PRODUCTION OF BETULA PENDULA POLLEN Bet v 2 ALLERGEN WITH IMMUNOREGULATORY SIGNALS AND ITS RECOGNITION BY IgE ANTIBODIES

Background. Creation of recombinant proteins, containing profilin and signal peptides which provide proteasomal and lysosomal degradation pathways of proteins. Materials and methods. Plasmid pET-Bet v 2, encoding Bet v 2 allergen, was used as a base vector. Two signal peptides: C-terminal fragment of ornithine decarboxylase (ODCsig) and N-terminal fragment of human invariant chain (Ii) — were cloned into this vector. Recombinant proteins were produced in BL21 (DE3) strain of E. coli. The recombinant protein was purified using NiNTA agarose. Immunological specificity of the recombinant proteins was estimated by means of immune-enzyme methods using sera of patients suffering from birch pollen allergy. Results. Bet v 2 allergen (profilin) of Betula pendula pollen was modified by immunoregulatory peptides in order to change the IgE/IgG response ratio. Ornithine decarboxylase proteasome degradation signal was used to enhance the Th1 response and human invariant chain signal directed to lysosomes was used in order to enhance Th2 response. The production system of recombinant allergen Bet v 2 preparation suitable for immunological tests was developed. The produced highly purified chimeric proteins preparations bound IgE antibodies of allergic to birch pollen patients' sera efficiently. Conclusion. The new chimeric allergens based on Bet v 2 protein of Betula pendula pollen were created and isolated.

Betula pendula, Bet v 2, profilm, immunoregulatory signals