E. A. Borobova*, D. V. Antonets, E. V. Starostina, L. I. Karpenko, A. A. Ilyichev and S. I. Bazhan Pages 375 - 385 ( 11 )
Objective: Immunotherapy based on induction of T-cell responses is a promising approach to cancer treatment. The study aims to design artificial epitope-based immunogens, DNA vaccine candidates against melanoma and evaluate their ability to stimulate tumor cytotoxicity of ex vivo generated T-cells.
Methods: The original computational methods were used for predicting T-cell epitopes and designing polyepitope melanoma antigens. Artificial genes encoding the target antigens were cloned into DNA vaccine plasmid vector. Target gene expression was confirmed both at transcriptional and translational level in HEK-293T cells transfected with DNA-vaccine constructs. Dendritic cells were generated from adherent peripheral blood mononuclear cells of HLA-A*02:01+ donors. Cytotoxic activity of effector lymphocytes stimulated in co-culture with autologous antigen-presenting dendritic cells towards melanoma Mel Is cells was assessed with lactate dehydrogenase release assay. The proportion of granzyme B producing CD8+ T-cells was estimated using intracellular cytokine staining and flow cytometry.
Results: Two DNA vaccine constructions were created - pMEL-TCI and pMEL-A0201 - encoding polypeptides containing T-cell epitopes of six immunodominant melanoma antigens (NY-ESO-1, MART1, MAGE-A1, MAGE-A11, MAGE-A3, and MAGE-C1). Dendritic cells transfected with DNA vaccine constructs were found to stimulate both tumor cytotoxicity mediated by autologous lymphocytes and granzyme B production by CD8+ T-cells, and pMEL-A0201 was found to be the most efficient.
Conclusion: The described approach may become a common platform for designing immunotherapeutic vaccines against oncological diseases.
Melanoma, Anti-tumor DNA vaccine, T-cell epitopes, Artificial polyepitope antigens, Cytotoxic response, MAGE.
State Research Center of Virology and Biotechnology , State Research Center of Virology and Biotechnology , State Research Center of Virology and Biotechnology , State Research Center of Virology and Biotechnology , State Research Center of Virology and Biotechnology , State Research Center of Virology and Biotechnology