Key Laboratory of Biochemistry and Molecular Biology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.
Microbiology (Reading). 2012 Feb;158(Pt 2):498-504. doi: 10.1099/mic.0.049932-0. Epub 2011 Nov 3.
We sought to develop Bifidobacterium infantis (BI) as a vehicle for the expression of heterologous antigens. Two proteins of enterotoxigenic Escherichia coli (ETEC) were expressed in BI: CfaB, a major fimbrial subunit protein, and LTB, the B subunit of heat-labile enterotoxin. The expression of CfaB and LTB in BI was verified by electrophoretic analysis. Sprague-Dawley rats were then subjected to intragastric immunization with BI-CfaB and BI-LTB systems both separately and together. ELISA was used to characterize the serum and mucosal immune responses against ETEC antigens. The immunized rats were intraperitoneally challenged with wild-type ETEC H10407 to study the immune response in vivo. The serum titres of IgG and faecal IgA antibodies in the BI-CfaB plus BI-LTB mixed vaccination group were significantly greater than those in the other two groups, which were immunized with a single vaccine (P<0.05). However, no significant difference was seen between the two groups that received a single immunization. These results suggest that expressing CfaB and LTB in BI provides a probiotic system with immunogenic properties. Furthermore, the expression of LTB in BI preserved its mucosal adjuvant effect. So this study confirms that BI can be used as a novel oral vaccine expression system for a heterologous antigen and BI-LTB can provide mucosal adjuvant properties.
我们试图将婴儿双歧杆菌(BI)开发为表达异源抗原的载体。两种肠毒素性大肠杆菌(ETEC)的蛋白在 BI 中表达:CfaB,主要的菌毛亚基蛋白,和 LTB,不耐热肠毒素的 B 亚基。CfaB 和 LTB 在 BI 中的表达通过电泳分析得到验证。然后,用 BI-CfaB 和 BI-LTB 系统分别和同时对 Sprague-Dawley 大鼠进行胃内免疫。ELISA 用于表征针对 ETEC 抗原的血清和粘膜免疫反应。用野生型 ETEC H10407 对免疫大鼠进行腹腔内攻击,研究体内免疫反应。在 BI-CfaB 加 BI-LTB 混合疫苗组中,血清 IgG 和粪便 IgA 抗体的滴度明显高于其他两组(P<0.05),这两组分别用单一疫苗免疫。然而,接受单一免疫的两组之间没有观察到显著差异。这些结果表明,在 BI 中表达 CfaB 和 LTB 提供了具有免疫原性的益生菌系统。此外,LTB 在 BI 中的表达保留了其粘膜佐剂效应。因此,本研究证实 BI 可用于表达异源抗原的新型口服疫苗表达系统,并且 BI-LTB 可提供粘膜佐剂特性。