LTB的表达及其粘膜免疫佐剂活性分析  

Expression and Analysis of Mucosal Immunoadjuvant Activity of LTB

作　　者：张国广[1,2] 罗茂春[3] 董艳美[1] 陈亮[1] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005 [2]漳州师范学院生物科学与技术系,福建漳州363000 [3]龙岩学院生命科学学院,福建龙岩364012

出　　处：《厦门大学学报（自然科学版）》2012年第5期918-922,共5页Journal of Xiamen University：Natural Science

基　　金：福建省科技厅重点项目(2009N0051);福建省自然科学基金项目(2010J01240);厦门市科技计划项目(3502Z20103007)

摘　　要：粘膜免疫是预防一些通过吸附粘膜组织侵入机体病原的很好措施,但粘膜免疫通常需要粘膜免疫佐剂,大肠杆菌(Escherichia coli)不耐热肠毒素的B亚基具有很好的粘膜免疫佐剂活性.本研究从大肠杆菌195菌株中扩增出LT(heat-labile enterotoxin)基因,测序后将其B亚基基因与pET32a连接构建了重组质粒pET32a-LTB,SDS-PAGE显示LTB(heat-labile enterotoxin B subunit)在原核细胞中得到表达,Western blotting和人神经节苷脂结合酶联免疫吸附试验(GM1-ELISA)结果表明,重组LTB具有抗原性和GM1结合活性.将其与禽流感M2e表位融合蛋白M2eHBc+混合通过滴鼻途径免疫小鼠,抗体检测结果表明,所表达的LTB能很好地提高共免疫抗原的粘膜和系统免疫应答.Mucosal immunity is a good measure for preventing pathogenic agents that occur at or emanate from mucosal surfaces invaded the body,but mucosal immunity usually need to mucosal immunoadjuvant. Escherichia coli heat-labile enterotoxin B subunit （LTB） has a good mucosal immunoadjuvant activity. LT gene was amplified by polymerase chain reaction from an enterotoxigenic E. coli 195 strain in this study. Prokaryotic expression vector pET32a-LTB was constructed by LTB subunit gene and pET32a connec tion after LT gene was sequenced. SDS-PAGE displayed that LTB subunit are expressed in E. coli cells and recombinant LTB inheri- ted the antigenicity and GMl-binding activity of cholerae enterotoxin by Western blotting and GM1-ELISA assay. The mice immuni zation with mixing of recombinant LTB and avian influenza virus M2e epitope fusion protein M2eHBc ＋by intranasal dropping way revealed that LTB can significantly enhance the mucosal and systemic immune responses of mice to co- administered antigen.

关 键 词：LTB表达 GM1-ELISA 粘膜免疫佐剂活性 

分 类 号：Q786[生物学—分子生物学]