1994 Fiscal Year Annual Research Report

Tリンパ球特異的チロシンキナーゼItkの細胞増殖分化における役割

Research Project

Project/Area Number	06281202
Research Institution	Tohoku University
Principal Investigator	中村正孝東北大学, 医学部, 助教授 (30180392)
Co-Investigator(Kenkyū-buntansha)	大保和之東北大学, 医学部, 助手 (70250751) 竹下敏一東北大学, 医学部, 助教授 (60212023)
Keywords	T細胞 / チロシンリン酸化 / Itk / γ鎖 / IL‐2
Research Abstract	ItkはT細胞に特異的に発現しているチロシンキナーゼで、その機能、特にT細胞の増殖分化での役割の解明が待たれている。Itkの機能を解明するために、まず、Itkに対する抗体を樹立した。抗原として、大腸菌で産生したGSTとヒトItkN末端の融合蛋白を用い、ヒトItkに対する単クロン抗体(TUK35)を作成した。本抗体はItkを免疫沈降することができるばかりでなく、ウエスタンブロットでもItkを検出できる。どのような刺激によりItkが活性化されるか調べるために、T細胞を種々の刺激で活性した後、Itkのチロシンリン酸を検討した。抗TCR/CD3あるいはCD28抗体で刺激すると、刺激後早期にItkのチロシンリン酸化が認められた。さらに、抗体による免疫沈降物のなかにFynが含まれており、ItkとFynが細胞内で会合していることが示唆された。転写促進因子として知られるHTLV‐1Taxが、Itkの発現を転写レベルで促進することを証明した。機能的なIL‐2受容体は少なくともα鎖、β鎖、γ鎖の3種類のサブユニットより成る。IL‐2によるT細胞増殖に関与するチロシンキナーゼを同定するため、IL‐2受容体に含まれるチロシンキナーゼを検討した。抗β鎖抗体による免疫沈降物中にJAK1チロシンキナーゼ分子が含まれ、抗γ鎖抗体の免疫沈降物にJAK3が含まれていることを見い出した。これらの検出はIL‐2に依存しない。また、IL‐2刺激により、JAK1、JAK3ともチロシンリン酸化され、IL‐2刺激によりこれらのチロシンキナーゼが活性化されるものと考えられる。さらに、β鎖、γ鎖のサブユニットの変異体を用いた実験から、JAK1との会合、活性化にはβ鎖のセリン領域が、JAK3との会合、活性化にはγ鎖のSH2様領域が必要であることも明らかにした。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] Kondo,M.: "Functional participation of the IL‐2 receptor γ chain in IL‐7 receptor complexes." Science. 263. 1453-1454 (1994)
[Publications] Tanaka,N.: "Physical association of JAK1 and JAK2 tyrosine kinases with the interleukin 2 receptor β and γ chains." Proc.Natl.Acad.Sci.USA. 91. 7271-7275 (1994)
[Publications] Asao,H.: "Interleukin 2‐induced activation of JAK3:possible involvement in signal transduction for c‐myc enduction and cell proliferation." FEBS Letters. 351. 201-206 (1994)
[Publications] Fujii,M.: "Serum response factor has functional roles both in indirect binding to the CArG box and in the transcriptional activation function of human T‐cell leukemia virus type I Tax." J.Virol.68. 7275-7283 (1994)
[Publications] Uchijima,M.: "Tax protein of HTLV‐1 and‐2 induce expression of the gene encoding erythroid‐potentiating activity(tissue inhibitor of metalloproteinases‐1:TIMP‐1)." J.Biol.Chem.269. 14946-14950 (1994)
[Publications] Tsuchiya,H.: "Two distinct regions from a functional activation domain of the HTLV‐1 trans‐activator Tax1." Oncogene. 9. 337-340 (1994)
[Publications] Fujita,K.: "Enhancement of T cell receptor Signaling of Tumor‐infiltrating Lymphocytes by Retrovirally Mediated fyn Gene Transduction." Jpn.J.Cancer.Res.85. 1073-1079 (1994)
[Publications] Akagi,T.: "Increased protein tyrosine‐phosphorylation in primary T‐cells transduced wity Taxl of Human T‐cell leukemia virus type I." FEBS Letters.358. 34-38 (1995)
[Publications] Hirai,H.: "Tax protein of human T‐cell leulemia virus type I binds to the ankyrin motifs inhibitory fant kB and induces nuclear translocation of transcription factor NF‐kB proteins for transcriptional activation." Proc.Natl.Acad.Sci.USA.91. 3584-3588 (1994)
