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构建婴儿双歧杆菌口服活疫苗,表达产毒性大肠杆菌主要菌毛亚单位(CfaB)和不耐热肠毒素 B 亚单位(LTB)的抗原。

Construction of Bifidobacterium infantis as a live oral vaccine that expresses antigens of the major fimbrial subunit (CfaB) and the B subunit of heat-labile enterotoxin (LTB) from enterotoxigenic Escherichia coli.

机构信息

Key Laboratory of Biochemistry and Molecular Biology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.

出版信息

Microbiology (Reading). 2012 Feb;158(Pt 2):498-504. doi: 10.1099/mic.0.049932-0. Epub 2011 Nov 3.

Abstract

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.

摘要

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。

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