Метод получения рекомбинантной идиотипической ДНК-вакцины против лимфом

Abstract

Idiotype, the unique part of immunoglobulin molecule expressed on the surface of B-cells, represents a specific antigen for vaccination against lymphoma. Method of PCR-based amplification of variable immunoglobulin regions was developed and optimized. Three immunoglobulin expressing lymphoma cell lines IM-9, Daudi, Namalwa and tumor specimens from two pediatric patients with B-cell non-Hodgkin lymphoma were used for idiotype cloning. We performed design of specific primers for direct amplification of all possible variable regions of IgM, IgG heavy chains, IgK, IgL light chains and described method of cloning, assembling scFv and expression of idiotypic protein in bacteria E. coli. We add a co-stimulator gene of potato virus X coat protein (PVXCP) to the construction in the N-part of the protein. The proposed method of vaccine production is suitable for the production of vaccines for immune therapy in more than 96% of cases of B-cell non-Hodgkin lymphoma.