期刊
FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY
卷 57, 期 1, 页码 69-79出版社
WILEY
DOI: 10.1111/j.1574-695X.2009.00580.x
关键词
Vibrio cholerae; bioluminescent imaging; promoter; vector vaccine; in vivo expression
资金
- 'Research Training in Gastroenterology' [T32 DK067872]
- [R0I AI19716]
Live, attenuated Vibrio cholerae vaccines can induce potent immune responses after only a single oral dose. The strategy of harnessing these strains to present antigens from heterologous pathogens to the mucosal immune system shows great promise. To fully realize this possibility, V. cholerae strains must be created that stably express antigens in vivo in sufficient quantity to generate an immune response. In vivo-induced promoters have been shown to increase the stability and immunogenicity of foreign antigens expressed from multicopy plasmids. We report the construction of a series of genetically stabilized plasmids expressing luciferase as a heterologous protein from the following in vivo-induced promoters: V. cholerae P-argC, P-fhuC and P-vca1008, and Salmonella enterica serovar Typhi P-ompC. We demonstrate that several of these expression plasmids meet two critical criteria for V. cholerae live vector vaccine studies. First, the plasmids are highly stable in the V. cholerae vaccine strain CVD 103-HgR at low copy number, in the absence of selective pressure. Second, real-time bioluminescent imaging (BLI) demonstrates inducible in vivo expression of the promoters in the suckling mouse model of V. cholerae colonization. Moreover, the use of BLI allows for direct quantitative comparison of in vivo expression from four different promoters at various time points.
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