分化抗原gp42在人免疫细胞的表达  

Expression of differentiation antigen gp42 on human immune competent cells

作　　者：汪燚[1] 刘音[1] 马凤蓉[1] 史耕先[1] 赵方萄[1] 朱立平[1] 

机构地区：[1]中国医学科学院基础医学研究所中国协和医科大学免疫学系,北京100005

出　　处：《中华微生物学和免疫学杂志》2003年第6期468-471,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家 973资助项目 ( 2 0 0 1CB5 10 0 0 4);国家自然科学基金资助项目 ( 3 0 0 70 70 4)

摘　　要：目的　明确gp4 2蛋白为分化抗原 ,检测它在人免疫细胞的表达。方法　用gp4 2蛋白免疫小鼠 ,制备抗gp4 2蛋白的单抗。用反义技术制备gp4 2蛋白表达抑制的 3D5细胞。用免疫荧光染色和Western印迹确定gp4 2单抗的特异性。用胞膜蛋白免疫沉淀和流式细胞术确定gp4 2蛋白为分化抗原。用双色免疫荧光染色流式细胞术检测gp4 2分化抗原在多种人免疫细胞的表达。结果　制备了抗gp4 2蛋白的特异性单抗。用gp4 2单抗作探针 ,从 3D5细胞膜免疫沉淀到相对分子质量 (Mr)为4 2 0 0 0的一条蛋白带 ,并在活 3D5细胞见阳性免疫荧光染色。gp4 2分化抗原在人外周围血CD19+ 、CD14 + 、CD4 + 、CD8+ 和CD5 6 + 细胞的表达率分别为 5 9.71%± 11.36 %、89.0 8%± 4 .87%、2 6 .0 3%±9.75 %、2 4 .2 4 %± 15 .2 9%和 2 4 .2 0 %± 15 .72 % ,在人免疫细胞株的表达率分别为 5 8.0 % (Nalm6 )、72 .7% (3D5 )、5 3.6 % (BJAB)、6 1.0 % (Daudi)、6 .9% (SKW6 )、6 .3% (Jurkat)、5 .8% (Peer)、2 1.3% (K5 6 2 )和 39.1% (U937)。结论　gp4 2蛋白为一个分化抗原 ,在人外周围血CD14 + 细胞的表达率最高 ,其次为CD19+ 细胞 ,CD4 + 、CD8+ 和CD5 6 + 细胞的表达率较低。Objective To confirm that gp42 protein is a differentiation antigen. To detect expression of differentiation antigen gp42 on human immune competent cells. Methods McAb gp42 was developed by immunization of mouse with gp42 protein. 3D5 cells with inhibition of gp42 protein expression was developed by antisense technique. The specificity of McAb gp42 was measured using Western blot and flowcytometry. Its nature of differentiation antigen was identified by immunoprecipitation test on intact cell and flowcytometric assay on live cell with McAb gp42 as a probe. Expression of gp42 differentiation antigen on human immune competent cells was assayed by means of two colour staining and flowcytometry. Results McAbs specifically to gp42 protein were developed. With McAb gp42 as a probe a protein band with molecular weight 19kDa was immunoprecipitated from intact 3D5 cell and positive staining was observed on live 3D5 cell. The percentage of gp42 differentiation antigen on CD14+, CD19+, CD4+, CD8+ and CD56+ cell from human peripheral mononuclear cells was 89.08%±4.87%, 59.71%±11.36%, 26.03%±9.75%, 24.24%±15.29% and 24.20%±15.72%, respectively. The percentage of 5C5 differentiation antigen on human immune cell lines was 58.0% (Nalm6), 72.7% (3D5), 53.6% (BJAB), 61.0% (Daudi), 6.9% (SKW6), 6.3% (Jurkat), 5.8%(Peer), 21.3% (K562) and 39.1% (U937). Conclusion gp42 protein is a differentiation antigen. Differentiation antigen gp42 is heavily expressed on CD14+ cells, at medium on CD19+ cells, and rather weak on CD4+, CD8+ and CD56+ cells.

关 键 词：分化抗原gp42 免疫细胞 单克隆抗体 免疫荧光染色法 WESTERN印迹 

分 类 号：R392[医药卫生—免疫学]