紫外线致弱尾蚴免疫兔血清筛选日本血吸虫童虫cDNA文库

目的筛选紫外线致弱尾蚴中起免疫保护作用的抗原分子,分析其主要表位,为血吸虫病疫苗研究提供新的候选抗原。方法以紫外线照射致弱尾蚴免疫兔血清筛选日本血吸虫童虫cDNA文库,对阳性克隆插入片段的核苷酸进行序列分析,并利用软件Antheprot对所筛基因编码的抗原分子进行表位预测。结果经三轮筛选,共获20个阳性克隆,序列同源性分析表明其中有5种已知基因(GST、副肌球蛋白、肌球蛋白、钙离子激活蛋白和3-磷酸甘油醛脱氢酶),6种未知基因,软件分析显示不同抗原分子分别具有多个不同的抗原表位。结论获得的阳性克隆插入基因片段可能为潜在的日本血吸虫病疫苗分子,可能是多抗原、多表位在照射致弱尾蚴中起免疫保护作用。

Screening of the cDNA library of Schistosomula japonicum by sera from rabbits immunized with ultaviolet-attenuated cercariae

To find out the major antigenic molecules of the ultraviolet-attenuated cercarise and to analyze their chief epitopes that may be used as the candidate antigens for the development of vaccines for schistosomiasis,the cDNA library of Schistosomula japonicum was screened with rabbit antisera against ultraviolet-attenuated cercariae,and the nucleotide sequence analysis of the insertion fragment of the positive clones was performed.The sequences were compared with all the sequences in Genbank by interent,and the epitopes of the screened genes encoding antigens were analyzed by Antheprot software. After three rounds of screening,20 positve clones were obtained,in which 5 known genes(genes coding for GST,paramyosin,myosin,calpain and SG3PDH) were proved via homologous comparison,and 6 unknown genes were found.Multiple antigenic epitopes could be detected on different antigenic molecules as revealed by software analysis.Thus,insertion gene fragment of the positive clones obtained may be the potential candidate antigen for the development of vaccines for schistosomiasis in the future.