repertoire revealed by clonal TCR reconstitution system"J. Exp. Med.. (In press).
• [Publications] Kimura, M.: "Regulation of Th2 cell differentiation by mel-18, a mammalian polycomb group gene"Immunity. 15. 275-287 (2001)
• [Publications] Sharif, S.: "Activation of natural killer T cells by α-galactosylceramide treatment prevents the onset and recurrence of autoimmune Type 1 diabetes"Nature Medicine. 7. 1057-1062 (2001)
• [Publications] Nakayama, T.: "The generation of mature, single-positive thymocytes in vitro is dysregulated by CD69 blockade or overexpression"J. Immunol.. 168. 87-94 (2002)
• [Publications] Suga, K.: "CD98 induces LFA-1-mediated cell adhesion in lymphoid cells via activation of Rap-1"FEBS Lett.. 489. 249-253 (2001)
• [Publications] Nishimura, H.: "Autoimmune dilated cardiomyopathy in PD-1 receptor deficient mice"Science. 291. 319-322 (2001)
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• [Publications] Okuda, H.: "The Von Hippel-Lindau (VHL) tumor suppressor protein mediates ubiquitination of activated atypical protein kinase C"J. Biol Chem.. 276. 43611-44361 (2001)
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• [Publications] Nishimura, Y: "Molecular and cellular analyses of HLA class II-associated susceptibility to autoimmune diseases in the Japanese population"Modern Rheumatol.. 11. 103-112 (2001)
• [Publications] Inoue, R.: "Identification of β lactoglobulin-derived peptides and class II HLA molecules recognized by T Cells from patients with milk allergy"Clin. Exp. Immunol.. 31. 1126-1134 (2001)
• [Publications] Minohara, M.: "Differences between T cell reactivities to major myelin protein-derived peptides in opticospinal and conventional forms of multiple sclerosis and healthy controls"Tissue Antigens. 57. 447-456 (2001)
• [Publications] Yamada, K.: "Identification of a novel autoantigen UACA in patients with panuveitis"Biochem. Biophys. Res. Comm.. 280. 1169-1176 (2001)
• [Publications] Maeda, A.: "Identification of human antitumor cytotoxic T lymphocytes epitopes of recoverin, cancer-associated retinopathy antigen, to achieve a clinical better prognosis in a paraneoplastic syndrome"Eur. J. Immunol.. 31. 563-572 (2001)
• [Publications] Nakatsura, T.: "Gene cloning of immunogenic antigens over-expressed in pancreatic cancer"Biochem. Biophys. Res. Comm.. 281. 936-944 (2001)
• [Publications] Fujii, S.: "Establishment of an expression cloning system for CD4+ T cell epitopes"Biochem. Biophys. Res. Comm.. 284. 1140-1147 (2001)
• [Publications] Masuda, M.: "Identification and immunocytochemical analysis of DCNP1, a dendritic cell-associated nuclear protein"Biochem. Biophys. Res. Comm.. (in press).
• [Publications] Nakatsura, T.: "Cellular and humoral immune responses to a human pancreatic cancer antigen, CLP, originally defined by the SEREX method"Eur. J. Immunol.. (in press).
• [Publications] Suzuki,K.: "Janus kinase 3 (Jak 3) is essential for common cytokine receptor γ chain(γc)-dependen signaling : comparative analysis of γc, Jak 3, and γc and Jak 3 double deficient mice."Int.Immunol.. 12. 123-132 (2000)
• [Publications] Fossati-Jimack,L.: "Markedly different pathogenicity of four immunoguloblin Gisotype-switch variants o an antierythrocyte autoantibody is based on their capacity to interact in vivo with low-affinity Fcγ receptor III."J.Exp.Med.. 191. 1293-1302 (2000)
• [Publications] Hamano,Y.: "Immune complex and Fc receptor-mediated augmentation of antigen presentation for in vivo helper T cell responses."J.Immunol.. 164. 6113-6119 (2000)
• [Publications] Nakatsu,F.: "A di-leucine signal in the ubiquitin moiety : possible involvement in ubiquitination-mediated endocytosis."J.Biol.Chem.. 275. 26213-26219 (2000)
• [Publications] Tsujino,S.: "Differential requirement of the cytoplasmic subregions of γc chain in T cell development and function."Proc.Natl.Acad.Sci.USA. 97. 10514-10519 (2000)
• [Publications] Suzuki,K.: "Role of common cytokine receptor γ chain (γc)-and Jak3-dependent signaling in the proliferation and survival of murine mast cells."Blood. 96. 2172-2180 (2000)
• [Publications] Iida,T.: "Regulation of cell surface expression of CTLA-4 by secretion of CTLA-4-containing lysosmes upon activation of CD4^+ T cells."J.Immunol.. 165. 5062-5068 (2000)
