Histopathology of peritoneal wall sections (serous membrane and skeletal muscle of the floor of the dorsal cavity) in mAb-treated animals (2 h) showed vasodilatation signs with expressive numbers of intravascular leukocytes (leukocytosis), edema, and discreet hemorrhage (Fig. 4A). Cavity samples from control animals were represented by accentuated endomisial edema with muscular fiber dissociation and moderate hemorrhage (Fig. 4B). In addition, some muscle fibers exhibited coagulation necrosis (hyalinized: without Osimertinib order striations and slightly eosinophilic). The pancreas from mice treated with mAbs exhibited hemorrhage and discreet edema in the intestine/pancreas interface (Fig. 4C). Conversely,

controls that received only B. atrox venom showed evidence of extensive solid hemorrhage and acinar cell dissociation in pancreatic samples using conventional microscopy ( Fig. 4D). Although Camargo et al. (2005) observed acute pancreatitis induced by phospholipase A2 from Bothrops venom in rats, the changes in the peritoneal cavity and pancreas found in our study are probably associated to the direct contact between the mAb and venom mixture injected into the peritoneal cavity. Kidney histopathology from animals treated with mAbs (2 h) was not significantly different from that of control

animals ( Fig. 4E, F). Although human deaths by Bothrops envenomation are generally associated to acute renal failure ( Milani Jr. et al., 1997), renal failure was not well reproduced in murine models. Moreover, several studies that evaluated renal this website alterations caused by bothropic venom in rats were performed

using i.v. PI3K inhibitor injection or ex-vivo renal perfusion ( Gutiérrez et al., 2009; Boer-Lima et al., 1999), and this could explain the lack of alterations in kidney samples evaluated in this study. Mice inoculated with the mAb and venom mixture lost the same quantity of blood as negative controls when bleeding time was determined (Fig. 5). In contrast, high blood loss was observed in mice given venom only. To our knowledge this is the first study to show that neutralizing monoclonal antibodies against three major Bothrops venom toxins abrogates the venom activity. Our results show that a pool of three mAbs neutralizes the lethal activity of B. atrox venom. Nevertheless, we believe that the action of toxins present in minor concentration in the venom ( Neiva et al., 2009), which could act alone or synergistically with other toxins, must also be considered. Moreover, intraspecific ( Núñez et al., 2009) and interspecific ( Queiroz et al., 2008) variation in venom characteristics should also be investigated when developing antivenoms based on monoclonal antibodies. Monoclonal antibodies similarly to polyclonal antibodies when injected into xenogeneic animals induce antibody production against either their constant and variable regions resulting in a short circulating life.