Leukocyte response induced by Bothrops jararaca crude venom: in vivo and in vitro studies

The effect of Bothrops jararaca crude venom (BjV) on the cellular component of inflammatory responses was investigated in vivo and in vitro. In vivo leukocyte accumulation and release of eicosanoids (thromboxane A2, TXA2, and leukotriene B4, LTB4) at the site of injection of the venom were assessed using the air pouch method in rats. Administration of BjV caused a significant cell accumulation, maximal values being obtained after 6-8 hr. Neutrophils were the predominant cell type in the inflammatory exudate. High concentrations of LTB4 were detected 1-4 hr after the injection of the venom. TXA2 concentrations were significantly increased only at the early stages of the response to the venom. In vitro chemotaxis assays were performed and showed that the venom per se was not able to induce oriented neutrophil migration because varying concentrations of the venom dissolved in Hanks' balanced salt solution (HBSS) evoked a response equivalent to that of HBSS alone. Furthermore, the venom did not affect cellular intrinsic mechanisms involved with neutrophil locomotion because previous incubation of the cells with BjV produced no effect. However, high concentrations of the venom were able to generate serum chemotactic factor(s). Incubation of serum with the venom evoked a neutrophil migration similar to that observed with serum activated by lipopolysaccharide from Escherichia coli. Participation of chemotactic factors derived from the complement system is suggested by data showing loss of this activity when serum was heated (56 degrees C) before the addition of BjV. The present results suggest that leukocyte accumulation in the locality of a lesion induced by BjV is dependent on secretion or activation of endogenous components responsible for several steps in leukocyte recruitment instead of a direct effect of the venom on leukocytes.