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首页> 外文期刊>Vaccine >Immunization with recombinant Lactobacillus casei strains producing K99, K88 fimbrial protein protects mice against enterotoxigenic Escherichia coli
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Immunization with recombinant Lactobacillus casei strains producing K99, K88 fimbrial protein protects mice against enterotoxigenic Escherichia coli

机译：用产生K99，K88纤维蛋白的重组干酪乳杆菌菌株免疫可保护小鼠免受产肠毒素的大肠杆菌的侵害

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To exploit a safe and effective vaccine for the prevention against K99 or K88 infections of enterotoxigenic Escherichia coil (ETEC), we have developed a mucosal delivery vehicle based on Lactobacillus casei CICC 6105 using poly-gamma-glutamate synthetase A (PgsA) as an anchoring matrix. To evaluate the immunization effect of the recombinant strains (harboring plasmids pLA-K99-K88-LTB, pLA-K99, and pLA-K88), anti-ETEC K99 or K88 antibody responses, T-cell proliferation, and cytokines by intracellular staining (ICS) were investigated after specific pathogen-free (SPF) C57BL/6 mice orally inoculated with these recombinant strains. After oral vaccination into C57BL/6 mice, all recombinant strains were proved to be immunogenic and able to elicit high levels of mucosal immunoglobulin A (IgA) titers in bronchoalveolar lavage fluids, intestinal fluids and prominent systemic immunoglobulin G and IgG subclasses (IgG1, IgG2b, and IgG2a) responses in sera. Using the T-cell proliferation assay, the stimulation index (SI) of groups immunized with pLA-K99/L. casei and pLA-K88/L. casei reached to 2.73 and 2.64, respectively, versus 2.56 in a group immunized with pLA-K99-K88-LTB/1.. casei. A detailed analysis of the cell-mediated immune responses by ICS showed the number of specific CD8(+) T cells expressing cytokines (IFN-gamma, INF-alpha, and IL-2) and granule-associated proteins (CD107a) was higher than that of specific CD4(+) T cells secreted by immune spleen cells upon restimulation in vitro with peptides. Next, the results showed that DCs activated in vitro with recombinant L. casei enhance specific T-cell proliferation and promote T cells to produce both Th1 and Th2 cytokines. More than 80% of the vaccinated mice were protected after challenge with a lethal dose of standard strains C83912 and C83902. These results demonstrate that recombinant L casei can induce specific humoral and mucosal antibodies and cellular immune response against protective antigens upon oral administration. (c) 2011 Elsevier Ltd
机译：为了开发一种安全有效的疫苗来预防肠毒素性大肠埃希氏大肠杆菌（ETEC）的K99或K88感染，我们开发了一种基于干酪乳杆菌CICC 6105的粘膜输运载体，使用聚γ-谷氨酸合成酶A（PgsA）锚定矩阵。为了评估重组菌株（携带质粒pLA-K99-K88-LTB，pLA-K99和pLA-K88的疫苗），抗ETEC K99或K88抗体的应答，T细胞增殖和细胞因子通过细胞内染色的免疫效果（在口服这些重组菌株的特定无病原体（SPF）C57BL / 6小鼠后，研究了ICS）。在对C57BL / 6小鼠进行口服疫苗接种后，所有重组株均被证明具有免疫原性，并能够在支气管肺泡灌洗液，肠液以及突出的全身性免疫球蛋白G和IgG亚类（IgG1，IgG2b）中引起高水平的粘膜免疫球蛋白A（IgA）滴度。和IgG2a）的血清反应。使用T细胞增殖测定法，用pLA-K99 / L免疫的组的刺激指数（SI）。酪蛋白和pLA-K88 / L。在用pLA-K99-K88-LTB / 1。casei进行免疫的组中，casei分别达到2.73和2.64，而2.56。 ICS对细胞介导的免疫反应的详细分析显示，表达细胞因子（IFN-γ，INF-α和IL-2）和颗粒相关蛋白（CD107a）的特定CD8（+）T细胞数量高于免疫脾细胞在体外用肽再刺激后分泌的特定CD4（+）T细胞的表达。接下来，结果显示，体外用重组干酪乳杆菌激活的DC增强了特异性T细胞增殖并促进T细胞产生Th1和Th2细胞因子。致死后，用致死剂量的标准菌株C83912和C83902保护了80％以上的接种小鼠。这些结果表明，重组L干酪素在口服时可以诱导特异性的体液和粘膜抗体以及针对保护性抗原的细胞免疫应答。 （c）2011爱思唯尔有限公司

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