Title:Design of Artificial Immunogens Containing Melanoma-associated T-cell Epitopes
VOLUME: 18 ISSUE: 6
Author(s):E.A. Borobova*, D.V. Antonets, E.V. Starostina, L.I. Karpenko, A.A. Ilyichev and S.I. Bazhan
Affiliation:State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk Region, State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk Region, State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk Region, State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk Region, State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk Region, State Research Center of Virology and Biotechnology "Vector", Koltsovo, Novosibirsk Region
Keywords:Melanoma, anti-tumor DNA vaccine, T-cell epitopes, artificial polyepitope antigens, cytotoxic response, MAGE.
Abstract: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.
