| Project/Area Number |
60870017
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Experimental pathology
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| Research Institution | Asahikawa Medical College |
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Principal Investigator |
KATAGIRI Makoto Asahikawa Medical College, Professor, 医学部, 教授 (10041823)
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| Co-Investigator(Kenkyū-buntansha) |
IKEDA Hisami Hokkaido Red Cross Blood Center, Director of Research Division, 研究部, 部長 (90091561)
HIGA Toshio Asahikawa Medical College, Assistant Professor, 医学部, 助手 (10133836)
YAKURA Hidetaka Asahikawa Medical College, Associate Professor, 医学部, 助教授 (60166486)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1986: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1985: ¥5,200,000 (Direct Cost: ¥5,200,000)
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| Keywords | HLA antigen / class <II> antigen / class <II> gene / HLA-D region / disease susceptibility / 酵素免疫測定法 / 酵素抗体法 |
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| Research Abstract |
Gene products of HLA-D region (class <II> antigens) are involved in the development of a certain category of autoimmune diseases as an etiologic factor. DNA analysis suggested the presence of extra class <II> antigens in addition to four types of class <II> antigen system (DR, DRw52.53, DQ and DP). For the detection of these class <II> molecules, radioimmunoassay (RIA) instead of cytotoxic assay has been widely used. Since the use of radioisotopes, however, is rather restricted, we have been trying to establish non-radioisotopic assay systems for class <II> molecules in the hope that they can eventually be used in the field of clinical pathology.
We have developed three kinds of enzyme immunoassay (EIA) using peroxidase-labeled anti-mouse Ig antibody or biotinylated anti-mouse Ig antibody and avidin-peroxidase.
1. Solid phase EIA method in which solubilized class <II> molecules are attached to the well as target antigens: Various detergents and buffer systems for the attachment of target molecules have been tested. Deoxycholic acid as detergent and tris-HCl as buffer have yielded most reproducible results. The plates thus prepared can be used for three months if stored at -40゜C. It has also been confirmed that this method can be applied for immunochemical analysis of class <II> molecules.
2. Cellular EIA method in which cells themselves are used as targets: Two versions of this method have been established using either Millititer SV plates (Millipore Corp.) or Immulon 600 flat-bottomed plates (Greiner). The former is convenient in that cells can be washed by aspiration from the bottom of the well, and the latter is advantageous because the plates can be stored for three months at 4゜C once cells are attached to the well. Furthermore, biotin-avidin system has been shown to be more sensitive than RIA in detecting class <II> antigens. We hope that these methods will be used in various fields of medical science.
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