Cp. Kelly et al., NEUTROPHIL RECRUITMENT IN CLOSTRIDIUM-DIFFICILE TOXIN-A ENTERITIS IN THE RABBIT, The Journal of clinical investigation, 93(3), 1994, pp. 1257-1265
Neutrophil infiltration is a prominent feature of Clostridium difficil
e-associated enteritis and colitis. The aim of this study was to exami
ne the importance of neutrophil recruitment and neutrophil-mediated ti
ssue damage in C. difficile toxin A-induced enteritis. Competitive bin
ding experiments using purified H-3-toxin A demonstrated the presence
of a single class of medium affinity receptors on rabbit neutrophils (
K-d 7 X 10(-8) M). Pertussis toxin and the nonhydrolyzable GTP analog
GTPgamma S both inhibited H-3-toxin A binding (by 56 and 65%, respecti
vely), indicating that the rabbit neutrophil toxin A receptor is G pro
tein linked. Toxin A elicited a dose-dependent (25-200 mu g/ml) stimul
ation of neutrophil migration in vitro, and this functional effect was
also pertussis toxin sensitive (69% inhibition). Treatment of neutrop
hils with R15.7, a blocking monoclonal antibody to the leukocyte adhes
ion molecule CD18, inhibited toxin A-stimulated neutrophil migration b
y 85% in vitro. Pretreatment of rabbits with R15.7 also prevented neut
rophil infiltration of toxin A-exposed ileal loops in vivo as determin
ed by histologic examination and by ileal tissue myeloperoxidase level
s. Furthermore, R15.7 effected a substantial inhibition of fluid secre
tion (by 65%), mannitol permeability (by 66%), and histologic damage i
n toxin A-exposed ileal loops. Anti-CD18 (R15.7) had no inhibitory eff
ect on cholera toxin enterotoxicity. These data demonstrate that C. di
fficile toxin A is a proinflammatory toxin whose enterotoxic effects a
re substantially dependent upon neutrophil recruitment.