加热对尖吻蝮蛇蛇毒及其组分免疫原性及生物毒性影响-effect of heat on immunogenicity and biological toxicity of agkistrodon acutus venom and its components.docxVIP

Effects of heating on the immunogenicity and biological toxicity of Deinagkistrodon acutus venom and its fractionsABSTRACTBackground:Serum therapy is the primary treatment for systemic envenomation by D. acutus. Three types of D. acutus antivenom are produced and available in China: liquid antivenom from Shanghai Institute of Biological Products, lyophilized antivenom from Cheng-Du Military Area Center of Disease Prevention and Control, and lyophilized antivenom from the National Institute of Preventive Medicine, Taipei, Taiwan. Antivenom is isolated from the plasma of adult horses immunized with formaldehyde-detoxified D. acutus venom. Although formaldehyde or glutaraldehyde-treated venom can produce high-titerantiserum[1,2,3], repeated immunization of the formaldehyde- or glutaraldehyde-containingsolution inevitably harms the horse. Venom detoxification methods are needed that produce antivenom with higher immunogenicity and lower toxicity. Several improved techniques for venom detoxification have been reported, including titration of native venom with iodine monochloride solution[4], encapsulation of native crotoxin in liposomes[5], irradiation of toxic proteins with gamma-rays[6,7], and selective heat denaturation[8,9]. Heat denaturation is the simplest detoxification technique and it has the ability to reduce venomtoxicity without altering the immunogenicity of components with molecular weights lower than 25 kDa[8,9].Objectives:The aim of this study was to assess the effects of various temperatures on the precipitation, immunogenicity, biological toxicity, and interactions of different toxic components from D. acutus venom, and improve toxoid preparation.Methods:Separation of fractionsThe D. acutus venom was separated by gel filtration chromatography (Sephadex G-50), eluted by distilled water with a flow rate of 2.0 ml/min at room temperature (25oC).The target fractions were pooled and lyophilized. Then their LD50 and hemorrhage acticity were determined.Deter