title: Both enzymatic and non-enzymatic properties of heat-labile enterotoxin are responsible for LT-enhanced adherence of enterotoxigenic Escherichia coli to porcine IPEC-J2 cells.
abstract: Previous studies in piglets indicate that heat labile enterotoxin (LT) expression enhances intestinal colonization by K88 adhesin-producing enterotoxigenic Escherichia coli (ETEC) as wild-type ETEC adhered to intestinal epithelium in substantially greater numbers than did non-toxigenic constructs. Enzymatic activity of the toxin was also shown to contribute to the adhesion of ETEC and non-ETEC bacteria to epithelial cells in culture. To further characterize the contribution of LT to host cell adhesion, a nontoxigenic, K88-producing E. coli was transformed with either the gene encoding for LT holotoxin, a catalytically-attenuated form of the toxin [LT(R192G)], or LTB subunits, and resultant changes in bacterial adherence to IPEC-J2 porcine intestinal epithelial cells were measured. Strains expressing LT holotoxin or mutants were able to adhere in significantly higher numbers to IPEC-J2 cells than was an isogenic, toxin-negative construct. LT+ strains were also able to significantly block binding of a wild-type LT+ ETEC strain to IPEC-J2 cells. Adherence of isogenic strains to IPEC-J2 cells was unaltered by cycloheximide treatment, suggesting that LT enhances ETEC adherence to IPEC-J2 cells independent of host cell protein synthesis. However, pretreating IPEC-J2 cells with LT promoted adherence of negatively charged latex beads (a surrogate for bacteria which carry a negative change), which adherence was inhibited by cycloheximide, suggesting LT may induce a change in epithelial cell membrane potential. Overall, these data suggest that LT may enhance ETEC adherence by promoting an association between LTB and epithelial cells, and by altering the surface charge of the host plasma membrane to promote non-specific adherence.
authors.affiliation: Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD 57007, USA.
authors.forename: Peter Z
authors.initials: PZ
authors.lastname: Fekete
authors.forename: Kristina S
authors.initials: KS
authors.lastname: Mateo
authors.forename: Weiping
authors.initials: W
authors.lastname: Zhang
authors.forename: Rodney A
authors.initials: RA
authors.lastname: Moxley
authors.forename: Radhey S
authors.initials: RS
authors.lastname: Kaushik
authors.forename: David H
authors.initials: DH
authors.lastname: Francis
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000276:immunology; D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology*; D002460:Cell Line; D054307:Enterotoxigenic Escherichia coli / Q000201:enzymology / Q000378:metabolism*; D004768:Enterotoxins / Q000378:metabolism*; D004847:Epithelial Cells / Q000382:microbiology*; D004927:Escherichia coli Infections / Q000382:microbiology; D006358:Hot Temperature; D007413:Intestinal Mucosa / Q000382:microbiology; D013552:Swine
chemical_list: D018829:Adhesins, Bacterial; D004768:Enterotoxins