Snake venom has been the subject of numerous studies in an attempt to find properties and biological effects that may be beneficial to man. In this study we evaluated in vitro the effects of Crotalus durissus terrificus (Cdt) and Crotalus durissus collilineatus (Cdc) venom in human peripheral blood mononuclear cells (PBMCs). At 24？h, a significant decrease of viable cells was observed in cells stimulated with the Cdc venom at 0.0005？mg/mL and 0.005？mg/mL compared to the negative control. At 48？h, a significant decrease of viable cells was observed only in cells stimulated with Cdc venom at 0.005？mg/mL. A significant increase of TNF-α and IL-10 was detected 48 hours after culture of PBMC with Cdc, but not with Cdt venom. The expression of CD69 and PD1 (programmed death-1), activation and regulatory cell markers, on CD8+ and CD8？ T cells did not change in the presence of Cdt and Cdc venom. Our results suggest the presence of proinflammatory and anti-inflammatory components in the Cdc venom. Further analysis should be done to identify those Cdc venom components as it has been done for the Cdt venom by other authors, indicating that modulatory components are found in the venom of different species of Crotalus snakes. 1. Introduction Snakes are included in one of the most diverse groups of reptiles in the world, with over 3100 known living species [1]. Brazil has cataloged 721 species of snakes. Only two families (Elapidae and Viperidae) show venomous species, those that produce toxins in specialized glands [2]. Snakes belonging to Viperidae family and Crotalus genus are represented in Brazil by a single species, Crotalus durissus and five subspecies known as Crotalus durissus terrificus (Cdt), Crotalus durissus collilineatus (Cdc), Crotalus durissus cascavella, Crotalus durissus ruruima, and Crotalus durissus marajoensis. The genus Crotalus, which is popularly known as rattlesnakes, whose venom is the subject of this study, has been highlighted in the research and its venom has the highest lethality rate [3–5]. Snake venom is a complex mixture consisting of proteins, peptides, amino acids, nucleotides, lipids, and carbohydrates that have a wide variety of biological activities that reflect changes in the biological properties and toxicity [6, 7]. The venom of the rattlesnakes is considered one of the most toxic venom from Brazilian snakes. The high toxicity is due to the presence of various toxins [8], mainly convulxin, crotamine, crotoxin, and gyroxin [9]. Despite all the toxic effects, the venom usually is a rich source of molecules with important
References
[1]
P. Uetz and J. Hallerman, “The TIGR Reptile Database,” 2009, http://www.reptile-database.org.
[2]
Sociedade Brasileira de Herpetologia—SBH, “Lista Brasileira de Anfíbios e Répteis. Disponível em,” 2011, http://www.sbherpetologia.org.br/.
[3]
G. Rosenfeld, “Acidentes por animais pe？onhentos,” in Doen？as Infecciosas e Parasitárias, R. Veronesi, Ed., pp. 951–962, Rio de Janeiro, Guanabara Koogan, 8th edition, 1991.
[4]
I. M. B. Francischetti, M. E. C. Gombarovits, J. G. Valenzuela, C. R. Carlini, and J. A. Guimar？es, “Intraspecific variation in the venoms of the South American rattlesnake (Crotalus durissus terrificus),” Comparative Biochemistry and Physiology C, vol. 127, no. 1, pp. 23–36, 2000.
[5]
J. L. C. Cardoso, F. O. S. Fran？a, F. Wen, S. Málaque, and H. V. Junior, Animais Pe？onhentos no Brasil, Biologia, Clínica e Terapêutica dos Acidentes, Sarvier, S？o Paulo, Brazil, 2th edition, 2009.
[6]
A. H. Cruz, R. Z. Mendon？a, and V. L. Petricevich, “Crotalus durissus terrificus venom interferes with morphological, functional, and biochemical changes in murine macrophage,” Mediators of Inflammation, vol. 2005, no. 6, pp. 349–359, 2005.
[7]
D. A. Warrell, “Snake bite,” The Lancet, vol. 375, no. 9708, pp. 77–88, 2010.
[8]
Y. Oshima-Franco, S. Hyslop, J. Prado-Franceschi, and M. A. Cruz-H？fling, “Neutralizing capacity of antisera raised in horses and rabbits against Crotalus durissus terrificus (South American rattlesnake) venom and its main toxin, crotoxin,” Toxicon, vol. 37, no. 10, pp. 1341–1357, 1999.
