铜绿假单胞菌重组Bb-OprF疫苗诱导小鼠细胞免疫应答的研究

目的研究铜绿假单胞菌重组双歧杆菌(rBb)-OprF疫苗免疫小鼠,PAO1株攻击后产生的细胞免疫应答。方法将rBb-OprF疫苗分别采用皮下注射、肌肉注射、鼻腔黏膜和口服灌胃4种途径免疫Balb/c小鼠,免疫后8周用5×106CFU的PAO1株攻击,攻击1周后杀鼠取脾,MTT法检测特异性脾淋巴细胞增殖,流式细胞术检测脾CD4+T和CD8+T细胞比率,用ELISA法检测脾细胞培养上清IFN-γ、IL-12、TNF-α和IL-10的水平。结果脾T淋巴细胞增殖明显;脾细胞CD4+和CD8+T细胞亚群显著增加;脾细胞培养上清IFN-γ、IL-12、TNF-α和IL-10的水平显著升高,其中IFN-γ最为显著。结论铜绿假单胞菌rBb-OprF疫苗可诱导小鼠产生混合型的Th1和Th2免疫应答。

Cellullar immunologic response in mice induced by recombinant Bb-OprF vaccine of Pseudomonas aeruginosa against challenge of PAO1 strain

To observe changes of cellular immunity in Balb/c mice immunized with recombinant Bifidobacteria(rBb)-OprF vaccine of Pseudomonas aeruginosa and then challenged with PAO1,Balb/c mice were immunized with the vaccine subcutaneously,intramuscularly,intranasally and orally.Then the mice were challenged with 5.0×106 CFU PAO1 strain at 8th week after the vaccination,and sacrificed on 9th week.Methyltetrazolium(MTT) assay indicated that the proliferation of splenocytes was significantly increased;flow cytometry(FCM) showed that the number of CD4+ and CD8+ subsets in 4 immunization groups were more than that of control;ELISA demonstrated that the levels of IFN-γ,IL-12,TNF-α and IL-10 in the culture supernatants were markedly elevated.In conclusion,we presume that mixed Th1/Th2 type immunologic responses may be induced in mice immunized with the recombinant Bb-OprF vaccine of Pseudomonas aeruginosa.