[Publications] Fujimoto,T.: "Novel isoforms of human cyclic AMP‐responsive element modulator(hCREM)mRNA." J.Biochem.115. 298-303 (1994)
[Publications] Yamada,T.: "The HTLV‐1 Tax protein induces apoptosis which is blocked by the Bcl‐2 protein." J.Virol.68. 3374-3379 (1994)
[Publications] Inoue,H.: "Suppression of the transformed phenotype in hybrids of human T‐cell leukemia virus type I Tax‐transformed rat fibroblasts and normal human fibroblasts." Exp.Cell Res.215. 68-74 (1994)
[Publications] Nishimura,M.: "Demonstration of human T lymphotropic virus type 1 (HTLV‐1)‐specific T cell responses from seronegative and polymerase chain reaction‐negative persons exposed to HTLV‐1." J.Infect Dis.170. 334-338 (1994)
[Publications] Uchiyama,T.: "Pathophysiology of adult T cell leukemia cells‐cell growth characteristics‐‐ In"Adult T cell leukemia"(ed.by K.Takatsuki)" Oxford University Press,Oxford. 181-203 (1994)
[Publications] Takaori‐K.: "Detection of homing,proliferation.and infiltration sites of adult Tcellleukemiacells in severe combined immunodeficiency mice using radiometric technques." Jpn.J.Cancer Res. (in press). (1995)
[Publications] Tateno,Y.: "Genetic and functional variabilities in the human immunodeficiency virus type 1 isolated from the same blood samples with and without CD8+cells." Immunology & Infectious Disease. 4. 41-46 (1994)
[Publications] Kitamura,M.: "Expression of an endogenous retrovial gene product in human placenta." Int.J.Cancer. 58. 836-840 (1994)
[Publications] Seto,K.: "A rat model of HTLV‐J infection.Development of chronic progressive myeloneuropathy in seropositive WKAH rats and related apoptosis." Acta Neuropahol. (in press).
[Publications] Sakai,R.: "A novel signaling molecule,p130,forms stable complexes in vivo with v‐Crk and v‐Src in a tyrosine phosphorylation‐dependent manner." EMBOJ.13. 3748-3756 (1994)
[Publications] Okumura,N.: "Vasoactive intestinal peptide induces differentiation and MAP kinase activation in PC12h cells" J.Biochem.115. 304-308 (1994)
[Publications] Sato,S.: "IL‐5 receptor‐mediated tyrosine phosphorylation of SH2/SH3‐containing proteins and activation of Bruton's tyrosine and Janus 2 kinases." J.Exp.Med.180. 2101-2111 (1994)
[Publications] Tomida,S.: "Intercellular adhesion molecule‐1 and leukocyte function‐associated antigen‐1 are involved in protection mediated by CD3+TCRab‐T cells at the early stage after infection with Listeria monocytogenes in rats." Int.Immunol.6. 955-961 (1994)
[Publications] Aoki,Y.: "Clonal expansion but lack of subsequent clonal deletion of bacterial superantigen‐reactive T cells in murine retroviral infection." J.Immunol.153. 3611-3621 (1994)

