title: Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro.
abstract: BACKGROUND
  Dendritic cells (DC) have been used for immunotherapy of malignant tumors, different kinds of infections, and other clinical conditions. For that purpose, optimal conditions for the generation of functionally mature DC in vitro are required. Two different protocols for the induction of maturation of monocyte-derived DC (MDDC) were compared in this study.
  
  
  METHODS
  MDDC were generated in vitro by cultivating adherent monocytes of healthy volunteers with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) during 6-days period. The immature DC thus prepared were induced to mature using two protocols. DC were stimulated for 2 days with lipopolysaccharide (LPS), or with a cocktail of proinflammatory mediators (PM) containing IL-1beta, IL-6, tumor necrosis factor alpha (TNFalpha), and prostaglandin E2 (PGE2), respectively. Phenotypic characteristics of MDDC and their endocytic activity were studied by flow cytometry. Allostimulatory activity of these cells was tested in the mixed leukocyte reaction (MLR), whereas the production of cytokines was determined by ELISA kits.
  
  
  RESULTS
  MDDC matured with PM (PM-DC) were predominantly non-adherent cells, while about 30% of LPS-matured DC were adherent cells. In comparison with LPS-DC, PM-DC expressed higher levels of CD86 and CD83, had lower endocytic activity, produced higher levels of IL-10 and lower levels of IL-12, and more strongly stimulated proliferation of allogeneic lymphocytes.
  
  
  CONCLUSIONS
  The protocol based on the combination of proinflammatory cytokines and PGE2 is better for the induction of maturation of human MDDC in vitro than the protocol using LPS alone.
authors.affiliation: Military Medical Academy, Institute for Medical Research, Belgrade, Serbia & Montenegro. vmaimi@eunet.yu
authors.forename: Miodrag
authors.initials: M
authors.lastname: Colić
authors.forename: Slavko
authors.initials: S
authors.lastname: Mojsilović
authors.forename: Bojan
authors.initials: B
authors.lastname: Pavlović
authors.forename: Dragana
authors.initials: D
authors.lastname: Vucićević
authors.forename: Ivana
authors.initials: I
authors.lastname: Majstorović
authors.forename: Biljana
authors.initials: B
authors.lastname: Bufan
authors.forename: Zorica
authors.initials: Z
authors.lastname: Stojić-Vukanić
authors.forename: Sasa
authors.initials: S
authors.lastname: Vasilijić
authors.forename: Dragana
authors.initials: D
authors.lastname: Vucević
authors.forename: Sonja
authors.initials: S
authors.lastname: Gasić
authors.forename: Bela
authors.initials: B
authors.lastname: Balint
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000502:physiology; D015232:Dinoprostone / Q000494:pharmacology; D004705:Endocytosis; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D018836:Inflammation Mediators / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008213:Lymphocyte Activation; D009000:Monocytes / Q000166:cytology; D010641:Phenotype
chemical_list: D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D015232:Dinoprostone