1999 Fiscal Year Final Research Report Summary
Regulation of T cell activity and detection of TCR-binding antigen by recombinant soluble TCR molecules
| Project/Area Number |
10557037
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Kyushu University |
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Principal Investigator |
WATANABE Takeshi Medical Institute of Bioregulation, Kyushu University, Professor, 生体防御医学研究所, 教授 (40028684)
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| Project Period (FY) |
1998 – 1999
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| Keywords | TCR / T cell hybridoma / soluble single Fv TCR / Idiotype / Peptide antigen / autorective T cell / localization of auto-antigen |
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| Research Abstract |
It would be one of promising strategies for the treatment of autoimmune diseases, allergic diseases, hypersensitivity and graft rejection of we could manipulate antigen-specific T cell response. In order to manipulate T cell responses in vivo, we produced a soluble single Fv fragment (scFv) of antigen-specific TCR. Genes encoding for TCRα and TCRβ from a T cell hybridoma, DO11.10 which carries TCR recognizing OVA peptide in context with H-2d class I MHC. The isolated Vα and Vβ DNA were ligated with immunoglobulin gene promoter and enhancer. The construct was expressed in mouse myeloma cells. Thus, we could produced an enough amounts of scFv which specifically binds to OVA peptide in context with H-2d restriction. Molecular weight of scFv was 25 kDa and the molecules are stable in vitro. The scFv maintains the same idiotype specificity as original TCR of DO11.10. In vitro T cell response to OVA peptide was specifically blocked in the presence of this scFv. The scFv bound to H-2d positive antigen-presenting cells when the correct OVA peptide was supplied. We also isolated Vα and Vβ DNA from TCR of the MBP (myelin basic proteins)-specific T cell, which was established from SJL mouse suffering EAE (experimental allergic encephalitis) . Then, we succeeded to produce scFv from the MBP specific TCR. By utilizing these scFv molecules, we are now trying to detect in vivo antigen peptides bound to MHC and to modify the immune response to the antigens. These trials will provide us a clue to treat various immunological diseases.
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