title: The role of toxin A and toxin B in Clostridium difficile infection.
abstract: Clostridium difficile infection is the leading cause of healthcare-associated diarrhoea in Europe and North America. During infection, C. difficile produces two key virulence determinants, toxin A and toxin B. Experiments with purified toxins have indicated that toxin A alone is able to evoke the symptoms of C. difficile infection, but toxin B is unable to do so unless it is mixed with toxin A or there is prior damage to the gut mucosa. However, a recent study indicated that toxin B is essential for C. difficile virulence and that a strain producing toxin A alone was avirulent. This creates a paradox over the individual importance of toxin A and toxin B. Here we show that isogenic mutants of C. difficile producing either toxin A or toxin B alone can cause fulminant disease in the hamster model of infection. By using a gene knockout system to inactivate the toxin genes permanently, we found that C. difficile producing either one or both toxins showed cytotoxic activity in vitro that translated directly into virulence in vivo. Furthermore, by constructing the first ever double-mutant strain of C. difficile, in which both toxin genes were inactivated, we were able to completely attenuate virulence. Our findings re-establish the importance of both toxin A and toxin B and highlight the need to continue to consider both toxins in the development of diagnostic tests and effective countermeasures against C. difficile.
authors.affiliation: Clostridia Research Group, Centre for Biomolecular Sciences, School of Molecular Medical Sciences, Nottingham Digestive Diseases Centre, NIHR Biomedical Research Unit, University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Kuehne
authors.forename: Stephen T
authors.initials: ST
authors.lastname: Cartman
authors.forename: John T
authors.initials: JT
authors.lastname: Heap
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Kelly
authors.forename: Alan
authors.initials: A
authors.lastname: Cockayne
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Minton
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D016360:Clostridioides difficile / Q000145:classification / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D003015:Clostridium Infections / Q000382:microbiology*; D006224:Cricetinae; D004195:Disease Models, Animal; D004768:Enterotoxins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D019073:HT29 Cells; D006801:Humans; D009500:Neutralization Tests; D014709:Vero Cells; D014774:Virulence / Q000235:genetics
chemical_list: D057134:Antibodies, Neutralizing; D001427:Bacterial Toxins; C488583:Clostridium difficile lethal toxin B; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile