1997 年度実績報告書

TCRシグナルによるT細胞の分化・選択の制御機構

研究課題

研究課題/領域番号	08457102
研究機関	千葉大学
研究代表者	斎藤隆千葉大学, 医学部, 教授 (50205655)
研究分担者	荒瀬尚千葉大学, 医学部, 助手 (10261900) 大野博司千葉大学, 医学部, 助教授 (50233226)
キーワード	Jak3 / TCR / シグナル伝達 / プレTCR複合体 / カルネキシン / DC3γ / ポジティブ・ネガティブ選択 / T細胞分化
研究概要	昨年度の解析により、Jak3がT細胞増設シグナルにのみではなく、T細胞の分化・機能維持にも重要であることが明らかになった。Jak3がTCRシグナルに関与するかを解析した結果、Jak3がTRC刺激によって直接リン酸化されることが判明した。しかも、DC3ζ鎖とJak3をCo-transfectすることによって、Jac3は直接DC3ζ鎖と会合することが明らかになった。キナーゼドメインを欠損した変異Jac3を導入したT細胞では、抗原刺激に伴うIL-2産生が低下したことから、Jac3がTCRシグナルを担う伝達分子であり、IL-2産生を制御していると考えられる。一方、未熟T細胞に発現するプレTCR複合体を解析することによって、プレTCRをカルネキシンと共に、新たな12kd(pTAC12)と会合していることが明らかにした。大量の細胞よりこの蛋白質を精製し、部分アミノ酸配列を決定したところ、CD3γ鎖と同一であることが判明した。そこで5^,RACE法によって、CD3γと配列を共有するcDNAをクローニングしたところ、CD3γの膜貫通部を含むエキソンを欠損するアルタナティスプライシングの産物であることが明らかになった。抗CD3γ鎖抗体を用いることにより、確かに未熟T細胞に検出されるpTAC12が反応することが判明した。また、TCRシグナルによるT細胞分化を解析するため、昨年度はHY特異的TCR-TgとDC3ζ-欠損マウスを用いて解析したが、本年度はペプチド量を変化させることのできなるOVA特異的TCR-TgとDC3ζ欠損マウスを用い、TCRシグナルの強さを低くすることによって、これらのTCRシグナルと選択の量的関係を明らかにすることに成功した。

研究成果
(25件)

すべてその他

すべて文献書誌 (25件)

[文献書誌] Nakano,H.: "Specific interaction beteewn topoisomerase IIβ and CD3ε chain of the Tcell receptor complex." J.Biol.Chem.271. 6483-6489 (1996)
[文献書誌] Takahashi,H.: "Inactivation of Hiv-1 envelope-specific CD8^+ cytotoxicT lymphocytes by free andtiqenic peptide:A self-veto mechanism?" J.Exp.Med.183. 879-889 (1996)
[文献書誌] Arase,H.: "Interferon γ production by natural killer cells and NK1.1^+T cellupon NKR-P1 cross-linking" J.Exp.Med.183. 2391-2396 (1996)
[文献書誌] Sakiyama,H.: "Site-directed mutagenesis of hamster complement Cls:characterization with on active from-speific zntiboody and possible involvemenc of cls in tumorige nicity" Int.J.Cancer. 66. 768-771 (1996)
[文献書誌] van Vugt,M.J: "FcRγ chain is essential for both surface expression and function of human FcγRl(CD64)in vivo" Blood. 87. 3593-3599 (1996)
[文献書誌] Aoe,T.: "Preferntial requierment of CD3ζmediated signals for developmont of immature rather than mature thymocytes" Int.Immunol. 8. 1055-1066 (1996)
[文献書誌] Otsuji,M.: "Oxidative strees by tumor-deried macrophages suppresses the expression of CD3ζ chain of T cellreceptor complex and antigen-specific T cell responses" Prcc.Natl.Acad.Sci.USA. 93. 13319-13124 (1996)
[文献書誌] Saijo,K.: "Crucial role of Jak3 in negative selection of self-reactive T cells" J.Exp.Med. 185. 351-356 (1997)
[文献書誌] Yamazaki,T.: "A shift from negative to positive selection of autoreactive T cells by the reduced level of TCR signal in TVR-transaenic VD3ζ-deficient mice" J.Immunol.158. 1634-1640 (1997)
[文献書誌] Aoe,T.: "Heat shock protein 70 mRNA reflects the severity of ischemia/hypoxia-reperfusion injury in the perfused rat ilver" Crit.Care Med.25. 324-329 (1997)
[文献書誌] Poole,A.: "The Fc receptor γ-chain and the tyrosine kinase Syk are essential for activa-tion of mouse platelets by collagen" EMBO J.16. 2333-2341 (1997)
[文献書誌] Shiratori,T.: "Tyrosine phosphorylation controls internalization of CTLA-4 by regulating its interaction with chlathrin-associated adaptor complex AP-2" Immunity. 6. 583-589 (1997)
[文献書誌] Miyatate,S.: "Induction of interleukin-2 unresponsiveness and down-regulation of the JAK-STAT system upon activation through the T cell receptor" Eur.J.Immunol. 27. 1816-1823 (1997)
[文献書誌] Takase,K.: "A novel 12kd dimer associated with pre-TCR complex and clonotype-independent VD3 complex on immature thymocytes" J.Immunol.159. 741-747 (1997)
[文献書誌] Watanabe,N.: "Th1 and Th2 subsets equally undergo Fas-dependent and independent activa-tion -induced cell daath" Eur.J.Immunol. 27. 1858-1864 (1997)
[文献書誌] Watanabe,H.: "A novel acyl-CoA thioesterase enhances its enzymatic activity by direct binding with HIV nef" Biochem.Biophts.Res.Commun.238. 234-239 (1997)
[文献書誌] Kosugi,A.: "Subunit composition of the pre-T-cell receptor complex analyzed by mono-clonal antibody against the pre-T-cell receptor α chain" Immunology. 91. 618-622 (1997)
[文献書誌] Arase,N.: "Association FcRγ is essential for activation signal through NKR-P1(CD161)NK cells and NK1.1^+T cells" J.Exp.Med. 186. 1957-1963 (1997)
[文献書誌] Wakizaka,K.: "A novel 90kDa tyrosine-phosphorylated protein associated with TCR complex in thymocytes" Eur.J.Immunol.(in press).
[文献書誌] Saito,T.: "Weir´s Handbook of Experimentnal Immunology" Blackwell Science,Inc.Cambridge, (1996)
[文献書誌] Saito,T.: "Weir´s Handbook of Experimentnal Immunology" Blackwell Science,Inc.Cambridge, (1996)Saito,T.:
[文献書誌] "Kinases and phosphatases in lymphocyte and neuronal signaling" Springer-Verlag,Tokyo,Japan, 366 (1997)
[文献書誌] Yamazaki.T.: "Leucocyte typing VI white celldifferentiation antigens" Garland publishing,Inc.,New York and London, 1342 (1997)
[文献書誌] Saijo,K.: "Leucocyte typing VI white celldifferentiation antigens" Garland publishing,Inc.,New York and London, 1342 (1997)
[文献書誌] Saito,T.: "Critical Raviews in Immunology" (in press),

1997 年度 実績報告書

TCRシグナルによるT細胞の分化・選択の制御機構

研究代表者

斎藤 隆 千葉大学, 医学部, 教授 (50205655)

研究成果

[文献書誌] Nakano,H.: "Specific interaction beteewn topoisomerase IIβ and CD3ε chain of the Tcell receptor complex." J.Biol.Chem.271. 6483-6489 (1996)

[文献書誌] Takahashi,H.: "Inactivation of Hiv-1 envelope-specific CD8^+ cytotoxicT lymphocytes by free andtiqenic peptide:A self-veto mechanism?" J.Exp.Med.183. 879-889 (1996)

[文献書誌] Arase,H.: "Interferon γ production by natural killer cells and NK1.1^+T cellupon NKR-P1 cross-linking" J.Exp.Med.183. 2391-2396 (1996)

[文献書誌] Sakiyama,H.: "Site-directed mutagenesis of hamster complement Cls:characterization with on active from-speific zntiboody and possible involvemenc of cls in tumorige nicity" Int.J.Cancer. 66. 768-771 (1996)

[文献書誌] van Vugt,M.J: "FcRγ chain is essential for both surface expression and function of human FcγRl(CD64)in vivo" Blood. 87. 3593-3599 (1996)

[文献書誌] Aoe,T.: "Preferntial requierment of CD3ζmediated signals for developmont of immature rather than mature thymocytes" Int.Immunol. 8. 1055-1066 (1996)

[文献書誌] Otsuji,M.: "Oxidative strees by tumor-deried macrophages suppresses the expression of CD3ζ chain of T cellreceptor complex and antigen-specific T cell responses" Prcc.Natl.Acad.Sci.USA. 93. 13319-13124 (1996)

[文献書誌] Saijo,K.: "Crucial role of Jak3 in negative selection of self-reactive T cells" J.Exp.Med. 185. 351-356 (1997)

[文献書誌] Yamazaki,T.: "A shift from negative to positive selection of autoreactive T cells by the reduced level of TCR signal in TVR-transaenic VD3ζ-deficient mice" J.Immunol.158. 1634-1640 (1997)

[文献書誌] Aoe,T.: "Heat shock protein 70 mRNA reflects the severity of ischemia/hypoxia-reperfusion injury in the perfused rat ilver" Crit.Care Med.25. 324-329 (1997)

[文献書誌] Poole,A.: "The Fc receptor γ-chain and the tyrosine kinase Syk are essential for activa-tion of mouse platelets by collagen" EMBO J.16. 2333-2341 (1997)

[文献書誌] Shiratori,T.: "Tyrosine phosphorylation controls internalization of CTLA-4 by regulating its interaction with chlathrin-associated adaptor complex AP-2" Immunity. 6. 583-589 (1997)

[文献書誌] Miyatate,S.: "Induction of interleukin-2 unresponsiveness and down-regulation of the JAK-STAT system upon activation through the T cell receptor" Eur.J.Immunol. 27. 1816-1823 (1997)

[文献書誌] Takase,K.: "A novel 12kd dimer associated with pre-TCR complex and clonotype-independent VD3 complex on immature thymocytes" J.Immunol.159. 741-747 (1997)

[文献書誌] Watanabe,N.: "Th1 and Th2 subsets equally undergo Fas-dependent and independent activa-tion -induced cell daath" Eur.J.Immunol. 27. 1858-1864 (1997)

[文献書誌] Watanabe,H.: "A novel acyl-CoA thioesterase enhances its enzymatic activity by direct binding with HIV nef" Biochem.Biophts.Res.Commun.238. 234-239 (1997)

[文献書誌] Kosugi,A.: "Subunit composition of the pre-T-cell receptor complex analyzed by mono-clonal antibody against the pre-T-cell receptor α chain" Immunology. 91. 618-622 (1997)

[文献書誌] Arase,N.: "Association FcRγ is essential for activation signal through NKR-P1(CD161)NK cells and NK1.1^+T cells" J.Exp.Med. 186. 1957-1963 (1997)

[文献書誌] Wakizaka,K.: "A novel 90kDa tyrosine-phosphorylated protein associated with TCR complex in thymocytes" Eur.J.Immunol.(in press).

[文献書誌] Saito,T.: "Weir´s Handbook of Experimentnal Immunology" Blackwell Science,Inc.Cambridge, (1996)

[文献書誌] Saito,T.: "Weir´s Handbook of Experimentnal Immunology" Blackwell Science,Inc.Cambridge, (1996)Saito,T.:

[文献書誌] "Kinases and phosphatases in lymphocyte and neuronal signaling" Springer-Verlag,Tokyo,Japan, 366 (1997)

[文献書誌] Yamazaki.T.: "Leucocyte typing VI white celldifferentiation antigens" Garland publishing,Inc.,New York and London, 1342 (1997)

[文献書誌] Saijo,K.: "Leucocyte typing VI white celldifferentiation antigens" Garland publishing,Inc.,New York and London, 1342 (1997)

[文献書誌] Saito,T.: "Critical Raviews in Immunology" (in press),

1997 年度実績報告書

斎藤隆千葉大学, 医学部, 教授 (50205655)