| Project/Area Number |
09671086
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
内分泌・代謝学
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| Research Institution | Okinaka memorial institute for medical research |
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Principal Investigator |
NAKANISHI Koji Okinaka memorial institute for medical research, staff, 研究員 (80211423)
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| Project Period (FY) |
1997 – 1999
|
| Project Status |
Completed (Fiscal Year 1999)
|
| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | insulin-dependent diabetes mellitus / MHC molecule / antigenic peptide |
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| Research Abstract |
Susceptibility to insulin-dependent diabetes mellitus (IDDM) is conferred by MHC. MHC molecule presents antigenic peptide to T cells. The motif of this antigenic peptide is influenced by polymorphism of MHC molecules. The aim of this study is to identify islet-derived antigenic peptide presented by IDDM-susceptible MHC molecules.
From peripheral blood of an IDDM patient, B cell line was generated and pulsed with debris of fatal islet cell line. HLA class II molecules was obtained from this B cell line by affinity chromatography using anti-HLA DR and anti-HLA DQ monoclonal antibody. Furthermore, antigenic peptide was eluted from MHC groove using trifluoroacetic acid and subjected to HPLC.
Comparison of HPLC patterns of antigenic peptides with or without pulse of debris of fatal islet cell line identified 3 peaks specific for pulse with fatal islet. Sequencing of these peptides identified one peptide consisted of 14 amino acids. Homology search of this peptide also identified native protein.
cDNA of this protein was obtained by RT-PCR from fatal islet cell line. This cDNA was cloned into PCR 2.1 vector and expression vector pTrcHisA. Protein expression is now trying in E. coli system.
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