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Ammodytoxin content of Vipera ammodytes ammodytes venom influences the level of venom-toxicity neutralising antibodies in venom immunised rabbits

Long-nosed viper (Vipera ammodytes ammodytes) venom (Vamam venom) has been used as an antigen for immunisation of animals (horses) in the process of antivenom (venom-specific antisera) production. Recently we have shown that the level of antibodies specific to Vamam venom's most toxic component - ammodytoxin A (AtxA) (anti-AtxA IgG) in Vamam venom immunised rabbit sera was highly correlated to their venom-toxicity neutralization potential (Halassy et al. Comp Biochem Physiol C 2008 ; 148, 178). Since all ammodytoxins are highly immuno-crossreactive, antibodies measured by anti-AtxA IgG ELISA might be considered as anti-Atx antibodies in total. Thus allows to hypothesize that the total Atx content of venom used for immunisation might be one of the critical factors determining the outcome of immunisation procedure. The aim of this study was to investigate the outcome of immunisation procedure for antivenom production in rabbits, when antigens for immunisation were venom batches with high or low ammodytoxin content, respectively. The venom batches used for immunisation were selected on the basis of previously noticed differences in Atx content by recently developed HPLC method (Halassy et al. J Liq Chromatogr RT 2008 ; 31, 38). The new sandwich ELISA was developed for precise determination of Atx content in Vamam venom and Atx quantified in venoms used for immunisation of rabbits. Results showed that indeed animals immunised with the venom containing lower amounts of Atx developed the significantly lower level of venom toxicity neutralising antibodies. The level of anti-AtxA IgG was again highly correlated to the venom-toxicity neutralising potential of immunised rabbit sera.

Vipera ammodytes ammodytes; Ammodytoxin; Haemorrhagin; Antivenom; ELISA; Toxicity-neutralization potency

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