PspA免疫小鼠抗肺炎链球菌侵袭性感染研究  

Protective effect of PspA against invasive infection of Streptococcus pneumoniae in mice

作　　者：胥文春[1] 曹炬[1] 许颂霄[1] 罗进勇[1] 朱旦[1] 尹一兵[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学省部共建教育部重点实验室,重庆400016

出　　处：《第四军医大学学报》2007年第9期783-786,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30471873)

摘　　要：目的:获取原核表达的肺炎链球菌PspA重组蛋白并研究其作为疫苗的价值.方法:分离培养肺炎链球菌TIGR4,获取其染色体DNA,采用基因体外重组法将编码PspA抗原表位在内的部分序列克隆到原核表达载体PET-32(a)内,酶切及测序鉴定重组质粒转化到大肠杆菌BL21(DE3)中,经IPTG诱导,将获得的重组蛋白用Western Blot鉴定,镍柱纯化并透析除盐.用重组蛋白免疫动物,观察PspA抗体对肺炎链球菌感染小鼠的保护作用.结果:DNA序列与Gen-Bank中的数据相符,所表达纯化的蛋白经Western Blot证实为PspA,其抗体在肺炎链球菌感染中对小鼠有保护作用.结论:重组PspA刺激产生的抗体能够有效抵抗肺炎链球菌TIGR4侵袭性感染,该重组蛋白可作为肺炎链球菌多肽联合疫苗的组成部分之一.AIM： To obtain purified PspA by prokaryotic expression system and to investigate the effect of PspA antibody on invasive infection of Streptococcus pneumoniae. METHODS： Template DNA was isolated from cultured TIGR4. By gene recom- bination technology in vitro, the sequence encoding PspA antigen epitope was cloned into PET-32 （a） expression vector. After being confirmed by sequencing, expressed antigen protein was identified by Western Blot and purified through Ni^2＋ affinity chromatography and processed by dialysis and preserved by freezing and drying. After pneumococcal challenge in actively and passively immunized mice, we compared their median survival times by Mann-Whitney U test. RESULTS： The DNA sequence of the cloned PspA gene was consistent with GenBank data, and PspA fusion protein was proved by Western Blot. PspA can exert a protection against inva- sive pneumococcal infection. CONCLUSION： A highly expressed recombinant PspA protein has been successfully obtained and proved to exert the protection against invasive pneumococcal infection, suggesting PsoA can be a oromising candidate vaccine.

关 键 词：肺炎链球茵 肺炎链球茵表面蛋白A 疫苗 

分 类 号：R392.1[医药卫生—免疫学]