Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Anzio Rd, Observatory 7925, Cape Town, South Africa.
Gut Pathog. 2009 Apr 29;1(1):9. doi: 10.1186/1757-4749-1-9.
The induction of antigen-specific CD8+ T cell cytokine responses against an attenuated, oral recombinant Salmonella enterica serovar Typhimurium vaccine expressing a green fluorescent protein (GFP) model antigen was investigated. A GFP expression plasmid was constructed in which the gfp gene was fused in-frame with the 5' domain of the Escherichia coli beta-galactosidase alpha-gene fragment with expression under the lac promoter. Groups of mice were orally immunized three times with the bacteria and systemic CD8+ T cell cytokine responses were evaluated.
High level of the GFP model antigen was expressed by the recombinant Salmonella vaccine vector. Systemic GFP-specific CD8+ T cell cytokine (IFN-gamma and IL-4) immune responses were detected after mice were orally vaccinated with the bacteria. It was shown that 226 net IFN-gamma and 132 net IL-4 GFP-specific SFUs/10e6 splenocytes were formed in an ELISPOT assay. The level of IFN-gamma produced by GFP peptide-stimulated cells was 65.2-fold above background (p < 0.05). The level of IL-4 produced by the cells was 10.4-fold above background (p < 0.05).
These results suggested that a high expressing recombinant Salmonella vaccine given orally to mice would elicit antigen-specific CD8+ T cell responses in the spleen. Salmonella bacteria may, therefore, be used as potential mucosal vaccine vectors.
本研究旨在探讨减毒口服重组沙门氏菌 Typhimurium 疫苗诱导针对绿色荧光蛋白(GFP)模型抗原的特异性 CD8+T 细胞细胞因子应答的情况。构建了一个 GFP 表达质粒,该质粒将 gfp 基因与大肠杆菌β-半乳糖苷酶α-基因片段的 5' 结构域融合,并在 lac 启动子的控制下表达。将小鼠分组口服免疫三次该细菌,并评估系统 CD8+T 细胞细胞因子(IFN-γ和 IL-4)应答。
重组沙门氏菌疫苗载体高水平表达 GFP 模型抗原。用细菌口服接种后,检测到系统性 GFP 特异性 CD8+T 细胞细胞因子(IFN-γ和 IL-4)免疫应答。ELISPOT 检测结果显示,10e6 个脾细胞中有 226 个 GFP 特异性 IFN-γ和 132 个 GFP 特异性 IL-4 SFUs。GFP 肽刺激细胞产生的 IFN-γ水平比背景高 65.2 倍(p < 0.05)。细胞产生的 IL-4 水平比背景高 10.4 倍(p < 0.05)。
这些结果表明,口服给予小鼠的高表达重组沙门氏菌疫苗可在脾脏中引发针对抗原的特异性 CD8+T 细胞应答。因此,沙门氏菌可能被用作潜在的黏膜疫苗载体。