1989 Fiscal Year Final Research Report Summary
Studies on the sub-region of chicken major histocompatibility B locus by using monoclonal antibodies.
Project/Area Number |
63560261
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Research Category |
Grant-in-Aid for General Scientific Research (C)
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Allocation Type | Single-year Grants |
Research Field |
畜産学(含草地学)
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Research Institution | HIROSHIMA UNIVERSITY |
Principal Investigator |
YAMAMOTO Yoshio Fac. Applied Biol. Sci., Assoc. Prof. Hiroshima Univ., 生物生産学部, 助教授 (10032103)
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Co-Investigator(Kenkyū-buntansha) |
MATSUDA Haruo Fac. Applied Biol. Sci., Assoc. Prof. Hiroshima Univ., 生物生産学部, 助教授 (80116863)
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Project Period (FY) |
1988 – 1989
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Keywords | Chicken / MHC / Monoclonal antibody / T lymphocyte / Blood group |
Research Abstract |
The chicken major histocompatibility complex(MHC) was encoded three classes of antigens : B-F(class I), B-L(class II), and a third class, B-G(class IV), which is expressed in erythrocytes and erythroid precusor cells. Chickens of known B genotypes were used as cell donors for the immunization of mice. BALB/c mice were immunized i.p. with 2x10^7 erythrocytes, peripheral lymphocytes, and spleen lymphocytes, respectively, on day 0 and day 14, and received a last i.v. injection of 5x10^6 cells on day 21. Eight fusions were made with spleen cells from mice immunized with B^9B^9 and B^<11>B^<11> erythrocytes(fusions 1 and 2),spleen lymphocytes (fusions 3-6), and peripheral lymphocytes(fusions 7 and 8). Six of 32 hybrydoma were selected as clones which produced a monoclonal antibody. Six Mab were tested their characteristics by flow cytometer(FCM) and serological methods. Four of them were reacted both of erythrocytes arid peripheral lymphocytes. This result suggest that these four Mab has a specificity as B-F sub-region. Two of six Mab were reacted with erythrocytes only. Serological results also suggest that these two Mab react the aritigen encoded B-G sub- region. It was carried out to purify the chicken B-F antigen by affinity chromatography coupled with anti-B-F Mab. B-F antigen was analyzed by SDS-PAGE, and molecular weight was about 50KDa.
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Research Products
(4 results)