title: Human T lymphocyte priming in vitro by haptenated autologous dendritic cells.
abstract: Dendritic cells (DC), generated from adherent peripheral blood mononuclear cells (PBMC) by culturing with granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4, were used to study in vitro sensitization of naive, hapten-specific T cells and to analyse cross-reactivities to related compounds. DC were hapten-derivatized with nickel sulphate (Ni) or 2-hydroxyethyl-methacrylate (HEMA), followed by tumour necrosis factor-alpha (TNF-alpha)-induced maturation, before autologous T cells and a cytokine cocktail of IL-1beta, IL-2 and IL-7 were added. After T cell priming for 7 days, wells were split and challenged for another 7 days with Ni or HEMA, and potentially cross-reactive haptens. Hapten-specificity of in vitro priming was demonstrated by proliferative responses to the haptens used for priming but not to the unrelated haptens. Highest priming efficiencies were obtained when both IL-4 and IL-12 were added to the cytokine supplement. Marked interferon-gamma (IFN-gamma) release (up to 4 ng/ml) was found when IL-12 was included in the cultures, whereas IL-5 release (up to 500 pg/ml) was observed after addition of IL-4 alone, or in combination with IL-12. Nickel-primed T cells showed frequent cross-reactivities with other metals closely positioned in the periodic table, i.e. palladium and copper, whereas HEMA-primed T cells showed distinct cross-reactivities with selected methacrylate congeners. Similar cross-reactivities are known to occur in allergic patients. Thus, in vitro T cell priming provides a promising tool for studying factors regulating cytokine synthesis, and cross-reactivity patterns of hapten-specific T cells.
authors.affiliation: Department of Pathology, Free University Hospital of Amsterdam, The Netherlands. t.rustemeijer@azvu.nl
authors.forename: T
authors.initials: T
authors.lastname: Rustemeyer
authors.forename: S
authors.initials: S
authors.lastname: De Ligter
authors.forename: B M
authors.initials: BM
authors.lastname: Von Blomberg
authors.forename: P J
authors.initials: PJ
authors.lastname: Frosch
authors.forename: R J
authors.initials: RJ
authors.lastname: Scheper
mesh_terms: D002478:Cells, Cultured; D003300:Copper / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D006241:Haptens / Q000276:immunology* / Q000494:pharmacology; D006801:Humans; D007114:Immunization; D018664:Interleukin-12 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D017493:Leukocyte Common Antigens / Q000096:biosynthesis; D008213:Lymphocyte Activation / Q000276:immunology*; D008689:Methacrylates / Q000494:pharmacology; D009532:Nickel / Q000276:immunology; D010165:Palladium / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
chemical_list: D016207:Cytokines; D018984:Epitopes, T-Lymphocyte; D006241:Haptens; D008689:Methacrylates; D018664:Interleukin-12; D015847:Interleukin-4; D010165:Palladium; C005044:hydroxyethyl methacrylate; D003300:Copper; D009532:Nickel; D017493:Leukocyte Common Antigens