Adherence of enterotoxigenicKlebsiella pneumoniaeto hela cells and expression of type 3 fimbriae

Abstract

In 1986, an outbreak of diarrheal disease in Texas was attributed to contaminated iced tea in whichKlebsiella pneumoniaewas the predominant flora. Although the ten strains tested were of different serotypes, they all produced a heat labile enterotoxin. Some of these enterotoxigenic strains (ETKP) also exhibited mannose-resistant hemagglutinins, which were later found to be type 3 MR/K fimbriae. In this study we extended our observations and investigated fimbrial expression, as well as HeLa cell adherence behavior, using negative staining, immunogold labeling, and scanning electron microscopy (SEM).ETKP strains were grown on colonization factor antigen agar overnight at 37° C for expression of fimbriae. The presence of fimbriae and immunogold labeling of the cells was determined as described by Wolfeet al.Antiserum was prepared against purified fimbriae in rabbits and dilutions of primary and secondary antibodies (goat anti-rabbit IgG conjugated with 10 nm gold particles) in phosphate buffered saline, pH 7.2, containing 1 % bovine serum albumin and 0.5% Tween 20 were used to label cells.