一种白细胞共同抗原的鉴定及其对B细胞活化的影响

王运平　张淑珍　朱立平

　　中国图书分类号　R392.11
　　摘　要　目的：用单克隆抗体鉴定白细胞共同抗原并对其性质进行初步分析。方法：用B细胞株3D5免疫Balb/c小鼠得到单抗5H12，借助间接免疫荧光及流式细胞仪分析，检测5H12抗原在多种白细胞表面的表达情况，并从白细胞膜中提取5H12抗原，通过增殖试验、聚丙烯酰胺凝胶电泳及免疫印迹对其特性进行初步分析。结果：5H12抗原表达于多种白细胞表面，包括休止T细胞、休止B细胞、单核细胞、中性粒细胞、体外PHA激活的T细胞、体外anti-μ激活的B细胞、也表达在T细胞株(Jurkat、Peer)、B细胞株(3D5、Daudi、CESS、BJAB)、单核细胞株U937和髓细胞株K652表面。增殖试验表明，该抗原与相应单抗结合后导致B细胞活化抑制。对抗原的初步分析显示：5H12是一种单链蛋白质分子，分子量为22 kD。结论：5H12是一种单链膜蛋白分子，属于白细胞共同抗原，与B细胞活化有关。
　　关键词　白细胞共同抗原　5H12　分子量

Identification of leukocyte common antigen (LCA) 5H12 and discovery of its effect on B cell activation

WANG Yun-Ping,ZHANG Shu-Zhen,ZHU Li-Ping.
Department of Immunology,Binzhou Medical College,Binzhou 256603

　　Abstract　Objective:To identify leukocyte common antigen(LCA) with monoclonal antibody(McAb) and analyze the LCA.Methods:Immunized Balb/c mice with B cell line 3D5 and obtained McAb 5H12，then detected the expression of 5H12 antigen on various kinds of leukocytes and white cell lines by indirect immunofluorescence and flowcytometric analysis and finally isolated and purified 5H12 antigen from cell membrane and analyzed the 5H12 by proliferation test ,SDS-polyacrylamide gel electrophoresis(SDS-PAGE) and Western-blotting.Results:5H12was expressed on a panel of human leukocytes and white cell lines including resting T cells,resting B cells,monocytes,neutrophils,PHA-activated T cells,anti-μ-activated B cells,T cell lines(Jurkat,Peer),B cell lines(3D5,Daudi,CESS,BJAB),monocyte cell line U937 and myeloid cell line K652.Combination of 5H12 antigen with 5H12 McAb could induce inhibition of B cell activation in a dose-dependent manner and this suggested that 5H12 might play a role in the process that led to B cell activation and proliferation.SDS-PAGE and Western-blotting analysis indicated that 5H12 is a 22 kD one-peptide-chain protein.Conclusion:5H12 is a one-peptide-chain membrane protein molecule,it is a LCA and related to cell activation.
　　

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