[9]
A. Rangel-Santos, E. C. Dos-Santos, M. Lopes-Ferreira, C. Lima, D. F. Cardoso, and I. Mota, “A comparative study of biological activities of crotoxin and CB fraction of venoms from Crotalus durissus terrificus, Crotalus durissus cascavella and Crotalus durissus collilineatus,” Toxicon, vol. 43, no. 7, pp. 801–810, 2004.
[10]
K. Stocker, “Use of snake venom proteins in medicine,” Schweizerische Medizinische Wochenschrift, vol. 129, no. 6, pp. 205–216, 1999.
[11]
D. C. Koh, A. Armugam, and K. Jeyaseelan, “Snake venom components and their applications in biomedicine,” Cellular and Molecular Life Sciences, vol. 63, no. 24, pp. 3030–3041, 2006.
[12]
W. Rajendra, A. Armugam, and K. Jeyaseelan, “Neuroprotection and peptide toxins,” Brain Research Reviews, vol. 45, no. 2, pp. 125–141, 2004.
[13]
I. F. Purchase, P. A. Botham, L. H. Bruner, O. P. Flint, J. M. Frazier, and W. S. Stokes, “Workshop overview: scientific and regulatory challenges for the reduction, refinement, and replacement of animals in toxicity testing,” Toxicological Sciences, vol. 43, no. 2, pp. 86–101, 1998.
[14]
A. Hernández Cruz, S. Garcia-Jimenez, R. Zucatelli Mendon？a, and V. L. Petricevich, “Pro- and anti-inflammatory cytokines release in mice injected with Crotalus durissus terrificus venom,” Mediators of Inflammation, vol. 2008, Article ID 874962, 10 pages, 2008.
[15]
D. F. Cardoso, M. Lopes-Ferreira, E. L. Faquim-Mauro, M. S. Macedo, and S. H. P. Farsky, “Role of crotoxin, a phospholipase A2 isolated from Crotalus durissus terrificus snake venom, on inflammatory and immune reactions,” Mediators of Inflammation, vol. 10, no. 3, pp. 125–133, 2001.
[16]
B. C. Favoretto, R. Ricardi, S. R. Silva et al., “Immunomodulatory effects of crotoxin isolated from Crotalus durissus terrificus venom in mice immunised with human serum albumin,” Toxicon, vol. 57, no. 4, pp. 600–607, 2011.
[17]
A. Rangel-Santos, C. Lima, M. Lopes-Ferreira, and D. F. Cardoso, “Immunosuppresive role of principal toxin (crotoxin) of Crotalus durissus terrificus venom,” Toxicon, vol. 44, no. 6, pp. 609–616, 2004.
[18]
D. Aderka, “The potential biological and clinical significance of the soluble tumor necrosis factor receptors,” Cytokine and Growth Factor Reviews, vol. 7, no. 3, pp. 231–240, 1996.
[19]
L. Thompson-Snipes, V. Dhar, M. W. Bond, T. R. Mosmann, K. W. Moore, and D. M. Rennick, “Interleukin 10: a novel stimulatory factor for mast cells and their progenitors,” Journal of Experimental Medicine, vol. 173, no. 2, pp. 507–510, 1991.
[20]
S. F. Ziegler, F. Ramsdell, and M. R. Alderson, “The activation antigen CD69,” Stem Cells, vol. 12, no. 5, pp. 456–465, 1994.
[21]
E. H. Neto, A. L. J. Coelho, A. L. F. Sampaio et al., “Activation of human T lymphocytes via integrin signaling induced by RGD-disintegrins,” Biochimica et Biophysica Acta, vol. 1773, no. 2, pp. 176–184, 2007.
[22]
A. H. Sharpe, E. J. Wherry, R. Ahmed, and G. J. Freeman, “The function of programmed cell death 1 and its ligands in regulating autoimmunity and infection,” Nature Immunology, vol. 8, no. 3, pp. 239–245, 2007.
[23]
E. A. Said, F. P. Dupuy, L. Trautmann et al., “Programmed death-1-induced interleukin-10 production by monocytes impairs CD4+ T cell activation during HIV infection,” Nature Medicine, vol. 16, no. 4, pp. 452–459, 2010.
[24]
K. W. Moore, A. O’Garra, R. Dewaalmalefyt, P. Vieira, and T. R. Mosmann, “Interleukin-10,” Annual Review of Immunology, vol. 11, pp. 165–190, 1993.
[25]
K. Asadullah, W. Sterry, and H. D. Volk, “Interleukin-10 therapy—review of a new approach,” Pharmacological Reviews, vol. 55, no. 2, pp. 241–269, 2003.
