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LamB‐mediated adherence of enteropathogenic Escherichia coli to HEp‐2 cells
Author(s) -
Subramanian K.,
Shankar R.B.,
Meenakshisundaram S.,
Lakshmi B.S.,
Williams P.H.,
Balakrishnan A.
Publication year - 2008
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2008.03800.x
Subject(s) - enteropathogenic escherichia coli , microbiology and biotechnology , escherichia coli , enterobacteriaceae , biology , genetics , gene
Aims: To establish the role of maltoporin (LamB) in adherence of enteropathogenic Escherichia coli (EPEC) to epithelial cells in vitro. Methods and Results: Three strains, wild type (WT) EPEC, a maltoporin (LamB) mutant ΔlamB , and DH5α were used to study adherence to cultured HEp‐2 cells. Mutant ΔlamB was found to be deficient in adherence compared to WT EPEC. Adherence of ΔlamB was restored to wild type levels when complemented with the cloned lamB gene. The non–adherent strain DH5α also adhered to HEp‐2 cells when it harboured the cloned lamB gene. The LamB protein was isolated from WT EPEC by electroelution and antibodies were raised in rabbits. The specificity of the antibodies was analysed by Western blotting. Anti‐LamB antiserum reduced adherence of WT EPEC to HEp‐2 cells. The LamB protein was coated on latex beads and the beads adhered to HEp‐2 cells. Anti‐LamB antiserum prevented bead adherence to HEp‐2 cells. Multiple sequence alignment showed that the L9 loop of EPEC LamB had four amino acids different from the L9 loop of LamB from several other related pathogens. Conclusions: LamB serves as an alternative or additional adherence factor for EPEC. Significance and Impact of the Study: Adherence is an important component of the pathogenesis of noninvasive pathogens like EPEC. A putative adhesin such as LamB, which has already been found to be co‐expressed with virulence factor EspB may be a potential vaccine candidate for control of EPEC and related pathogens.