Role of crotoxin, a phospholipase A₂ isolated from Crotalus durissus terrificus snake venom, on inflammatory and immune reactions

Abstract

Background: Crotoxin (CTX) is a potent neurotoxin from Crotalus durissus terrificus snake venom (CdtV) composed of two subunits: one without catalytic activity (crotapotin), and a basic phospolipase A₂. Recent data have demonstrated that CdtV or CTX inhibit some immune and inflammatory reactions.Aim: The aim of this paper was to investigate the mechanisms involved in these impaired responses.Materials and methods: Male Swiss mice were bled before and at different intervals of time after subcutaneous injection of CTX or bovine serum albumin (BSA) (control animals). The effect of treatments on circulating leukocyte mobilisation and on serum levels of interleukin (IL)-6, IL-10, interferon (IFN)-γ and corticosterone were investigated. Spleen cells from treated animals were also stimulated in vitro with concanavalin A to evaluate the profile of IL-4, IL6, IL-10 or IFN-γ secretion. Cytokine levels were determined by immunoenzymatic assay and corticosterone levels by radioimmunoassay. To investigate the participation of endogenous corticosteroid on the effects evoked by CTX, animals were treated with metyrapone, an inhibitor of glucocorticoid synthesis, previous to CTX treatment.Results: Marked alterations on peripheral leukocyte distribution, characterised by a drop in the number of lymphocytes and monocytes and an increase in the number of neutrophils, were observed after CTX injection. No such alteration was observed in BSAtreated animals. Increased levels of IL-6, IL-10 and corticosterone were also detected in CTX-injected animals. IFN- γ levels were not modified after treatments. In contrast, spleen cells obtained from CTX-treated animals and stimulated with concanavalin A secreted less IL-10 and IL-4 in comparison with cells obtained from control animals. Metyrapone pretreatment was effective only to reverse the neutrophilia observed after CTX administration.Conclusions: Our results suggest that CTX may contribute to the deficient inflammatory and immune responses induced by crude CdtV.CTX induces endogenous mechanisms that are responsible, at least in part, for these impaired responses.