Studies on the sub-region of chicken major histocompatibility B locus by using monoclonal antibodies.

鸡主要组织相容性复合体编码3类抗原：B-F(I类)、B-L(II类)和B-G(IV类)，表达于红细胞和红系前体细胞。对BALB/c小鼠进行腹腔免疫。分别于第0天和第14天注射2×10~(-7)红细胞、外周血淋巴细胞和脾淋巴细胞，末次静脉注射。用B^9B^9和B^&lt；11&gt；B^&lt；11&gt；红细胞免疫的小鼠脾细胞(融合1和融合2)、脾淋巴细胞(融合3-6)和外周血淋巴细胞(融合7和融合8)进行8次融合。用流式细胞仪(FCM)和血清学方法检测了6株单抗的特性。其中4例红细胞和外周血淋巴细胞均呈阳性反应。这一结果提示这四株单抗具有B-F亚区的特异性。6株单抗中有2株仅与红细胞反应。血清学结果还表明，这两株单抗均与编码B-G亚区的抗原发生反应，采用亲和层析与抗B-F单抗偶联的方法纯化鸡B-F抗原。经SDS-PAGE分析，B-F抗原的相对分子质量约为50 KDa。

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.