抗大肠杆菌_(506)株36kD蛋白单抗的制备及其在人精子上的免疫定位  

Production of Murine Monoclonal Antibodies (McAbs) Reactive to a 36kD Protein of Escherichia coli506 Strain and Immunological Localization of Cross-antigens in Human Sperm

作　　者：蒋玖[1] 陆德源[1] 曾瑞云 王红 徐奔 

机构地区：[1]上海第二医科大学微生物学教研室,上海200025 [2]上海市医学检验重点实验室,上海200023

出　　处：《上海免疫学杂志》1998年第3期147-149,共3页Shanghai Journal of Immunology

基　　金：上海市高教局资助项目(294-02)

摘　　要：用纯化的３６Ｄ蛋白免疫的Ｂａｌｂ／ｃ小鼠。取免疫脾细胞与小鼠骨髓瘤细胞ＳＰ２／０以ＰＥＧ进行细胞融合，融合率为８６％，采用ＥＬＩＳＡ筛选出杂交瘤细胞阳性孔。经过４－７次有限稀释克隆化，共获得７株能稳定分泌３６ＫＤ相应ＭｃＡｂ的杂交瘤细胞。Balb/c mice were immunized by intraperitoneal injection of purified 36kD protein. Spleen cells from one immunized mice were harvested and fused with myeloma line SP2/0 in the presence of polyethylene glycol. The resulting hybrid cells were screened according to their ability to bind to 36kD protein in plate ELISA. Hybrids secreting reactive antibodies were subcloned 4 to 7 times by limiting-dilution cloning and 7 hybrids were selected. Hybrid cells were grown as ascites in the peritoneal cavity of silica H primed Balb/c mice. McAbs were purified from ascitic fluid. It was showed that all the 7 clones were of the IgGl isotype and the liters were between 1 : 5×103- 1 : 1 ×105. Immunological localization of the cross-reactive antigens on human sperm with the 36kD protein was tested by means of indirect immunofluorescence. It was found that EC2 could react with the head and neck parts of human sperm. These results expand our understanding on the machanisms of male infertility induced by bacterial infection and have a great significance in preparing a potential effective immuno-contraceptive vaccine from bacteria in the near future.

关 键 词：人精子 大肠杆菌 共同抗原 单克隆抗体 免疫定位 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]