engleski

Immune reaction specific for venom of long-nosed viper (V. ammodytes) in mice and influence of peptidoglycan type adjuvants

Long-nosed viper (V. ammodytes) is one of the most poisonous snakes and is widely distributed through central Europe. Consequences of its bite are serious problem in human as well in veterinarian medicine. Specific antivenin is the only available therapeutic drug for curing snake venom poisoning. Induction of specific immune reaction (IR) by gradual immunisation of big animals (horse, donkey, sheep) is the only available method for antivenin production. The aim of our work was to establish a mice model for studying IR specific to snake venom, that could be used for modifications in immunisation protocol to achieve the optimal response. These modifications could involve changes in quantities of venom used for immunisations, changes in immunisation schedule, use of adjuvants or liposome preparations. In preliminary experiments we found the maximal quantity of V. ammodytes venom that is not lethal for mice. Using that amount of venom (6,56mg/dose) we established the optimal immunisation schedule (four s.c. immunisations in three week intervals). The specific IR was monitored by ELISA for venom specific IgGs established in our laboratory. Gradual reduction of immunisation dose revealed that the same level of specific antibodies in serum can be obtained by half of the maximal dose. For studying the effect of adjuvants two structurally related compounds were used: peptidoglycan monomer GlcNAc-MurNAc-L-Ala-D-isoGln-mesoDAP(wNH2)-D-Ala-D-Ala (PGM) and L-(adamant-2-il)-Gly-L-Ala-D-isoGln (AdTP2). Suboptimal doses of venom (0,1 and 0,5mg) were used. Both adjuvants stimulated specific IR. In conclusion, mice model for studying the venom specific immune response has been established and adjuvant activity of PGM and AdTP2 in immune reaction to a very complex antigen, as the snake venom is, has been demonstrated.

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