| Project/Area Number |
08557027
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Immunology
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| Research Institution | Kumamoto University |
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Principal Investigator |
NISHIMURA Yasuharu Kumamoto Univ.Grad.Sch.of Med.Sci., Dept.Neurosci.and Immunol., Div.of Immunogenetics, Professor, 大学院・医学研究科, 教授 (10156119)
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| Co-Investigator(Kenkyū-buntansha) |
TSUNOO Hajime Meiji Institute of Health Science, Director, ヘルスサイエンス研究所, 所長
SENJU Satoru Kumamoto Univ.Grad.Sch.of Med.Sci., Dept.Neurosci.and Immunol., Div.of Immunogen, 大学院・医学研究科, 助手 (50274709)
MATSUSHITA Sho Kumamoto Univ.Grad.Sch.of Med.Sci., Dept.Neurosci.and Immunol., Div.of Immunogen, 大学院・医学研究科, 助教授 (50167649)
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| Project Period (FY) |
1996 – 1998
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| Keywords | HLA-class II / Antigenic peptide / Antigen presentation / Invariant chain / Helper T cell / T cell receptor / Gene transfection / Cell library |
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| Research Abstract |
In order to understand pathogenesis of autoimmune diseases, it is important to identify ligands for T cell receptor (TCR) of autoreactive T cells. We developed a new system for delivery of an antigenic peptide to the MHC class II pathway, using the invariant chain (Ii). We designed CLIP-substituted human p33-form us in which either streptococcal M12p55-68 peptide or human 65 kDa glutamic acid decarboxylase (GAD65) p116-129 peptide was substituted for CLIP (class II associated invariant chain peptide). Similar to the wild-type Ii, the CLIP-substituted Ii was associated intracellularly with DR4 molecules. We examined the peptide presenting function of these constructs. Hamster CHO cell transfectants stably expressing M12/CLIP-substituted Ii gene together with HLA-DR4 gene could process and present M12p55-68 to a M12-peptide specific and DR4-restricted CD4^+ T cell clone to induce production of IFN-gamma. Even in the presence of 100 CHO cell transfectants in a well of 96-wells micriculture plate in which 3*10^4 CHO cells were accomodated, the T cell cloneproduced a significant ammount of IFN-gamma. Furthermore, CHO cells expressing stably with HLA-DR53 gene and transiently with GAD6S/CLIP-substituted Ii gene stimulated production of IFN-gamma in a HLA-DR53-restricted and GAD65 autoreactive T cell clone established from a Japanese patient with insulin-dependent diabetes mellitus. These results indicate that the peptides substituted for CLIP of Ii p323 bound to the groove of DR molecules in the same manner as CLIP and they were preferentially presented to the CD4^+ T cell clone in the absence of HLA-DM molecules. This system may prove useful for immunotherapy with DNA vaccines or for construction of a library of antigen presenting cells expressing diverse HLA class-II plus peptide complexes for identification of TCR-ligands and the latter research project is now on going.
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