title: Day 3 Poly (I:C)-activated dendritic cells generated in CellGro for use in cancer immunotherapy trials are fully comparable to standard Day 5 DCs.
abstract: BACKGROUND
  Dendritic cells (DCs) are professional antigen-presenting cells that are capable of inducing immune responses. DC-based vaccines are normally generated using a standard 5- to 7-day protocol. To shorten the DC-based vaccine production for use in cancer immunotherapy, we have developed a fast DC protocol by comparing standard DCs (Day 5 DCs) and fast DCs (Day 3 DCs).
  
  
  METHODS
  We tested the generation of Day 5 versus Day 3 DCs using CellGro media and subsequent activation by two activation stimuli: Poly (I:C) and LPS. We evaluated DC morphology, viability, phagocyte activity, cytokine production and ability to stimulate antigen-specific T cells.
  
  
  RESULTS
  Day 5 and Day 3 DCs exhibited similar phagocytic capacity. Poly (I:C)-activated Day 5 DCs expressed higher levels of the costimulatory and surface molecules CD80, CD86 and HLA-DR compared to Poly (I:C)-activated Day 3 DCs. Nevertheless, LPS-activated Day 5 and Day 3 DCs were phenotypically similar. Cytokine production was generally stronger when LPS was used as the maturation stimulus, and there were no significant differences between Day 5 and Day 3 DCs. Importantly, Day 5 and Day 3 DCs were able to generate comparable numbers of antigen-specific CD8(+) T cells. The number of Tregs induced by Day 5 and Day 3 DCs was also comparable.
  
  
  CONCLUSIONS
  We identified monocyte-derived DCs generated in CellGro for 3 days and activated using Poly (I:C) similarly potent in most functional aspects as DCs produced by the standard 5 day protocol. These results provide the rationale for the evaluation of faster protocols for DC generation in clinical trials.
authors.affiliation: Department of Immunology, Charles University, Second Faculty of Medicine and University Hospital Motol, Prague, Czech Republic; Sotio, Prague, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Truxova
authors.affiliation: Sotio, Prague, Czech Republic.
authors.forename: Katerina
authors.initials: K
authors.lastname: Pokorna
authors.affiliation: Sotio, Prague, Czech Republic.
authors.forename: Kamila
authors.initials: K
authors.lastname: Kloudova
authors.affiliation: Department of Immunology, Charles University, Second Faculty of Medicine and University Hospital Motol, Prague, Czech Republic; Sotio, Prague, Czech Republic.
authors.forename: Simona
authors.initials: S
authors.lastname: Partlova
authors.affiliation: Department of Immunology, Charles University, Second Faculty of Medicine and University Hospital Motol, Prague, Czech Republic; Sotio, Prague, Czech Republic.
authors.forename: Radek
authors.initials: R
authors.lastname: Spisek
authors.affiliation: Department of Immunology, Charles University, Second Faculty of Medicine and University Hospital Motol, Prague, Czech Republic; Sotio, Prague, Czech Republic. Electronic address: jitka.fucikova@fnmotol.cz.
authors.forename: Jitka
authors.initials: J
authors.lastname: Fucikova
mesh_terms: D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D019496:Cancer Vaccines / Q000276:immunology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002470:Cell Survival; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy; D009369:Neoplasms / Q000276:immunology* / Q000378:metabolism / Q000628:therapy; D010587:Phagocytosis / Q000276:immunology; D010641:Phenotype; D011070:Poly I-C / Q000276:immunology* / Q000494:pharmacology; D037182:T-Cell Antigen Receptor Specificity / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D013997:Time Factors
chemical_list: D019496:Cancer Vaccines; D016207:Cytokines; D011070:Poly I-C
keywords: Cancer immunotherapy; Dendritic cell; Fast generation; Monocyte; Vaccine