中国兽医科学2018，Vol.48Issue(5)：545-551,7.DOI:10.16656/j.issn.1673-4696.2018.0097
猪免疫调控分子B7-H4的单克隆抗体制备及其在免疫细胞中的表达分析
Production of monoclonal antibodies against porcine immune regulatory molecule B7-H4 and analysis of expression of B7-H4 in immune cells
摘要
Abstract
B7-H4 is one member of B7 family. Many researches in mice and humans show that B7-H4 plays an important regulatory role on the function of T cells. However,there is no report on the function of B7-H4 in porcine immune system so far. In this research,the gene fragment of porcine B7-H4(pB7-H4)was amplified by PCR and cloned into prokaryotic expression vector pET-32a(+) to obtain pET-32a-pB7-H4. SP2/0 myeloma cells were fused with spleen cells from the BALB/c mice immunized with purified recombinant pB7-H4 expressed in E. coli. One hybridoma cell line,named as 3C8,stably secreting MAb against pB7-H4 protein was identified by indirect ELISA. The MAb is IgGl subtype. Flow cytometic analysis showed the MAb can specifically bind with the pB7-H4 temporally expressed on a cell line. It was used to analyze the pB7-H4 expression on the porcine immune cells. The results indicated that pB7-H4 molecule is highly expressed on macrophages in both porcine pulmonary alveoli and peripheral blood mononuclear cell(PBMC), but almost not on T lymphocytes. In conclusion,the successful production of MAb specific for pB7-H4 provides a useful tool for further research on pB7-H4 immune functions.关键词
猪B7-H4/原核表达/单克隆抗体/免疫细胞/流式细胞检测Key words
porcine B7-H4/prokaryotic expression/monoclonal antibody/immune cells/flow cytometry analysis分类
农业科技引用本文复制引用
郭杰,刘路,朱可蒙,王宁,辛九庆,刘恒贵..猪免疫调控分子B7-H4的单克隆抗体制备及其在免疫细胞中的表达分析[J].中国兽医科学,2018,48(5):545-551,7.基金项目
中央级公益性科研院所基本科研业务费专项(1610302016005 （）
1610302017014) （）
