日本血吸虫童虫保护性单克隆抗体的建株和鉴定

PRODUCTION AND CHARACTERIZATION OF A MURINE PROTECTIVE MONOCLONAL ANTIBODY AGAINST SCHISTOSOMA JAPONICUM SCHISTOSOMULA

本研究获8株抗日本血吸虫(中国大陆株)童虫的单克隆抗体,从中确定一株具有体内保护性功能的单克隆抗体(编号为N15D9),其保护率范围为14-39％。经间接免疫荧光法和ELISA试验检测,N15D9能与尾蚴、3h龄机械转化童虫和5d龄肺童虫的表膜抗原反应。经3h龄机械童虫可溶性抗原的免疫印迹法分析,N15D9能识别132、96和10kDa抗原分子。

Eight murine monoclonal antibodies against surface determinants of Schistosoma japoni-cum (Chinese mainland strain) schistosomula were generated,of which only one monoclonal IgM antibody (N15D9) gave protection at level ranging from 14 to 39% in experiments of passive transfer or inhibition of infectivity while the others did not exhibit significant levels of passive protection.Further characterization of N15D9 antigen specificity showed that 96 and 14 kDa antigen molecules in cercaria,and 132 and 10 kDa in schistosomula could be recognized by N15D9 in Western blot assay.Furthermore,the 96 and 132 kDa molecules could also be recognized by pooled infected human sera while 14 kDa and 10 kDa only by sera from mice vaccinated with 3-hour schistosomula.The molecules recognizable by N15D9 were surface epitopes repeatedly expressed on cercaria,in vitro 3-hour mechanically transformed schistosomula and 5 day lung-stage schistosomula,as demonstrated by indirect immuno-fluorecence surface binding assay.