20.500.12556/RUL-137238 Combined TLR-3/TLR-8 signaling in the presence of α-type-1 cytokines represents a novel and potent dendritic cell type-1, anti-cancer maturation protocol During the ex vivo generation of anti-cancer dendritic cell (DC)-based vaccines, their maturation still represents one of the most crucial steps of the manufacturing process. A superior DC vaccine should: possess extensive expression of co-stimulatory molecules, have an exceptional type-1 polarization capacity characterized by their ability to produce IL-12p70 upon contact with responding T cells, migrate efficiently toward chemokine receptor 7 (CCR7) ligands, and have a superior capacity to activate cytotoxic T cell responses. A major advance has been achieved with the discovery of the next generation maturation protocol involving TLR-3 agonist (poly I:C), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, interferon (IFN)-γ, and IFN-α, and has since been known as α-type-1 maturation cocktail. We demonstrate how this combination can be greatly enhanced by the inclusion of a TLR-8 stimulation (R848), thereby contributing to potentiation between different TLR signaling pathways. For maximum efficiency, TLR-3 stimulation should precede (termed pre I:C) the stimulation with the R848/TNF-α/IL-1β/IFN-α/IFN-γ cocktail. When compared to DCs matured with α-type-1 maturation cocktail (αDCs), DCs matured with pre I:C/R848/TNF-α/IL-1β/IFN-α/IFN-γ (termed zDCs) displayed higher expression of CD80 and CD86 co-stimulatory molecules. Importantly, after CD40-ligand stimulation, which simulates DC-T cell contact, zDCs were much more proficient in IL-12p70 production. In comparison to αDCs, zDCs also displayed a significantly greater migratory capacity toward chemokine ligands (CCL)19 and CCL21, and had a significantly greater allo-stimulatory capacity. Finally, zDCs were also superior in their capacity to induce melanoma-specific CD8+ T cells, CD8+ T cell proliferation, and cytotoxic T cells, which produced approximately two times more IFN-γ and more granzyme B, than those stimulated with αDCs. In conclusion, we present a novel and superior DC maturation cocktail that could be easily implemented into next generation DC vaccine manufacturing protocols in future trials. dendritic cells type-1 polarization cytotoxic T cells maturation dendritične celice polarizacija tipa 1 citotoksične T celice true false true Angleški jezik Slovenski jezik Članek v reviji 2022-06-08 09:13:30 2022-06-08 09:13:36 2022-09-15 04:14:50 0000-00-00 00:00:00 2022 0 0 16 str. iss. 5, art. 835 Vol. 11 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616-006 2073-4409 10.3390/cells11050835 99744771 cells-11-00835-v2.pdf cells-11-00835-v2.pdf 1 6972CA5851F0DC4A8DC6A09F630682C3 7559b7893f583aa1832dd264a04556e1aaa1f51cfcd244e547a5d7cd6d952db3 8feca767-e6fa-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=157124 https://www.mdpi.com/2073-4409/11/5/835 1 5afc3a0b-e6fa-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=157123 Fakulteta za farmacijo 0 0 0