THE EFFECT OF CLOSTRIDIUM-DIFFICILE TOXIN ON COLONOCYTE PROSTANOID ACTIVITY

Antibiotic-associated colitis is caused by Clostridium difficile toxin. However, the pathophysiology of this entity is poorly understood. The aim of this study was to determine the effects of C. difficile toxin on colonocyte cyclooxygenase and phospholipase A(2) (PLA(2)) activity. A transformed colonocyte cell line (Caco-2) was grown to confluency on 6 well plates. The cells were stimulated with graded concentrations of C. difficile toxin. In separate experiments, the cells were pretreated for one hour prior to stimulation with the cyclooxygenase inhibitor, indomethacin, or the glucocorticoid, dexamethasone. The culture media was collected one hour following C. difficile stimulation. Prostaglandin E(2) (PGE(2)), 6-keto prostaglandin F-1 alpha (6KPGF), thromboxane B-2 (TxB(2)) and leukotriene B-4 (LTB(4)) levels were determined in the media by an ELISA. Platelet activating factor (PAF) concentration was determined by a RIA. C. difficile toxin stimulated PGE(2) and 6KPGF levels in a dose dependent fashion but failed to stimulate TxB(2), LTB(4) or PAF. Prostanoid production was inhibited by indomethacin dose dependently but was not inhibited by dexamethasone. The presence of indomethacin resulted in production of PAF. Our results show that the effects of C. difficile toxin on colonocytes are mediated by cyclooxygenase activity. The increase in PAF formation associated with indomethacin administration suggests that the prostanoids modulate PLA(2) activity and inhibit PAF formation.