ACTIVATION OF CELLULAR PHOSPHOLIPASE-A2 BY CLOSTRIDIUM-DIFFICILE TOXIN-B

C difficile toxin B is a potent cytotoxin known to disrupt the microfi laments of cultured cells. We have recently shown also increased phosp holipase A2 activity in cells treated with toxin B. The activity was d etected as a toxin-induced, dose-dependent release of C-14-arachidonic acid from prelabeled fibroblasts. Here is shown that the toxin elicit ed a C-14-arachidonic acid release in a cell mutant resistant to the t oxin B effect on the microfilaments. The toxin-induced release was fur ther characterized using fibroblasts. Within 20 min high doses of toxi n B (6 mug/ml) elicited a release which increased exponentially with t ime. Of the major membrane phospholipids the lipase activity affected mainly phosphatidyl ethanolamine. Neither cycloheximide nor pertussis toxin treatment of target cells inhibited the toxin-induced release, w hile it could be increased with 12-O-tetradecanoylphorbol-13-acetate. Our results also suggest a toxin-mediated increase in phospholipase C activity occurring at a later stage than the phospholipase A2 activati on. We conclude that the ability of toxin B to induce phospholipase ac tivation represents a hitherto unrecognized toxin B effect which is ne ither a cause nor a consequence of toxin-induced microfilament disorga nization.