A subset of human monocyte-derived dendritic cells expresses high levels of interleukin-12 in response to combined CD40 ligand and interferon-gamma treatment.
Dendritic cells (DCs) may arise from multiple lineages and progress through a series of intermediate stages until fully mature, at which time they are capable of optimal antigen presentation and T-cell activation. High cell surface expression of CD83 is presumed to correlate with full maturation of DCs, and a number of agents have been shown to increase CD83 expression on DCs. We hypothesized that interleukin 12 (IL-12) expression would be a more accurate marker of functionally mature DCs capable of activating antigen-specific T cells. We used combinations of signaling through CD40, using CD40 ligand trimer (CD40L), and interferon gamma to demonstrate that CD83 expression is necessary but not sufficient for optimal production of IL-12 by DCs. Phenotypically mature DCs could be induced to produce high levels of IL-12 p70 only when provided 2 simultaneous stimulatory signals. By intracellular cytokine detection, we determined that only a subset of cells that express high levels of CD80 and CD83 generate large amounts of IL-12. DCs matured with both signals are superior to DCs stimulated with the individual agents in activating antigen-specific T cell in vitro. These findings have important implications regarding the identification, characterization, and clinical application of functionally mature DCs.
Mosca
P J
PJ
Departments of General and Thoracic Surgery, Pathology, Immunology, and Internal Medicine, Center for Genetic and Cellular Therapies, Duke University Medical Center, Durham, NC, USA.
Hobeika
A C
AC
Clay
T M
TM
Nair
S K
SK
Thomas
E K
EK
Morse
M A
MA
Lyerly
H K
HK
D000276:Adjuvants, Immunologic / Q000378:metabolism; D017951:Antigen Presentation / Q000187:drug effects; D015703:Antigens, CD; D018122:B7-1 Antigen / Q000378:metabolism / Q000502:physiology; D015415:Biomarkers; D023201:CD40 Ligand / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003601:Cytotoxicity Tests, Immunologic; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism / Q000502:physiology; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000494:pharmacology*; D018664:Interleukin-12 / Q000276:immunology / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000502:physiology; D009000:Monocytes / Q000166:cytology; D021122:Protein Subunits; D000097802:CD83 Antigen
D000276:Adjuvants, Immunologic; D015703:Antigens, CD; D018122:B7-1 Antigen; D015415:Biomarkers; D023201:CD40 Ligand; D007136:Immunoglobulins; D007371:Interferon-gamma; D018664:Interleukin-12; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D021122:Protein Subunits; D000097802:CD83 Antigen