弧菌外膜蛋白OmpU的免疫交叉反应性和交叉保护性  

Immunological cross-reactivity and cross-protection of outer membrane protein Omp U among Vibrio species

作　　者：伦镜盛[1] 张设熙 董亚萍[1] 谢少凤[1] 麦亨 梅颖怡 胡忠[1] 

机构地区：[1]汕头大学生物系微生物实验室,广东汕头515063

出　　处：《微生物学报》2016年第5期867-879,共13页Acta Microbiologica Sinica

基　　金：科技部农业科技成果转化资金(2013GB2E000368);广东省科技计划(2012B020308008);汕头市科技计划(67)~~

摘　　要：【目的】通过对弧菌外膜蛋白Omp U的克隆、表达以及免疫学特性分析,明确外膜蛋白Omp U是否为弧菌的共同抗原,并具有免疫交叉反应性和交叉保护性。【方法】对弧菌外膜蛋白omp U基因进行克隆和生物信息学分析。分别制备副溶血弧菌、溶藻弧菌、创伤弧菌、拟态弧菌和霍乱弧菌的Omp U重组蛋白抗血清,对Omp U的免疫交叉反应特性以及抗原表位定位情况进行比较分析。以霍乱弧菌的Omp U重组蛋白免疫小鼠后,再以多种弧菌进行攻毒,分析其交叉免疫保护作用。【结果】外膜蛋白Omp U在弧菌种内和种间相似性分别为73.0%–100%和58.6%–89.0%,并至少存在9个保守的B细胞抗原表位。Omp U重组蛋白抗血清在弧菌种内和种间均产生显著的免疫交叉反应,识别弧菌中分子量35–40 k Da的同源蛋白。副溶血弧菌ATCC17802、创伤弧菌ATCC27562和拟态弧菌ATCC33653来源的Omp U重组蛋白抗体能识别供试菌株,提示这些菌株的Omp U抗原表位定位于细胞表面。Omp U重组蛋白对免疫后的小鼠具有交叉免疫保护作用,攻毒实验后小鼠相对存活率(RPS)为43.0%–100%。【结论】上述结果表明,外膜蛋白Omp U是弧菌中一种保守的共同抗原,具有免疫交叉保护性,可以作为弧菌广谱疫苗的候选抗原。[Objective] To study the immunological cross-reactivity and cross-protection characteristics of Omp U in Vibrio species. [Methods] The omp U genes from 10 Vibrio strains were cloned, sequenced, followed by bioinformatics analysis. Western blot and whole-cell ELISA assay were used respectively to determine immunological cross-reaction feature and subcellular location of Omp U with rabbit serum against recombinant Omp U from V.parahaemolyticus ATCC17802, V. alginolyticus ATCC33787, V. vulnificus ATCC27562, V. mimicus ATCC33653 and V. cholera Vb0. Finally, the cross-protective property of recombinant Omp U（V. cholera-derived） was evaluated through vaccination and subsequent challenge with heterogeneous virulent Vibrio strains in mice. [Results] The similarities of Omp U proteins of Vibrio ranged from 73.0 to 100% intra-species, and from 58.6 to 89.0% inter-species.Furthermore, homologous epitopes were found in Omp U and shared by different species of Vibrios. Western blot of rabbit serum against recombinant Omp U showed cross-recognition intra- and inter-species. Bands were observed ranging from 35 to 40 k Da. Whole-cell ELISA assay further confirmed that the antiserum of recombinant Omp U from V. parahaemolyticus ATCC17802, V. vulnifgicus ATCC27562 and V. mimicus ATCC33653 recognized the tested Vibrio species, implying that epitopes of Omp U were located on the cell surface. Recorded relative percent survival of the vaccinated group varied from 43.0 to 100%, showing that mice were protected from Vibrio infection after immunization with Omp U protein. [Conclusion] Omp U was a conserved antigen among tested Vibrio species and might be a universal vaccine candidate for the prevention of Vibriosis.

关 键 词：弧菌 外膜蛋白 OMP U 交叉反应性 交叉保护性 

分 类 号：Q939.91[生物学—微生物学]