Inflammation induced by Bothrops asper venom: release of proinflammatory cytokines and eicosanoids, and role of adhesion molecules in leukocyte infiltration Stella Regina Zamuner a , Juliana Pavan Zuliani a , Cristina Maria Fernandes a , Jose ´ Maria Gutie ´rrez b , Catarina de Fa ´tima Pereira Teixeira a, * a Laborato ´rio de Farmacologia, Instituto Butantan, Ave Vital Brasil 1500, Sa ˜o Paulo, SP, CEP 05503-900, Brazil b Facultad de Microbiologı ´a, Instituto Clodomiro Picado, Universidad de Costa Rica, San Jose ´, Costa Rica Received 16 May 2005; revised 17 August 2005; accepted 18 August 2005 Available online 29 September 2005 Abstract Bothrops asper venom (BaV) causes systemic and local effects characterized by an acute inflammatory reaction with accumulation of leukocytes and release of endogenous mediators. In this study, the effects of BaV on the release of the cytokines IL-1, IL-6 and TNF-a and the eicosanoids LTB 4 and TXA 2 in the peritoneal cavity of mice were analyzed. We also investigated the participation of b 2 integrin chain, L-selectin, LFA-1, ICAM-1 and PECAM-1 adhesion molecules in the BaV-induced leukocyte accumulation. Levels of proinflammatory cytokines IL-6 and TNF-a, as well as eicosanoids LTB 4 and TXA 2 were significantly increased after BaV injection (250 mg/kg), whereas no increment in IL-1 was observed. Anti-mouse L-selectin, LFA-1, ICAM-1, PECAM-1 and b 2 integrin chain monoclonal antibodies resulted in a reduction of neutrophil accumulation induced by BaV injection compared with isotype-matched control injected animals. These data suggest that BaV is able to induce the activation of leukocytes and endothelium to express adhesion molecules involved in the recruitment of neutrophils into the inflammed site. Furthermore, these results showed that BaV induces the release of cytokines and eicosanoids in the local of the venom injection; these inflammatory mediators may be important for the initiation and amplification of the inflammatory reaction characteristic from Bothrops sp envenomation. q 2005 Elsevier Ltd. All rights reserved. Keywords: Snake venom; Adhesion molecules; Cytokines; Eicosanoids; Inflammation 1. Introduction In Latin America, the majority of snakebites are caused by species of the genus Bothrops, and local effects are the most frequent consequence of these envenomations, which are poorly neutralized by conventional antivenom ser- otherapy (Avila-Aguero et al., 2001). In severe cases, these local effects may lead to permanent tissue loss, disability or amputation (Cardoso et al., 1993; Theakston and Theakston, 1996). The pathogenesis of venom-induced local effects is rather complex, involving the combined action of venom components, such as proteinases and phospholipases (Gutierrez and Lomonte, 1995; Bjarnason and Fox, 1994). Bothrops asper is the most important poisonous snake in Central America (Gutierrez and Lomonte, 1995). Besides inducing hemorrhage and myonecrosis, several studies demonstrate that B. asper venom evokes a complex inflammatory reaction characterized by multimediated Toxicon 46 (2005) 806–813 www.elsevier.com/locate/toxicon 0041-0101/$ - see front matter q 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.toxicon.2005.08.011 * Corresponding author. Fax: C55 11 3726 1505. E-mail address: cteixeir@usp.br (C.F. Pereira Teixeira).