肺炎链球菌表面蛋白A多价DNA疫苗交叉免疫保护作用评价.docVIP

肺炎链球菌表面蛋白A多价DNA疫苗交叉免疫保护作用评价 张 巧，林科雄，姚 伟，李 琦，钱桂生，王长征，马千里 （400037重庆，第三军医大学新桥医院全军呼吸内科研究所全军呼吸病研究重点实验室观察免疫小鼠鼻咽部肺炎链球菌携带和腹腔攻击小鼠21生存情况携带肺炎链球菌菌落计数P＞0.05）明显小鼠P0.01）肺炎链球菌腹腔攻击中位生存时间P＞0.05）明显长于小鼠P0.01）。 结论R392233; R392. 7; R563. 1 [文献标志码] A Evaluation of Cross- protection of Streptococcus pneumoniae DNA vaccine with pspA gene against Streptococcus pneumoniae with different PspA Family clades Zhang Qiao , Lin Kexiong, Yao Wei, Li Qi, Qian Guisheng,Wang Changzheng, Ma Qianli ( Department of Respiratory Diseases, Xinqiao Hospital, Third Military Medical University. Chongqing, 400037, China）[Abstract] Objective To evaluate the ability of Streptococcus pneumoniae multivalent DNA vaccine with dominant pneumococcal surface protein A（pspA） clades genes against Streptococcus pneumoniae with different PspA family clades. Methodsolymerase chain reaction (PCR) amplified the PspA -Fam1-Clade1 and PspA-Fam2-Clade3 fragments from the recombinant T vectors. Which were antigens with a high protective antibody titers and strong cross-reaction against the other PspA family clades. And then they were cloned into the eukaryotic vector pcDNA3.1 (+) for the recombinant plasmid pcDNA3.1-Clade1 and pcDNA3.1-Clade3. Immunoblotting (Western-blot) and reverse transcription-polymerase chain reaction (RT-PCR) tests were made for detecting the expression of recombinant plasmids in mammalian cells BHK-21. Streptococcus pneumoniae PspA recombinant multivalent DNA vaccines (pcDNA3.1-Clade1 + pcDNA3.1-Clade3) were made to inject mice, and then the nasopharyngeal carriage of Streptococcus pneumoniae isolates with different PspA family clades in the immunized mice were evaluate, and the median survival time of the immunized mice attacked abdominal with Streptococcus pneumoniae isolates with different PspA family clades were counted after 21-day. Results The Streptococcus pneumoniae PspA recombinant multivalent DNA vaccines (pcDNA3.1-Clade1 + pcDNA3.1-Clade3) were successfully constructed with the domi