Dendritic cell (DC) immunotherapy is capable of generating tumour-specific immune responses. Different maturation strategies were previously tested to obtain DC capable of anti-cancer responses in vitro, usually with limited clinical benefit. Mutual comparison of currently used maturation strategies and subsequent complex evaluation of DC functions and their stimulatory capacity on T cells was performed in this study to optimize the DC vaccination strategy for further clinical application. DC were generated from monocytes using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4, pulsed with whole tumour cell lysate and then matured with one of five selected maturation strategies or cultured without additional maturation stimulus. DC were characterized with regard to their surface marker expression, cytokine profiles, migratory capacity, allogeneic and autologous T cell stimulatory capacity as well as their specific cytotoxicity against tumour antigens. We were able to demonstrate extensive variability among different maturation strategies currently used in DC immunotherapeutic protocols that may at least partially explain limited clinical benefit of some clinical trials with such DC. We identified DC matured with interferon-γ and lipopolysaccharide as the most attractive candidate for future clinical trials in cancer immunotherapy.

• MeSH
• biologické markery metabolismus   MeSH
• cytokiny metabolismus   MeSH
• dendritické buňky imunologie   MeSH
• faktor stimulující granulocyto-makrofágové kolonie metabolismus   farmakologie   MeSH
• imunoterapie metody   MeSH
• interleukin-10 metabolismus   MeSH
• interleukin-12 metabolismus   MeSH
• interleukin-4 metabolismus   farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• monocyty cytologie   MeSH
• nádorové buněčné linie MeSH
• nádory imunologie   terapie   MeSH
• pohyb buněk MeSH
• T-lymfocyty imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• cytokiny MeSH
• faktor stimulující granulocyto-makrofágové kolonie MeSH
• IL10 protein, human MeSH Prohlížeč
• interleukin-10 MeSH
• interleukin-12 MeSH
• interleukin-4 MeSH

BACKGROUND: For clinical applications, dendritic cells (DCs) need to be generated using GMP-approved reagents. In this study, we tested the characteristics of DCs generated in two clinical grade culture media and activated by three maturation stimuli, Poly I: C, LPS and the mixture of proinflammatory cytokines in order to identify the optimal combination of culture media and activation stimulus for the clinical use. METHOD: We tested DCs generation using two GMP-certified culture media, CellGro and RPMI+5% human AB serum and evaluated DCs morphology, viability and capapability to mature. We tested three maturation stimuli, PolyI:C, LPS and the mixture of proinflammatory cytokines consisting of IL-1, IL-6, TNF and prostaglandin E2. We evaluated the capacity of activated DCs to induce antigen-specific T cells and regulatory T lymphocytes. RESULTS: Cell culture in CellGro resulted in a higher yield of immature DCs resulting from increased number of adherent monocytes. DCs that were generated in CellGro and activated using Poly I:C were the most efficient in expanding antigen-specific T cells compared to the DCs that were generated in other media and activated using LPS or the cocktail of proinflammatory cytokines. A comparison of all tested combinations revealed that DCs that were generated in CellGro and activated using Poly I:C induced low numbers of regulatory T cells. CONCLUSION: In this study, we identified monocyte-derived DCs that were generated in CellGro and activated using Poly I:C as the most potent clinical-grade DCs for the induction of antigen-specific T cells.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• dendritické buňky cytologie   účinky léků   MeSH
• epitopy imunologie   MeSH
• fenotyp MeSH
• imunoterapie metody   MeSH
• klinické zkoušky jako téma MeSH
• kultivační média farmakologie   MeSH
• lidé MeSH
• nádory imunologie   terapie   MeSH
• poly I-C farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• regulační T-lymfocyty cytologie   účinky léků   imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• epitopy MeSH
• kultivační média MeSH
• poly I-C MeSH