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>Journals >Evidencia Médica e Investigación en Salud >Year 2015, Issue 3

Castañón-Sánchez CA, Enríquez-Rincón F, Figueroa-Arredondo P
Cytotoxic effect of the Vibrio cholerae cytolysin (VCC) on HEK293 cells
Evid Med Invest Salud 2015; 8 (3)

Language: Español
References: 20
Page: 111-116
PDF: 359.34 Kb.

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The non-O1 Vibrio cholerae group is composed by many diverse strains that are non-cholera toxin producers (CT -) or non-toxigenic strains), but still they are diarrheagenic and cytotoxic. These characteristics are attributed to the presence of a pore-forming toxin: the V. cholerae cytolysin (also known as VCC), since it is the best candidate to cause diarrhea in the human host. Characterization of the VCC as a vacuolation-inducer toxin was previously conducted by our group in tumorigenic and immortalized cell lines. In order to continue the study of the cytotoxicity of the VCC, we have chosen the HEK-293 cell line as a model of study that would help to further determine the role of the VCC as a virulence factor in the non O1 V. cholerae pathogenesis, since these cells are used for foreign protein expression from plasmids with the cloned genes. In vitro challenges were performed applying sterile bacterial supernatants rich in VCC (1:20) to HEK-293 cells, and later on, a microscopic search for cytotoxicity was performed. Here we show that there is a cytotoxic effect on HEK-293 cells, directly related to VCC, the toxin hyper-produced by V. cholerae non-O1 strains from our strain collection.

Key words: Vibrio cholerae, cytolysin, pore-forming toxin, HEK-293 cells.


  1. World Health Organization. Cholera. 2012. Available in: http://www.who.int/mediacentre/facthseets/fs107/en/

  2. Zitzer A, Wassenaar TM, Walev I, Bhakdi S. Potent membrane-permeabilizing cytocidal action of Vibrio cholerae cytolysin on human intestinal cells. Infect Immun. 1997; 65: 1293-1298.

  3. Figueroa-Arredondo P, Heuser JE, Akopyants NS, Morisaki JH, Giono-Cerezo S, Enríquez-Rincón F et al. Cell vacuolation caused by Vibrio cholerae hemolysin. Infect Immun. 2001; 69: 1613-1624.

  4. Olivier V, Queen J, Satchell KJ. Successful small intestine colonization of adult mice by Vibrio cholerae requires ketamine anesthesia and accessory toxins. PLoS One. 2009; 4 (10): e7352.

  5. De S, Olson R. Crystal structure of the Vibrio cholerae cytolysin heptamer reveals common features among disparate pore-forming toxins. Proc Natl Acad Sci. 2011; 108: 7385-7390.

  6. Yamamoto K, Ichinose Y, Shinagawa H, Makino K, Nakata A, Iwanaga M et al. Two step processing for activation of the cytolysin/hemolysin of Vibrio cholerae O1 biotype El Tor: nucleotide sequence of the structural gene (hlyA) and characterization of the processed products. Infect Immun. 1990; 58: 4106-4116.

  7. Dutta S, Mazumdar B, Banerjee KK, Ghosh AN. Three-dimensional structure of different functional forms of the Vibrio cholerae hemolysin oligomer: a cryo-electron microscopic study. J Bacteriol. 2010; 192: 169-178.

  8. Olson R, Gouaux E. Crystal structure of the Vibrio cholerae cytolysin (VCC) pro-toxin and its assembly into a heptameric transmembrane pore. J Mol Biol. 2005; 350: 997-1016.

  9. Paul K, Chattopadhyay K. Single point mutation in Vibrio cholerae cytolysin compromises the membrane pore-formation mechanism of the toxin. FEBS J. 2012; 279: 4039-4051.

  10. Zitzer AO, Nakisbekov NO, Li AV, Semiotrochev VL, Kiseliov YL, Muratkhodjaev JN et al. Entero-cytolysin (EC) from Vibrio cholerae non-O1 (some properties and pore-forming activity). Zentralbl Bakteriol. 1993; 279: 494-504.

  11. De Bernard M, Burroni D, Papini E, Rappuoli R, Telford J, Montecucco C. Identification of the Helicobacter pylori VacA toxin domain active in the cell cytosol. Infect Immun. 1998; 66 (12): 6014-6016.

  12. Mitra R, Figueroa P, Mukhopadhyay AK, Shimada T, Takeda Y, Berg DE et al. Cell vacuolation, a manifestation of the El Tor hemolysin of Vibrio cholerae. Infect Immun. 2000; 68: 1928-1933.

  13. Saka HA, Bidinost C, Sola C, Carranza P, Collino C, Ortiz S et al. Vibrio cholerae cytolysin is essential for high enterotoxicity and apoptosis induction produced by a cholerae toxin gene-negative V. cholera non-O1, non-O139 strain. Microb Phatog. 2008; 44: 118-128.

  14. Moschioni M, Tombola F, de Bernard M, Coelho A, Zitzer A, Zoratti M et al. The Vibrio cholerae haemolysin anion channel is required for cell vacuolation and death. Cel Microbiol. 2002; 4: 397-409.

  15. Rader AE, Murphy JR. Nucleotide sequences and comparation of the hemolysin determinants of Vibrio cholerae El Tor RV79(Hly+) and RV79(Hly-) and classical 569B(Hly-). Infect Immun. 1988; 56: 1414-1419.

  16. Honda T, Finkelstein RA. Purification and characterization of a hemolysin produced by Vibrio cholerae biotype El Tor: another toxic substance produced by cholera vibrios. Infect Immun. 1979; 26: 1020-1027.

  17. Coelho A, Andrade JR, Vicente ACP, DiRita VJ. Cytotoxic cell vacuolation activity from Vibrio cholerae hemolysin. Infect Immun. 2000; 68: 1700-1705.

  18. Saka HA, Gutiérrez MG, Bocco JL, Colombo MI. The autophagic pathway: a cell survival strategy against the bacterial pore-forming toxin Vibrio cholerae cytolysin. Autophagy. 2007; 3 (4): 363-365.

  19. Levan S, De S, Olson R. Vibrio cholerae cytolysin recognizes the heptasaccharide core of complex N-glycans with nanomolar affinity. J Mol Biol, 2013; 424: 944-957.

  20. Vidal EJ, Enríquez-Rincón F, Giono-Cerezo S, Ribas-Aparicio RM, Figueroa-Arredondo P. Culture supernants from V. cholerae O1 El Tor strains isolated from different geographic areas induce cell vacuolation and cytotoxicity. Salud Pública Mex. 2009; 51: 39-47.

>Journals >Evidencia Médica e Investigación en Salud >Year 2015, Issue 3

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