| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1989: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
|---|
| 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.
|
