Longus, a Type IV Pilus of Enterotoxigenic Escherichia Coli, is Involved in Adherence to Intestinal Epithelial Cells

Enterotoxigenic Escherichia coli (ETEC) is the leading bacterial cause of diarrhea in the developing world, as well as the most common cause of traveler's diarrhea. The main hallmarks of this type of bacteria are the expression of one or more enterotoxins and fimbriae used for attachment to host intestinal cells. Longus is a pilus produced by ETEC. These bacteria grown in pleuropneumonia-like organism (PPLO) broth at 37 degrees C and in 5% CO(2) produced longus, showing that the assembly and expression of the pili depend on growth conditions and composition of the medium. To explore the role of longus in the adherence to epithelial cells, quantitative and qualitative analyses were done, and similar levels of adherence were observed, with values of 111.44 x 10(4) CFU/ml in HT-29, 101.33 x 10(4) CFU/ml in Caco-2, and 107.11 x 10(4) CFU/ml in T84 cells. In addition, the E9034A Delta lngA strain showed a significant reduction in longus adherence of 32% in HT-29, 22.28% in Caco-2, and 21.68% in T84 cells compared to the wild-type strain. In experiments performed with nonintestinal cells (HeLa and HEp-2 cells), significant differences were not observed in adherence between E9034A and derivative strains. Interestingly, the E9034A and E9034A Delta lngA(pLngA) strains were 30 to 35% more adherent in intestinal cells than in nonintestinal cells. Twitching motility experiments were performed, showing that ETEC strains E9034A and E9034A Delta lngA(pLngA) had the capacity to form spreading zones while ETEC E9034A Delta lngA does not. In addition, our data suggest that longus from ETEC participates in the colonization of human colonic cells.