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Infection and Immunity, August 1999, p. 4019-4026, Vol. 67, No. 8
0019-9567/99/$04.00+0

Characterization of an Enterotoxigenic Escherichia coli Strain from Africa Expressing a Putative Colonization Factor

Sami B. Khalil,1 Frederick J. Cassels,2 Hind I. Shaheen,1 Lewis K. Pannell,3 Nemat El-Ghorab,1 Karim Kamal,1 Moustafa Mansour,1 Stephen J. Savarino,1 and Leonard F. Peruski Jr.1,*

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- 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.


Infection and Immunity, August 1999, p. 4019-4026, Vol. 67, No. 8
0019-9567/99/$04.00+0



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  • Anantha, R. P., McVeigh, A. L., Lee, L. H., Agnew, M. K., Cassels, F. J., Scott, D. A., Whittam, T. S., Savarino, S. J. (2004). Evolutionary and Functional Relationships of Colonization Factor Antigen I and Other Class 5 Adhesive Fimbriae of Enterotoxigenic Escherichia coli. Infect. Immun. 72: 7190-7201 [Abstract] [Full Text]