Interaction of Vibrio cholerae O1 El Tor vesicles with target cells
DOI:
https://doi.org/10.17072/1994-9952-2026-2-179-190Keywords:
Vibrio cholerae, outer membrane vesicles, LPS, cholera toxin, adhesion to target cellsAbstract
Cultivation of the typical strain and genovariants of Vibrio cholerae O1 for 18 hours on Marten agar supplemented with 0.1% bile induced the formation of outer membrane vesicles (OMVs). TEM analysis showed that V. cholerae O1 El Tor vesicles in both liquid and solid media have an intact, continuous membrane. Visual inspection of the vesicle preparations revealed a spherical shape, with sizes ranging from 50 to 200 nm. A method was developed for conjugating OMVs obtained from broth and agar cultures of the typical strain and genovariants of V. cholerae O1 El Tor with FITC-labeled monoclonal antibodies (mAbs) against the outer membrane protein OmpU. The optimal OMVs concentrations providing a clear fluorescent signal were determined. The interaction of V. cholerae outer membrane vesicles with target cells was established using fluorescent mAbs and fluorescently labeled vesicles. OMVs were shown to aggregate and bind to the cell plasma membrane within 20 minutes. Internalization began within an hour. After three hours, virtually all vesicles penetrated the target cells. It was noted that OMVs produced by the typical strain and genovariants of V. cholerae O1 El Tor isolated from broth and agar cultures were internalized by target cells and accumulated intracellularly within identical timeframes. Under the experimental conditions used, no LPS involvement was detected in the internalization of V. cholerae O1 El Tor vesicles by target cells. Using cell culture models of two intestinal epithelial cell lines, Caсo-2 and HuTu 80, OMVs were shown to enhance the ability of bacterial cells to adhere to intestinal epithelial cells.References
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