Abstract
Adhesion is the first step in the pathogenesis of enterotoxigenic Escherichia coli infections. The genes encoding the most prevalent adhesion factors CFA/I, CS3 and CS6 were cloned into Vibrio cholerae strain CVD 103–HgR and expression of fimbriae was investigated in wildtype and recombinant strains by transmission electron microscopy in conjunction with immunolabelling and negative staining. Negative staining was effective in revealing CFA/I and CS3, but not CS6. Although morphology of fimbriae differed between wildtype and recombinant strains, corresponding surface antigens were recognized by specific antibodies. The present study provides evidence that ETEC-specific fimbriae can adequately be expressed in an attenuated V. cholerae vaccine strain and that immunoelectron microscopy is a critical tool to validate the surface expression of antigens in view of their possible suitability for recombinant vaccines.
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Burnette WN (1981) “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203
de Lorimier AJ, Byrd W, Hall ER et al (2003) Murine antibody response to intranasally administered enterotoxigenic Escherichia coli colonization factor CS6. Vaccine 21:2548–2555
Ehara M, Ishibashi M, Ichinose Y et al (1987) Purification and partial characterization of fimbriae of Vibrio cholerae O1. Vaccine 5:283–288
Evans DG, Evans DJ Jr, Clegg S et al (1979) Purification and characterization of the CFA/I antigen of enterotoxigenic Escherichia coli. Infect Immun 25:738–748
Favre D, Lüdi S, Stoffel M et al (2006) Expression of enterotoxigenic Escherichia coli colonization factors in Vibrio cholerae. Vaccine 24:4354–4368
Hall RH, Vial PA, Kaper JB et al (1988) Morphological studies on fimbriae expressed by Vibrio cholerae 01. Microb Pathog 4:257–265
Jiang ZD, Lowe B, Verenkar MP et al (2002) Prevalence of enteric pathogens among international travelers with diarrhea acquired in Kenya (Mombasa), India (Goa), or Jamaica (Montego Bay). J Infect Dis 185:497–502
Jonson AB, Normark S, Rhen M (2005) Fimbriae, pili, flagella and bacterial virulence. Contrib Microbiol 12:67–89
Ketley JM, Michalski J, Galen J et al (1993) Construction of genetically marked Vibrio cholerae O1 vaccine strains. FEMS Microbiol Lett 111:15–21
Knutton S, Lloyd DR, Candy DC et al (1985) Adhesion of enterotoxigenic Escherichia coli to human small intestinal enterocytes. Infect Immun 48:824–831
Knutton S, Lloyd DR, McNeish AS (1987) Identification of a new fimbrial structure in enterotoxigenic Escherichia coli (ETEC) serotype O148:H28 which adheres to human intestinal mucosa: a potentially new human ETEC colonization factor. Infect Immun 55:86–92
Knutton S, McConnell MM, Rowe B et al (1989) Adhesion and ultrastructural properties of human enterotoxigenic Escherichia coli producing colonization factor antigens III and IV. Infect Immun 57:3364–3371
Levine MM, Ristaino P, Marley G et al (1984) Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans. Infect Immun 44:409–420
Nishikawa Y, Helander A, Ogasawara J et al (1998) Epidemiology and properties of heat-stable enterotoxin-producing Escherichia coli serotype O169:H41. Epidemiol Infect 121:31–42
Novotny C, Carnahan J, Brinton CC Jr (1969) Mechanical removal of F pili, type I pili, and flagella from Hfr and RTF donor cells and the kinetics of their reappearance. J Bacteriol 98:1294–1306
Oyofo BA, Subekti DS, Svennerholm AM et al (2001) Toxins and colonization factor antigens of enterotoxigenic Escherichia coli among residents of Jakarta, Indonesia. Am J Trop Med Hyg 65:120–124
Sohanpal BK, Kulasekara HD, Bonnen A et al (2001) Orientational control of fimE expression in Escherichia coli. Mol Microbiol 42:483–494
Acknowledgments
The technical support of Cynthia Furer, Yvonne Schlatter, Corinne Hug and Simon König is gratefully acknowledged. We also acknowledge M. Wolf, Walter Reed Army Hospital, Maryland, USA for generously providing pM295 and pM392 and Fred Cassels, NIH, Bethesda, Maryland, USA for providing purified antigens. Supported by the Federal Office for Professional Education and Technology OPET (Grant KTI 7304.1 LSPP-LS).
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Lüdi, S., Frey, J., Favre, D. et al. Morphological and immunocytochemical analysis of Escherichia coli-specific surface antigens in wildtype strains and in recombinant Vibrio cholerae . Antonie van Leeuwenhoek 93, 185–192 (2008). https://doi.org/10.1007/s10482-007-9192-0
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DOI: https://doi.org/10.1007/s10482-007-9192-0