title: Potency of mature CD40L RNA electroporated dendritic cells correlates with IL-12 secretion by tracking multifunctional CD8(+)/CD28(+) cytotoxic T-cell responses in vitro.
abstract: Electroporation of mature dendritic cells (DC) with RNA-encoding CD40L greatly enhances the production of interleukin (IL)-12, a proinflammatory cytokine necessary for the induction of T-cell immunity. Results presented herein reveal a correlation between the priming of CD28(+) antigen-reactive effector memory cytotoxic T lymphocytes (CTL) displaying 3 or 4 simultaneous effector functions and the quantity of IL-12 produced by postmaturation electroporation-CD40L DC. By using multiparameter flow cytometry, the quantities of IL-12 needed to prime naive antigen-reactive T cells to simultaneously produce interferon-γ and tumor necrosis factor-α in the presence or absence of IL-2 secretion in conjunction with lytic activity defined by CD107a expression can be used to determine the overall potency of a DC product. In the presence of IL-12, CTL differentiation toward lytic function is not accompanied by a reduction in the secretion of interferon-γ and tumor necrosis factor-α. Therefore, by measuring the availability of IL-12 one can predict the potency of a DC immunotherapeutics in relation to its ability to drive distinct effector memory CTL subsets with multifunctional activities.
authors.affiliation: Research Department, Argos Therapeutics, Inc, Durham, NC 27704, USA. mdebenedette@argostherapeutics.com
authors.forename: Mark A
authors.initials: MA
authors.lastname: DeBenedette
authors.forename: David M
authors.initials: DM
authors.lastname: Calderhead
authors.forename: Irina Y
authors.initials: IY
authors.lastname: Tcherepanova
authors.forename: Charles A
authors.initials: CA
authors.lastname: Nicolette
authors.forename: Don G
authors.initials: DG
authors.lastname: Healey
mesh_terms: D018106:CD28 Antigens / Q000096:biosynthesis; D023201:CD40 Ligand / Q000235:genetics*; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism*; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D018664:Interleukin-12 / Q000096:biosynthesis / Q000378:metabolism*; D007376:Interleukin-2 / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D052118:Lysosomal-Associated Membrane Protein 1 / Q000235:genetics; D012313:RNA / Q000235:genetics*; D015398:Signal Transduction; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D018106:CD28 Antigens; D007376:Interleukin-2; D052118:Lysosomal-Associated Membrane Protein 1; D014409:Tumor Necrosis Factor-alpha; D023201:CD40 Ligand; D018664:Interleukin-12; D012313:RNA; D007371:Interferon-gamma