Poly(I:C) used for human dendritic cell maturation preserves their ability to secondarily secrete bioactive IL-12.
Dendritic cells (DC) are professional antigen-presenting cells that play a central role in the control of immunity. Mature DC are characterized by high expression levels of MHC and co-stimulatory molecules, and by the secretion of IL-12, a key cytokine for the priming of cytotoxic T lymphocytes. Here, we have compared different maturation stimuli to reproducibly generate stable mature DC secreting high amounts of bioactive IL-12p70. We have compared soluble human trimeric CD40 ligand (sCD40L) combined with IFN-gamma, poly(I:C), a cocktail of cytokines (IL-1beta, IL-6 and tumor necrosis factor-alpha) with prostaglandin E(2) and lipopolysaccharide. A major concern, however, is whether DC, that have already produced high amounts of IL-12p70 during the maturation step, are still capable of secreting IL-12p70 after in vivo administration at the time of interaction with the targeted T cells. To mimic that situation, mature DC generated by those methods were compared for their ability to secrete IL-12p70 in the absence of IFN-gamma, using sCD40L. We observed a second consistent secretion of bioactive IL-12p70 upon subsequent sCD40L stimulation only when poly(I:C) was used as the maturating agent. Our data suggest that, for clinical use, poly(I:C) may be one of the most appropriate agents to generate stable mature DC. These mature DC might generate in vivo effective immune responses after injection, because they retain the ability to secrete bioactive IL-12 after CD40 ligation.
Rouas
Redouane
R
Laboratory of Experimental Hematology, Jules Bordet Institute, Free University of Brussels, 1000 Brussels, Belgium.
Lewalle
Philippe
P
El Ouriaghli
Frank
F
Nowak
Berengere
B
Duvillier
Hughes
H
Martiat
Philippe
P
D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D018664:Interleukin-12 / Q000096:biosynthesis*; D011070:Poly I-C / Q000276:immunology / Q000494:pharmacology*; D021122:Protein Subunits / Q000096:biosynthesis
D000954:Antigens, Surface; D021122:Protein Subunits; D018664:Interleukin-12; D011070:Poly I-C