一株与正常人细胞成分反应的巨细胞病毒单克隆抗体的鉴定  

PRODUCTION AND CHARACTERIZATION OF A MONOCLONAL ANTIBODY TO HUMAN CYTONEGALOVIRUS RECOGNIZING NORMAL CELL PROTEINS

作　　者：俞慕华[1] 张咏南[1] 龚蕴贞[1] 

机构地区：[1]安徽医科大学微生物教研室

出　　处：《中华微生物学和免疫学杂志》1994年第3期203-205,共3页Chinese Journal of Microbiology and Immunology

摘　　要：用ＨＣＭＶ－ＡＤ１６９株抗原建立了一株分泌ＨＣＭＶＭｃＡｂ的杂交瘤细胞株Ｎ４Ｅ６。经Ｗｅｓｔｅｒｎｂｌｏｔ分析发现ＭｃＡｂ与ＨＣＭＶ抗原和正常人成纤维细胞（ＨＥＦ）抗原均有多条免疫沉淀带出现：与ＨＣＭＶ－ＡＤ１６９株抗原反应的沉淀带为８９Ｋ，８７Ｋ，７３Ｋ和４２Ｋ；与Ｄａｖｉｓ株抗原反应的为８９Ｋ，８７Ｋ，４４Ｋ；与８１６地方株抗原反应的为８９Ｋ，８７Ｋ，７３Ｋ，４４Ｋ；与ＨＥＦ抗原反应的为８９Ｋ，８７Ｋ，４４Ｋ。表明Ｎ４Ｅ６除能识别ＨＣＭＶ－ＡＤ１６９株和８１６地方株特异成分７３Ｋ，外，尚能识别ＨＣＭＶ和ＨＥＦ共同成分８９Ｋ，８７Ｋ和４４Ｋ。One clone of hybridoma cells,N4E6,secreting McAb to HCMV was established.It did not react with otherherpes viruses including HSV 1,HSy 2 and VZV.By indirect immunofluorescence and immunoenzymatic staining,antigens could be detected with McAb N 4E 6 both in infected and uninfected human cells ,but not in cells of nonhu-man origin such as:chicken embryo fibroblasts and rabbit kidney cells. A ntigens were present at the membrane ofhuman cells and dispersed diffusely within the cytoplasm. By Western blot analysis,McAb N4E6 reacted with polypeptides 89K,87K and 44K from extracts of humanembryo fibroblasts(HEF),89K,87K,73Kand 42K from HEF infected strain HCMV-AD169,89K,87K and 44Kfrom strain Davis,89K.87K,73K and 44K from fresh isolated strain 816.It appeared that McAb N4E6 did not onlyrecognize protein 73K specifically in HCMV,but also recognize proteins 89K,87K,44K both in HCMV and HEP.

关 键 词：巨细胞病毒 单克隆抗体 免疫印迹 

分 类 号：R373[医药卫生—病原生物学]