运用逆向疫苗学对沙眼衣原体疫苗候选蛋白的初步筛选  

Reverse vaccinology for preliminarily screening Chlamydia trachomatis vaccine candidate proteins

作　　者：王伟[1] 张强[1] 王海梅[1] 郭普[1] 钟政荣[1] 

机构地区：[1]蚌埠医学院第一附属医院检验科,蚌埠233004

出　　处：《山西医科大学学报》2015年第10期986-990,共5页Journal of Shanxi Medical University

基　　金：安徽省高校省级自然科学基金资助项目(KJ2012Z247)

摘　　要：目的探讨运用逆向疫苗学理论,筛选沙眼衣原体疫苗候选蛋白的可行性。方法通过对沙眼衣原体测序基因组中890个基因进行生物信息学分析,预测编码表面暴露蛋白的基因,进一步利用实时定量PCR、蛋白质表达纯化、免疫动物以及特异性抗体效价的测定,验证候选蛋白的免疫原性。结果经过生物信息学分析,结构预测及亚细胞定位,共得到126个符合要求的蛋白。排除与人体可能存在同源的蛋白后,共得到73个候选蛋白。利用实时定量PCR分析其中30个蛋白编码的基因,挑选其中表达量比较高的4个进行免疫效果的评价。实验证实这4个基因编码的蛋白均可以产生保护性抗体。结论本研究证实逆向疫苗学理论在沙眼衣原体疫苗的抗原筛选工作中是可行的,为下一步寻找有效的候选疫苗抗原打下了良好的基础。Objective To investigate the feasibility of screening candidate proteins of Chlamydia pneumoniae vaccine based on the theory of reverse vaccine. Methods Bioinformation of 890 genes in the whole genome of Chlamydia pneumonia was analyzed to predict genes coding surface exposed proteins. The immunogenicity of candidate proteins was verified by real time quantitative PCR,proteins expression and purification,determination of antibody titers and specificity. Results Based on bioinformatics analysis,structure prediction and subcellular localization,126 proteins were selected. After ruling out potential humanity homologous proteins,73 candidate proteins were obtained. Among them,30 hypothetical proteins were analyzed with real time quantitative PCR,the immune effects of 4 higher expression proteins were assessed with animal experiments,and the ultimate results showed that 4 proteins coded with high-expressed genes induced the production of protective antibodies. Conclusion The research confirms the feasibility of reverse vaccinology theory for screening Chlamydia vaccine candidate antigens and contributes to the further effort for acquiring effective candidate antigen.

关 键 词：逆向疫苗学 沙眼衣原体 候选蛋白 

分 类 号：R374[医药卫生—病原生物学]