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

利用单克隆抗体研究鸡主要组织相容性B位点亚区。

基本信息

批准号：
63560261
负责人：
YAMAMOTO Yoshio
金额：
$ 1.02万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63560261/
关键词：
Chicken MHC Monoclonal antibody T lymphocyte Blood group モノクローナル抗体主要組織適合性遺伝子

项目摘要

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.

鸡的主要组织相容性复合体（MHC）编码三类抗原：B-F（I类）、B-L（II类）和B-G（IV类），后者表达于红细胞和红系前体细胞中，用已知B基因型的鸡作为细胞供体免疫小鼠。在第0天和第14天分别用2 × 10^7红细胞、外周淋巴细胞和脾淋巴细胞腹腔注射免疫BALB B/c小鼠，并在第21天接受最后一次静脉注射5 × 10^6细胞。用来自用B^9 B^9和B^ B^红细胞免疫的小鼠的脾细胞进行8次融合<11><11>（融合体1和2）、脾淋巴细胞（融合体3-6）和外周淋巴细胞（融合体7和8）。用流式细胞仪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。这一结果表明，这四个单克隆抗体具有B-F亚区的特异性。6株单抗中有2株仅与红细胞反应。血清学结果表明，这两株单抗均与鸡B-F抗原编码区的B-G亚区反应。经SDS-PAGE分析，B-F抗原分子量约为50 KDa。