title: Clostridium difficile toxin B is more potent than toxin A in damaging human colonic epithelium in vitro.
abstract: Toxin A but not toxin B, appears to mediate intestinal damage in animal models of Clostridium difficile enteritis. The purpose of this study was to investigate the electrophysiologic and morphologic effects of purified C. difficile toxins A and B on human colonic mucosa in Ussing chambers. Luminal exposure of tissues to 16-65 nM of toxin A and 0.2-29 nM of toxin B for 5 h caused dose-dependent epithelial damage. Potential difference, short-circuit current and resistance decreased by 76, 58, and 46%, respectively, with 32 nM of toxin A and by 76, 55, and 47%, respectively, with 3 nM of toxin B, when compared with baseline (P < 0.05). 3 nM of toxin A did not cause electrophysiologic changes. Permeability to [3H]mannitol increased 16-fold after exposure to 32 nM of toxin A and to 3 nM of toxin B when compared with controls (P < 0.05). Light and scanning electron microscopy after exposure to either toxin revealed patchy damage and exfoliation of superficial epithelial cells, while crypt epithelium remained intact. Fluorescent microscopy of phalloidin-stained sections showed that both toxins caused disruption and condensation of cellular F-actin. Our results demonstrate that the human colon is approximately 10 times more sensitive to the damaging effects of toxin B than toxin A, suggesting that toxin B may be more important than toxin A in the pathogenesis of C. difficile colitis in man.
authors.affiliation: University Clinic of Surgery, Vienna, Austria.
authors.forename: M
authors.initials: M
authors.lastname: Riegler
authors.forename: R
authors.initials: R
authors.lastname: Sedivy
authors.forename: C
authors.initials: C
authors.lastname: Pothoulakis
authors.forename: G
authors.initials: G
authors.lastname: Hamilton
authors.forename: J
authors.initials: J
authors.lastname: Zacherl
authors.forename: G
authors.initials: G
authors.lastname: Bischof
authors.forename: E
authors.initials: E
authors.lastname: Cosentini
authors.forename: W
authors.initials: W
authors.lastname: Feil
authors.forename: R
authors.initials: R
authors.lastname: Schiessel
authors.forename: J T
authors.initials: JT
authors.lastname: LaMont
mesh_terms: D000199:Actins / Q000187:drug effects / Q000378:metabolism; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000633:toxicity*; D001692:Biological Transport / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D016360:Clostridioides difficile; D003106:Colon; D003603:Cytotoxins / Q000633:toxicity; D004594:Electrophysiology; D004768:Enterotoxins / Q000633:toxicity*; D004848:Epithelium / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D007700:Kinetics; D008353:Mannitol / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D008855:Microscopy, Electron, Scanning; D013997:Time Factors
chemical_list: D000199:Actins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D003603:Cytotoxins; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile; D008353:Mannitol