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Homeostatic properties of Lactobacillus jensenii engineered as a live... » Isaúde
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BMC microbiology [electronic resource]
2013-01-09 02:07:34

Homeostatic properties of Lactobacillus jensenii engineered as a live vaginal anti-HIV microbicide

Descrição: Background:Vaginal probiotics are investigated as a binary strategy for prevention of bacterial vaginosis and HIV. We applied an innovative experimental model using primary and immortalized human cervical and vaginal epithelial cells to assess the functional properties of Lactobacillus jensenii, a predominant constituent of the healthy vaginal microbiome, engineered to express the HIV-1 entry inhibitor modified cyanovirin-N (mCV-N). In this model bacteria colonize the epithelial cells over a period of 24-72 h. Staurosporine and the Toll-like receptor 2/6 ligand macrophage-activating lipopeptide-2 (MALP-2) serve as positive controls for apoptosis and proinflammatory activation, respectively. In 24-hour intervals, the colonized epithelium is assessed microscopically, supernatants are collected for measurement of soluble immunoinflammatory mediators and production of CV-N, and cells are lysed for assessment of: 1) apoptosis by cleaved versus total caspase-3 assay; 2) NF-kappaB activation by a luciferase reporter assay; or 3) epithelia-associated colony forming units (CFU) in Brucella agar.Results:Wild type (WT) L. jensenii 1153 consistently colonized cervical and vaginal cells in the absence of epithelial damage and apoptosis. The bioengineered derivatives expressing mCV-N or control plasmids showed the same stable colonization pattern, which was reproducible between technologists and bacterial batches (CFU coefficient of variation

Identificador: doi:10.1186/1471-2180-13-4
Volume: 0
Página: 4 a

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