| Project/Area Number |
06454261
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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 | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Shinshu University |
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Principal Investigator |
KIYOSAWA Kendo Shinshu University School of Medicine, Professor, 医学部, 教授 (30020829)
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| Co-Investigator(Kenkyū-buntansha) |
IMAI Haruhiko Shinshu University School of Medicine, Instructor, 医学部, 助手
HASEGAWA Akira Research and Development Laboratories, TONEN Corporation, Head, 総合研究所, 課長
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1996: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1994: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | autoimmune hepatitis / asialoglycoprotein receptor / autoantibody / molecular biology |
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| Research Abstract |
Reverse transcription and polymerase chain reaction was applied to isolate cDNAs encoding asialoglycoprotein receptor (ASGPR) H1 and L-H2 from human liver mRNAs. Using these cDNAs, recombinant ASGPR protein was expressed in E.coli. Rabbit antisera specific to human ASGPR H1 or L-H2 was obtained by immunizing these recombinant proteins. Enzyme-linked immunosorbent assay (ELISA) using bacterial recombinant antigen was developed to measure serum antibodies to ASGPR.Sera from autoimmune hepatitis, primary biliary cirrhosis, alcoholic liver disease, chronic hepatitis C,chronic hepatitis B,systemic lupus erythematosus, and healthy subjects were tested. High OD value was obtained almost exclusively in autoimmune hepatitis and chronic hepatitis C.The OD value of anti-ASGPR antibodies detected by ELISA using recombinant antigen was correlated with anti-ASGPR antibody titers that were determined by ELISA using human liver antigen. By using extracellular domain of ASGPR protein in ELISA,similar sensitivity to pick-up anti-ASGPR antibodies was demonstrated in autoimmune hepatitis and less reactivity was detected in chronic hepatitis C.It was suggested that main epitope which was reactive with antibodies in autoimmune hepatitis could be located on the extracellular domain of the ASGPR protein. Lymphocytes from peripheral blood mononuclear cells in a patient with autoimmune hepatitis showed a stimulation index as 4 in lymphocyte stimulation test with recombinant ASGPR antigen. There could exist a certain class of lymphocytes reactive with human ASGPR in autoimmune hepatitis patients.
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