• [Publications] Okazaki,Y.: "Cell surface expression of calnexin, a molecular chaperone in the endoplasmic reticulum."J.Biol.Chem.. 275. 35751-35758 (2000)
• [Publications] Watanabe,N.: "The quantity of TCR signaling determines positive selection and commitment of cells."J.Immunol.. 165. 6252-6261 (2000)
• [Publications] Arase,H.: "Negative regulation of expression and function of Fcγ RIII by CD3ζ in murine NK cells."J.Immunol.. 166. 21-25 (2001)
• [Publications] Okada,T.: "BCAP : The tyrosine kinase substrate that connects B cell receptor to phosphoinositide 3-kinase activation."Immunity. 13. 817-827 (2000)
• [Publications] Ishiai,M.: "Involvement of LAT, Gads, and Grb2 in compartmentation of SLP-76 to the plasma membrane."J.Exp.Med.. 192. 847-856 (2000)
• [Publications] Hashimoto,A.: "Essential role of phospholopase C-γ 2 in B cell development and function."J.Immunol.. 165. 1738-1742 (2000)
• [Publications] Yasuda,T.: "Cb1 suppresses B cell receptor-mediated phospholipase C (PLC)-γ 2 activation by regulating B cell linker protein-PLC-γ 2 binding."J.Exp.Med.. 191. 641-650 (2000)
• [Publications] Kurosaki,T.: "BLNK : Connecting Syk and Btk to calcium signals."Immunity. 12. 1-5 (2000)
• [Publications] Kurosaki,T.: "Functional dissection of BCR signaling pathways"Curr.Opin.Immunol.. 12. 276-281 (2000)
• [Publications] Liou,J.: "HPK1 is activated by lymphocyte antigen receptors and negatively regulates AP-1."Immunity. 12. 399-408 (2000)
• [Publications] Wong,J.: "Functional complementation of BLNK by SLP-76 and LAT linker proteins."J.Biol.Chem.. 275. 33116-33122 (2000)
• [Publications] Pogue,S.L.: "B cell antigen receptor-induced activation of Akt promotes B cell survival and is dependent on Syk kinase."J.Immunol.. 165. 1300-1306 (2000)
• [Publications] Grishin,A.: "Involvement of Shc and Cbl-PI 3-kinase in Lyn-dependent proliferative signalling pathways for G-CSF."Oncogene. 19. 97-105 (2000)
• [Publications] Burrows,P.D.: "Activation of self-reactive B cells and autoimmune diseases."Reviews in Immunogenetics. 2. 38-51 (2000)
• [Publications] Yayoshi-Yamamoto,S.: "FRL, a novel member of Formin-related proteins, that binds to Rac and regulates cell motility and survival of macrophages."Mol.Cet.Biol.. 20. 6872-6881 (2000)
• [Publications] Ochi,H.: "Negative regulation of BCR-mediated signaling in B-1 cells through CD5 and Ly49 coreceptors via Lyn kinase activity."Int.Immunol.. 12. 1417-1423 (2000)
• [Publications] Kondo,S.: "Suppression of insulitis and diabetes in B cell-deficient mice treated with streptozocin : b cells are essential for the T cell receptor clonotype spreading of islet-infiltrating T cells."Int.Immunol.. 12. 1075-1083 (2000)
• [Publications] Kobayashi,T.: "Abnormal functional and morphological regulation of the gastric mucosa in histamine H2 receptor-dificient mice."J.Clin.Invest.. 105. 1741-1749 (2000)
• [Publications] Marguet,D.: "Enhanced insulin secretion and improved glucose tolerance in mice lacking CD26."Proc.Natl.Acad.Sci.USA. 97. 6874-6879 (2000)
• [Publications] Sonoda,K.: "The clinical significance of tumor-associated antigen RCAS1 expression in the normal, hyperplastic, and malignant uterine endometrium"Gynecol.Oncol.. 79. 424-429 (2000)
• [Publications] Iwasaki,T.: "The expression of human tumor-associated antigen RCAS1 and the prognostic significance in lung cancer."Int.J.Cancer. 89. 488-493 (2000)
• [Publications] Nakahara,H.: "Vascular permeability in allergic conjunctivitis in mice lacking histamine H1 receptors."Eur.J.Pharmacol.. 409. 313-317 (2000)
• [Publications] Izumizaki,M.: "Lack of temperature-induced polypnea in histamine H1 receptor-deficient mice."Neurosci.Lett. 284. 139-142 (2000)
• [Publications] Mobarakeh,J.I.: "Role of histamine H1 receptor in pain perception : a study of the receptor gene knockout mice."Eur.J.Pharmacol.. 391. 81-89 (2000)
• [Publications] Ohshima,K.: "Expression of RCAS1 and FasL in human trophoblasts and uterine glands during pregnancy ; the possible role in immune privilege."Clin.Exp.Immunol.. (In press).
• [Publications] Masaki,T.: "Targeted disruption of histamine H1 receptor attenuated regulatory effect of leptin on feeding behavior, adiposity and UCP family expression in mice."Diabetes. (In press).
• [Publications] Masaki,T.: "Central infusion of hitamine reduced fat accumulation and increased UCP family expression in leptin resistant obese mice."Diabetes. (In press).