Received: August 31, 2004, Accepted: November 18, 2004, Published online: July 1, 2005.

IMMUNIZATION WITH NATIVE AND COBALT 60-IRRADIATED Crotalus durissus terrificus VENOM IN SWISS MICE: ASSESSMENT OF THE NEUTRALIZING POTENCY OF ANTISERA

FERREIRA JUNIOR R. S. (1,2), NASCIMENTO N. (3), MARTINEZ J. C. (4),

ALVES J. B. (3), MEIRA D. A. (1), BARRAVIERA B. (1,2).

(1) Department of Tropical Diseases, Botucatu School of Medicine, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil; (2) Center for the Study of Venoms and Venomous Animals - CEVAP, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil; (3) Radiobiology Supervision - Nuclear Energy Research Institute (IPEN/CNEN/SP), São Paulo, Brazil; (4) Department of Production Engineering, School of Engineering, São Paulo State University, UNESP, Bauru, São Paulo, Brazil.

ABSTRACT: ELISA was used to evaluate, follow, and compare the humoral immune response of Swiss mice during hyperimmunization with natural and Cobalt 60-irradiated (60Co) Crotalus durissus terrificus venom. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by “in vivo” challenge and the side effects were assessed. The animals immunization with one LD50 of the venom was on days one, 15, 21, 30, and 45, when blood samples were collected; the challenges occurred on the 60th day. Results showed that ELISA was efficient in evaluating, following, and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60Co irradiation. Serum produced from Crotalus durissus terrificus irradiated venom showed higher potency and neutralization capacity than that from natural venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom.

KEY WORDS: hyperimmunization, irradiation, Crotalus durissus terrificus, ELISA, serum neutralization, detoxification.

CORRESPONDENCE TO:

R. S. FERREIRA JÚNIOR, Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, UNESP, Caixa Postal 577, 18618-000, Botucatu, São Paulo, Brasil. Email: rseabra@cevap.org.br