布鲁杆菌基因工程疫苗免疫保护效率的初步观察

目的比较布鲁杆菌6种抗原表位所获得的基因工程疫苗的免疫保护效率。方法布鲁杆菌核糖体蛋白L7/L12、胞质蛋白P39、细胞表面蛋白BCSP31、二氧四氢喋啶合成酶BLS、16.5×103的外膜蛋白PAL和翻译起始因子IF3的基因片段与真核表达载体pcDNA3.1( )构建的核酸疫苗及上述6条片段转入pET32a( )后诱导表达的的重组蛋白免疫小鼠,3次免疫后进行攻毒实验,对其免疫保护效率进行初步观察。结果L7/L12和BLS的重组蛋白疫苗和核酸疫苗都产生了非常显著的保护作用,P39的核酸疫苗、IF3和BCSP31的重组蛋白疫苗也产生了非常显著的保护作用,其中L7/L12核酸疫苗的保护效率最高。结论初步认为布鲁杆菌的优势抗原表位所获得的基因工程疫苗可产生一定的抗布鲁杆菌作用,可作为未来布鲁杆菌新型疫苗的候选者,有进一步研究的价值。

Observation on protective effect of engineering vaccine against Brucella challenged by/Brucella abortus 544A on BALB/c Mice

Objective To compare the protection effects of six kinds of engineering vaccines against Brucella challenged by Brucella abortus 544A on BALB/c Mice. Methods Gene sequences of L7/L12, P39, BCSP31, BLS, pal and IF3 were cloned into the eurokaryotic expressing plasmid pcDNA3.1 ( ) separately. Mice were immunized with these recombinant plasmids via an intramuscularly route. These six gene sequences were also cloned into the prokaryotic expressing plasmid pET32a( ) separately, followed by the transfection of these recombinant plasmids into BL21. The fusion proteins of the six genes induced by IPTG were identified and purified. Mice were immunized with these recombinant proteins mixed with Freund's adjuvant. After three times of immunization, the titers of the antibody were evaluated by ELISA on the 60 day and the mice were challenged with Brucella abortus 544A. Brucella CFU at spleen was counted 20 days after challenge. Results Both kinds of vaccines of L7/L12 and BLS had a significant degree of protection, so did pcDNA3.1-P39, recombinant proteins of IF3 and BCSP31, among which, pcDNA3.1-L7/L12 was the most potent. Conclusions The engineering vaccines carrying antigen of brucella have a protective effect against brucella, and can be used as a candidate vaccine for the control of brucellosis.