滋养层细胞促融合表位肽抗生育作用的初步研究  

Contraceptive effect of fusogenic trophoblast surface epitopes

作　　者：张英晨[1] 方针强[1] 叶钢[1] 

机构地区：[1]第三军医大学新桥医院泌尿外科全军肾脏病中心,重庆400037

出　　处：《第三军医大学学报》2007年第12期1218-1220,共3页Journal of Third Military Medical University

基　　金：重庆市计生委科技攻关项目(2004C062)~~

摘　　要：目的合成多价抗原肽,观察其体液免疫效果,并初步评估其抗生育潜力。方法选择人滋养层细胞促融合表位模拟肽与一种通用辅助T细胞表位(PADRE)作为骨架,合成多价抗原肽(MAP)结构的免疫原,与等量弗氏佐剂混悬后免疫雌性C57BL/6小鼠,观察体液免疫反应的特点及抗血清干扰滋养细胞融合的效果。结果在431A固相自动肽合成仪上成功地合成了MAP多肽,并经HPLC纯化、质谱鉴定证实其纯度达95%以上;其在小鼠血清中诱导出的特异性抗体的滴度最高达1:1024,且可以特异识别滋养层细胞相关抗原表位;在1:10稀释的抗血清存在时,forskolin诱导的人绒癌细胞(BeWo)间融合显著减少(P<0·01)。结论由含有人滋养层细胞促融合表位模拟肽、具有特异性氨基酸序列的新抗原所制备的抗血清可识别滋养层细胞相关天然抗原表位,并在小鼠体内激发出较理想的特异性体液免疫,同时,抗血清对BeWo细胞间融合具有一定抑制作用。Objective To synthesize a multiple antigenic peptide （MAP）, observe its humoral immune response and evaluate its antifertility potential. Methods Based on fusogenic epitope mimic peptide and synthetic nonnatural Pan DR T Helper Epitope （PADRE）, MAP antigenic peptides were designed and synthesized. After suspending with the equivalent Freund＇ s adjuvant, the antigens were used to immunize the female C57BL/6 mice to evalutate their humoral immune response in animals and efficiency of specific antisera on inhibiting syncytial trophoblast fusion. Results The MAP peptide was successfully synthesized. After purified with HPLC, its purity reached 95%. After immunization, the highest level of specific IgG in mice serum was 1 ： 1 024, and the corresponding antigen of human trophoblastic cell could be identified in the antisera. In presence of mice antiserum by 1 ： 10 dilution, forskolin-induced intercellular fusion of BeWo choriocarcinoma cells decreased remarkably （P 〈0. 01 ）. Conclusion The antibodies from the trophoblast fusogenic epitope mimic peptide with specific anmino acid sequence can identify the original antigens, and stimulate powerful special humoral immunity in mice, and inhibit trophoblast cell fusion in presence of the antisera. It provides an experimental basis for contraceptive vaccine in trophoblast cell antigen study.

关 键 词：滋养层细胞 免疫避孕 表位模拟肽 细胞间融合 

分 类 号：R169.4[医药卫生—公共卫生与预防医学] R341.6