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To activate tumor antigen specific CD4+ helper and CD8+ cytotoxic T lymphocyte responses genetically modified dendritic cells (DC) are increasingly used. In this study, we used electro-transfection of DCs with hTERT mRNA that enables an HLA independent whole antigen approach potentially targeting a wide range of hTERT epitopes. Immature, i.e. non-proliferating human DCs were prepared from peripheral blood monocytes in serum-free growth medium, GM-CSF and IL-4. Subsequently the DCs were electroporated in transfection buffer with mRNA and matured in a cytokine cocktail consisting of IL-1beta, IL-6, TNF-alpha and PGE2.

Efficient in vitro expression of human reverse transcriptase (hTERT) in dendritic cells of lung cancer patients using RNA electrotransfection