| Project/Area Number |
62480435
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Jichi Medical School |
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Principal Investigator |
KAWAI Tadahi Dept. of Medicine, Jichi Medical School, Professor., 医学部, 教授 (60048957)
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| Co-Investigator(Kenkyū-buntansha) |
ITOH Yoshihisa Dept. of Medicine, Jichi Medical School, Lecturer., 医学部, 講師 (20129026)
KASAHARA Tadashi Dept. of Medicine, Jichi Medical School, Assistant Professor., 医学部, 助教授 (60049096)
向田 直史 自治医科大学, 医学部, 助手 (30182067)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1989: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1988: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥3,900,000 (Direct Cost: ¥3,900,000)
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| Keywords | IgA / Subclass / alpha_1-microglobulin / Multiple myeloma / Enzyme immunoassay / α_1ーマイクログロブリン / 選択的IgA欠損症 / IgA1 / IgA2 / α鎮(α-chain) / IgG / Protein A |
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| Research Abstract |
The present project was initially undertaken to elucidate biological role and clinical significance of IgA subclass in both fundamental and clinical medicine by preparing reliable IgA subclass antigens, their antibodies, and precise assay systems.
We successfully purified IgAl and IgA2 antigen from myeloma sera, prepared mouse monoclonal antibodies, and established an enzyme linked immuno-sorbent assay for both antigens. By making best use of antibodies the studies on alpha-chain disease and alpha_1-microglobulin were facilitated. The summary of the present project was shown below. 1) Purification of IgA1 and IgA2 antigen: Myeloma serum with marked elevation of IgA and depression of other Immunoglobulin (Ig) classes was precipitated with 40-50% ammonium sulfate and passed through anti-alpha-chain antibody affinity chromatography. Purity was confirmed by SDS-PAGE-and IEP.
Identification of its subclass was carried out by monoclonal antibodies by Oxoid and Jackalin. 2) Preparation of specific monoclonal antibodies: A specific high-tetered monoclonal antibody was obtained for IgAl and IgA2 respectively. 3) Analysis of epitope recognition of an anti-IgA1 antibody by IgA1 proteases: After IgAl antigen was digested by proteases (H. Influenza, C. Ramosum), the C-terminal portion of hinge region including Fc portion proved to be recognized by the present antibody. 4) Clinical application: Using an ELISA the serum level of IgA1 and IgA2 was investigated. The average level aged from 10 to 60 was 2.2 mg/ml-in IgA1 and 0.2 mg/ml in IgA2 respectively, therefore IgAl/IgA2 ratio was calculated to 9.4. Of 117 cases with myeloma 104 cases proved to be IgA1 subclass and the rest of 13 was IgA2. Further studies are in progress both fundamentally and clinically which will appear in domestic and international journals.
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