Characterization of an enterotoxigenic Escherichia coli strain from Africa expressing a putative colonization factor

Infect Immun. 1999 Aug;67(8):4019-26. doi: 10.1128/IAI.67.8.4019-4026.1999.

Abstract

An enterotoxigenic Escherichia coli (ETEC) strain of serotype O114:H- that expressed both heat-labile and heat-stable enterotoxins and tested negative for colonization factors (CF) was isolated from a child with diarrhea in Egypt. This strain, WS0115A, induced hemagglutination of bovine erythrocytes and adhered to the enterocyte-like cell line Caco-2, suggesting that it may elaborate novel fimbriae. Surface-expressed antigen purified by differential ammonium sulfate precipitation and column chromatography yielded a single protein band with M(r) 14,800 when resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (16% polyacrylamide). A monoclonal antibody against this putative fimbrial antigen was generated and reacted with strain WS0115A and also with CS1-, CS17-, and CS19-positive strains in a dot blot assay. Reactivity was temperature dependent, with cells displaying reactivity when grown at 37 degrees C but not when grown at 22 degrees C. Immunoblot analysis of a fimbrial preparation from strain WS0115A showed that the monoclonal antibody reacted with a single protein band. Electron microscopy and immunoelectron microscopy revealed fimbria-like structures on the surface of strain WS0115A. These structures were rigid and measured 6.8 to 7.4 nm in diameter. Electrospray mass-spectrometric analysis showed that the mass of the purified fimbria was 14,965 Da. The N-terminal sequence of the fimbria established that it was a member of the CFA/I family, with sequence identity to the amino terminus of CS19, a new CF recently identified in India. Cumulatively, our results suggest that this fimbria is CS19. Screening of a collection of ETEC strains isolated from children with diarrhea in Egypt found that 4.2% of strains originally reported as CF negative were positive for this CF, suggesting that it is biologically relevant in the pathogenesis of ETEC.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigens, Bacterial / isolation & purification*
  • Bacterial Adhesion
  • Bacterial Proteins / isolation & purification*
  • Cattle
  • Diarrhea / etiology
  • Escherichia coli / immunology*
  • Escherichia coli / pathogenicity
  • Female
  • Fimbriae Proteins*
  • Fimbriae, Bacterial / ultrastructure
  • Humans
  • Infant
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Electron
  • Molecular Sequence Data
  • Molecular Weight

Substances

  • Antigens, Bacterial
  • Bacterial Proteins
  • colonization factor antigens
  • Fimbriae Proteins