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Infection and Immunity, January 2001, p. 52-57, Vol. 69, No. 1
Department of Molecular Biosciences,
University of Kansas, Lawrence, Kansas 66045-2106
Received 14 July 2000/Accepted 20 September 2000
Enterotoxigenic Escherichia coli (ETEC) is capable of
invading epithelial cell lines derived from the human ileum and colon. Two separate invasion loci (tia and tib) that
direct noninvasive E. coli strains to adhere to and invade
cultured human intestine epithelial cells have previously been isolated
from the classical ETEC strain H10407. The tib locus
directs the synthesis of TibA, a 104-kDa outer membrane glycoprotein.
Synthesis of TibA is directly correlated with the adherence and
invasion phenotypes of the tib locus, suggesting that this
protein is an adhesin and invasin. Here we report the purification of
TibA and characterization of its biological activity. TibA was purified
by continuous-elution preparative sodium dodecyl sulfate-polyacrylamide
gel electrophoresis. Purified TibA was biotin labeled and then shown to
bind to HCT8 human ileocecal epithelial cells in a specific and
saturable manner. Unlabeled TibA competed with biotin-labeled TibA,
suggesting the presence of a specific TibA receptor in HCT8 cells.
These results show that TibA acts as an adhesin. Polyclonal anti-TibA
antiserum inhibited invasion of ETEC strain H10407 and of recombinant E. coli bearing tib locus clones, suggesting
that TibA also acts as an invasin. The ability of TibA to direct
epithelial cell adhesion suggests a role for this protein in ETEC pathogenesis.
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.1.52-57.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Enterotoxigenic Escherichia coli TibA
Glycoprotein Adheres to Human Intestine Epithelial Cells
and
*
Corresponding author. Mailing address: University of
Kansas, Department of Molecular Biosciences, 7049 Haworth Hall,
Lawrence, KS 66045-2106. Phone: (785) 864-4299. Fax: (785)
864-5294. E-mail: elsingh{at}ukans.edu.
Present address: Institut fuer Tropenmedizin Universitaet
Tuebingen, D-72074 Tuebingen, Germany.
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