Cytokine profile on the ehrlich ascites tumor treated with Bothrops jararaca venom

THESIS. R. J. da Silva submitted his thesis for his Doctor of Philosophy degree in Tropical Biology publicly examined at Botucatu School of Medicine of São Paulo State University, UNESP, Botucatu, in 2000.

Advisor. Profa. Dra. Denise Fecchio.

ABSTRACT. Bothrops jararaca venom shows a major antineoplastic effect on Ehrlich ascites tumor (EAT) growth. The objectives of this study were to evaluate the tumor growth kinetics, inflammatory influx, cytokine profile in the peritoneal cavity, and the in vitro antimitotic and/or toxic venom effect on tumor and peritoneal cells. Swiss adult male mice were injected with 1 x 10³ EAT cells in the peritoneal cavity and divided into two groups. One group received saline and the other 0.4 mg/kg of venom. Treatment consisted of five inoculations with 72-hour interval, starting 24 hours after tumor implantation. On the 2^nd, 5^th, 8^th, 11^th, and 14^th day of tumor growth, the animals were sacrificed to determine the number of tumor, polymorphonuclear (PMN), and mononuclear (MN) cells in the peritoneal cavity and IL-1a, IL-2, IL-4, IL-6, IL-10, IL-13, and TNF-a levels in the peritoneal washing supernatant by ELISA. Two other control groups, consisting of animals that were not injected with tumor cells were formed: one received saline and the other venom (0.4 mg/kg). These were analyzed using the same protocol as the EAT groups. In the in vitro study, EAT or peritoneal cells were cultivated in RPMI medium in 5% CO₂ at 37ºC with Bothrops jararaca venom at concentrations of 2.5, 5, 10, 20, 40, and 80 mg. Cell sample viability was determined 3, 6, 12, and 24 hours after cultivation. The results obtained in this study allow the following conclusions: 1) EAT implantation in mice induces high tumor growth rates, influx of PMN and MN leukocytes into the peritoneal cavity from the 8^th day of tumor growth, and TNF-a, IL-6, and IL-10 production on the 11^th and 14^th days; 2) Treatment of EAT mice with 0.4 mg/kg of Bothrops jararaca venom induces tumor growth inhibition, increased PMN and MN leukocyte influx into the peritoneal cavity at early stages of tumor growth (2^nd and 5^th days), and suppression of TNF-a, IL-6, and IL-10 production; and 3) The in vitro treatment with Bothrops jararaca venom produces a toxic effect on EAT and peritoneal cells, the higher toxicity being observed in the peritoneal cells.

KEY WORDS: Ehrlich ascites tumor, cytokines, Bothrops jararaca venom.

CORRESPONDENCE TO:

R. J. da SILVA - Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP/UNESP, Distrito de Rubião Júnior, S/N, CEP 18618-000, Botucatu, São Paulo, Brasil.

E-mail: [e-mail]

Publication Dates

• Publication in this collection
  02 Apr 2001
• Date of issue
  2001