Multiple elements controlling adherence of enterohemorrhagic Escherichia coli O157: H7 to HeLa cells

Alfredo Torres, James B. Kaper

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

Adherence of enterohemorrhagic Escherichia coli (EHEC) to the intestinal epithelium is essential for initiation of infection. Intimin is the only factor demonstrated to play a role in intestinal colonization by EHEC O157:H7. Other attempts to identify additional adhesion factors in vitro have been unsuccessful, suggesting that expression of these factors is under tight regulation. We sought to identify genes involved in the control of adherence of EHEC O157:H7 to cultured epithelial cells. A total of 5,000 independent transposon insertion mutants were screened for their ability to adhere to HeLa cells, and 7 mutants were isolated with a markedly enhanced adherence. The mutants adhered at levels 113 to 170% that of the wild-type strain, and analysis of the protein profiles of these mutants revealed several proteins differentially expressed under in vitro culture conditions. We determined the sequence of the differentially expressed proteins and further investigated the function of OmpA, whose expression was increased in a mutant with an insertionally inactivated tcdA gene. An isogenic ompA mutant showed reduced adherence compared to the parent strain. Disruption of the ompA gene in the tdcA mutant strain abolished the hyperadherent phenotype, and anti-OmpA serum inhibited adhesion of wild-type and tdcA mutant strains to HeLa cells. Enhanced adhesion mediated by OmpA was also observed with Caco-2 cells, and anti-OmpA serum blocked adherence to HeLa cells of other EHEC O157:H7 strains. Our results indicate that multiple elements control adherence and OmpA acts as an adhesin in EHEC O157:H7.

Original languageEnglish (US)
Pages (from-to)4985-4995
Number of pages11
JournalInfection and Immunity
Volume71
Issue number9
DOIs
StatePublished - Sep 1 2003

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Enterohemorrhagic Escherichia coli
Escherichia coli O157
HeLa Cells
Genes
Caco-2 Cells
Mutant Proteins
Intestinal Mucosa
Serum
Cultured Cells
Proteins
Epithelial Cells
Phenotype
Infection

ASJC Scopus subject areas

  • Immunology

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Multiple elements controlling adherence of enterohemorrhagic Escherichia coli O157 : H7 to HeLa cells. / Torres, Alfredo; Kaper, James B.

In: Infection and Immunity, Vol. 71, No. 9, 01.09.2003, p. 4985-4995.

Research output: Contribution to journalArticle

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