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Infection and Immunity, August 1999, p. 4019-4026, Vol. 67, No. 8
Research Sciences Department, U.S. Naval
Medical Research Unit No. 3, Cairo, Egypt1;
Department of Enteric Infections, Division of Communicable
Diseases and Immunology, Walter Reed Army Institute of Research,
Washington, D.C. 20307-51002; and
Structural Mass Spectrometry Group, National Institute of
Diabetes and Digestive and Kidney Diseases, National Institutes of
Health, Bethesda, Maryland 208923
Received 19 January 1999/Returned for modification 3 March
1999/Accepted 20 May 1999
An enterotoxigenic Escherichia coli (ETEC) strain of
serotype O114:H
0019-9567/99/$04.00+0
Characterization of an Enterotoxigenic
Escherichia coli Strain from Africa Expressing a Putative
Colonization Factor
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 Mr 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°C but not when grown at 22°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.
*
Corresponding author. Mailing address: c/o Commanding
Officer, U.S. Naval Medical Research Unit No. 3, PSC 452, Box 5000, FPO
AE 09835 0007. Phone: 011-20/2-284-1381. Fax: 011-20/2-284-7121. E-mail: boushrah{at}namru3.navy.mil.
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