title: Optimization of a dendritic cell-based assay for the in vitro priming of naïve human CD4+ T cells.
abstract: Methods to prime human CD4(+) T cells in vitro would be of significant value for the pre-clinical evaluation of vaccine candidates and other immunotherapeutics. However, to date, there is no reliable method for the induction of primary human T cell responses in the laboratory. Here, we optimized a culture strategy incorporating highly purified lymphocytes and dendritic cells, in the absence of any exogenous growth factors, for the in vitro sensitization of naïve CD4(+) T cells against a variety of protein antigens. This fully autologous approach, which was superior to the more traditional PBMC assay for supporting the induction of primary human T helper cell responses in culture, elicited effector cells capable of producing a variety of Th cytokines, including IFNgamma, TNFalpha, IL-2, IL-5, IL-17 and IL-21, and memory cells that could be restimulated multiple times with a specific antigen. Through simple modifications to this culture method, we evaluated the role of dendritic cell maturation state and regulatory T cells on the sensitization of naïve T helper cells, which highlights its utility for addressing basic questions of human immunobiology. Finally, using the formulated yellow fever vaccine, YF-VAX (R), we provide a proof-of-concept demonstration of the utility of the system for evaluating the T cell immunogenicity of vaccine candidates in a pre-clinical setting.
authors.affiliation: VaxDesign Corporation, 12612 Challenger Parkway, suite 365, Orlando, FL 32826, USA.
authors.forename: Janice M
authors.initials: JM
authors.lastname: Moser
authors.forename: Emily R
authors.initials: ER
authors.lastname: Sassano
authors.forename: Del C
authors.initials: del C
authors.lastname: Leistritz
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Eatrides
authors.forename: Sanjay
authors.initials: S
authors.lastname: Phogat
authors.forename: Wayne
authors.initials: W
authors.lastname: Koff
authors.forename: Donald R
authors.initials: DR
authors.lastname: Drake
mesh_terms: D017951:Antigen Presentation; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002469:Cell Separation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007156:Immunologic Memory; D007371:Interferon-gamma / Q000378:metabolism; D007378:Interleukins / Q000378:metabolism; D008213:Lymphocyte Activation; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D022223:Vaccines, Subunit / Q000276:immunology; D022341:Yellow Fever Vaccine / Q000276:immunology
chemical_list: D007378:Interleukins; D014409:Tumor Necrosis Factor-alpha; D022223:Vaccines, Subunit; D022341:Yellow Fever Vaccine; D007371:Interferon-gamma