1994 Fiscal Year Annual Research Report

Tリンパ球特異的チロシンキナーゼItkの細胞増殖分化における役割

Principal Investigator

中村 正孝 東北大学, 医学部, 助教授 (30180392)

Research Products

[Publications] Kondo,M.: "Functional participation of the IL‐2 receptor γ chain in IL‐7 receptor complexes." Science. 263. 1453-1454 (1994)

[Publications] Tanaka,N.: "Physical association of JAK1 and JAK2 tyrosine kinases with the interleukin 2 receptor β and γ chains." Proc.Natl.Acad.Sci.USA. 91. 7271-7275 (1994)

[Publications] Asao,H.: "Interleukin 2‐induced activation of JAK3:possible involvement in signal transduction for c‐myc enduction and cell proliferation." FEBS Letters. 351. 201-206 (1994)

[Publications] Fujii,M.: "Serum response factor has functional roles both in indirect binding to the CArG box and in the transcriptional activation function of human T‐cell leukemia virus type I Tax." J.Virol.68. 7275-7283 (1994)

[Publications] Uchijima,M.: "Tax protein of HTLV‐1 and‐2 induce expression of the gene encoding erythroid‐potentiating activity(tissue inhibitor of metalloproteinases‐1:TIMP‐1)." J.Biol.Chem.269. 14946-14950 (1994)

[Publications] Tsuchiya,H.: "Two distinct regions from a functional activation domain of the HTLV‐1 trans‐activator Tax1." Oncogene. 9. 337-340 (1994)

[Publications] Fujita,K.: "Enhancement of T cell receptor Signaling of Tumor‐infiltrating Lymphocytes by Retrovirally Mediated fyn Gene Transduction." Jpn.J.Cancer.Res.85. 1073-1079 (1994)

[Publications] Akagi,T.: "Increased protein tyrosine‐phosphorylation in primary T‐cells transduced wity Taxl of Human T‐cell leukemia virus type I." FEBS Letters.358. 34-38 (1995)

[Publications] Hirai,H.: "Tax protein of human T‐cell leulemia virus type I binds to the ankyrin motifs inhibitory fant kB and induces nuclear translocation of transcription factor NF‐kB proteins for transcriptional activation." Proc.Natl.Acad.Sci.USA.91. 3584-3588 (1994)

[Publications] Fujimoto,T.: "Novel isoforms of human cyclic AMP‐responsive element modulator(hCREM)mRNA." J.Biochem.115. 298-303 (1994)

[Publications] Yamada,T.: "The HTLV‐1 Tax protein induces apoptosis which is blocked by the Bcl‐2 protein." J.Virol.68. 3374-3379 (1994)

[Publications] Inoue,H.: "Suppression of the transformed phenotype in hybrids of human T‐cell leukemia virus type I Tax‐transformed rat fibroblasts and normal human fibroblasts." Exp.Cell Res.215. 68-74 (1994)

[Publications] Nishimura,M.: "Demonstration of human T lymphotropic virus type 1 (HTLV‐1)‐specific T cell responses from seronegative and polymerase chain reaction‐negative persons exposed to HTLV‐1." J.Infect Dis.170. 334-338 (1994)

[Publications] Uchiyama,T.: "Pathophysiology of adult T cell leukemia cells‐cell growth characteristics‐‐ In"Adult T cell leukemia"(ed.by K.Takatsuki)" Oxford University Press,Oxford. 181-203 (1994)

[Publications] Takaori‐K.: "Detection of homing,proliferation.and infiltration sites of adult Tcellleukemiacells in severe combined immunodeficiency mice using radiometric technques." Jpn.J.Cancer Res. (in press). (1995)

[Publications] Tateno,Y.: "Genetic and functional variabilities in the human immunodeficiency virus type 1 isolated from the same blood samples with and without CD8+cells." Immunology & Infectious Disease. 4. 41-46 (1994)

[Publications] Kitamura,M.: "Expression of an endogenous retrovial gene product in human placenta." Int.J.Cancer. 58. 836-840 (1994)

[Publications] Seto,K.: "A rat model of HTLV‐J infection.Development of chronic progressive myeloneuropathy in seropositive WKAH rats and related apoptosis." Acta Neuropahol. (in press).

[Publications] Sakai,R.: "A novel signaling molecule,p130,forms stable complexes in vivo with v‐Crk and v‐Src in a tyrosine phosphorylation‐dependent manner." EMBOJ.13. 3748-3756 (1994)

[Publications] Okumura,N.: "Vasoactive intestinal peptide induces differentiation and MAP kinase activation in PC12h cells" J.Biochem.115. 304-308 (1994)

[Publications] Sato,S.: "IL‐5 receptor‐mediated tyrosine phosphorylation of SH2/SH3‐containing proteins and activation of Bruton's tyrosine and Janus 2 kinases." J.Exp.Med.180. 2101-2111 (1994)

[Publications] Tomida,S.: "Intercellular adhesion molecule‐1 and leukocyte function‐associated antigen‐1 are involved in protection mediated by CD3+TCRab‐T cells at the early stage after infection with Listeria monocytogenes in rats." Int.Immunol.6. 955-961 (1994)

[Publications] Aoki,Y.: "Clonal expansion but lack of subsequent clonal deletion of bacterial superantigen‐reactive T cells in murine retroviral infection." J.Immunol.153. 3611-3621 (1994)

中村正孝東北大学, 医学部, 助教授 (30180392)