title: Efficient priming of protein antigen-specific human CD4(+) T cells by monocyte-derived dendritic cells.
abstract: Dendritic cells (DCs) have the unique ability to initiate an immune response in vivo by capturing antigens (Ags) in peripheral tissues and migrating to secondary lymphoid organs, where they sensitize naive CD4(+) T cells. To mimic this process in vitro, previous studies have shown that DCs directly isolated from peripheral blood can be used to elicit primary responses to neoantigens (neoAgs). In other studies, when monocyte-derived DCs have been utilized to sensitize total CD4(+) T cells in vitro, only secondary proliferation to neoAgs could be elicited. In the present study, the relative abilities of CD40 ligation, protein kinase C activation, and culture in tumor necrosis factor alpha (TNF-alpha) to induce functional and phenotypic maturation of human DCs from monocyte precursors were compared. Optimal TNF-alpha-induced maturation of DCs required a prolonged 4-day culture. It was then found that loading immature DCs with the neoAgs keyhole limpet hemocyanin or human immunodeficiency virus-1 p24 gag prior to TNF-alpha-induced maturation, rather than after maturation, was crucial to sensitize CD4(+) T cells to new Ags. This primary proliferation to neoAgs was initiated from the CD4(+) CD45RA(+) naive T-cell population. Finally, it was found that monocyte-derived DCs acquired the ability to secrete interleukin-12 p70, after contact with Ag-specific T cells. The ability to prime and expand Ag-specific CD4(+) T cells ex vivo to neoAgs in serum-free conditions has potential application for cellular vaccination and adoptive immunotherapy.
authors.affiliation: Department of Molecular and Cellular Engineering, University of Pennsylvania, Philadelphia, USA.
authors.forename: K
authors.initials: K
authors.lastname: Schlienger
authors.forename: N
authors.initials: N
authors.lastname: Craighead
authors.forename: K P
authors.initials: KP
authors.lastname: Lee
authors.forename: B L
authors.initials: BL
authors.lastname: Levine
authors.forename: C H
authors.initials: CH
authors.lastname: June
mesh_terms: D000276:Adjuvants, Immunologic / Q000096:biosynthesis; D017951:Antigen Presentation / Q000276:immunology*; D000938:Antigen-Presenting Cells / Q000166:cytology / Q000276:immunology; D000941:Antigens / Q000276:immunology / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D023201:CD40 Ligand / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D019070:Cell Lineage / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D006433:Hemocyanins / Q000276:immunology / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D018664:Interleukin-12 / Q000096:biosynthesis; D017493:Leukocyte Common Antigens / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D009000:Monocytes / Q000166:cytology; D013744:Tetanus Toxin / Q000276:immunology / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D000276:Adjuvants, Immunologic; D000941:Antigens; D013744:Tetanus Toxin; D014409:Tumor Necrosis Factor-alpha; D023201:CD40 Ligand; D018664:Interleukin-12; D006433:Hemocyanins; D017493:Leukocyte Common Antigens; C032808:keyhole-limpet hemocyanin; D013755:Tetradecanoylphorbol Acetate