Enhanced T-cell activation and T-cell-dependent IL-2 production by CD83+, CD25high, CD43high human monocyte-derived dendritic cells.
Although standardized protocols are widely used for the generation of monocyte-derived immunostimulatory dendritic cells (DC(ims)), the inducibility of Th1 cells by DC(ims) may considerably differ. As a measure for the quality of DC(ims) generated from an individual donor at a certain time point, CD83 is used in combination with HLA-DR and CD86 to assess DC maturation. When phenotypically analyzing DC(ims), we identified a subpopulation ( approximately 60%) of CD83+, CD86+, and HLA-DR+ DC(ims) that co-expressed CD25. DC within a given DC(ims) preparation identified by lower expression of CD83 and by selective expression of CD14, however, did not co-express CD25. In order to establish CD25 as an additional maturation marker of DC(ims), we studied the DC phenotype of these cells as well as the DC-dependent T-cell proliferation and T-cell cytokine production profile after co-incubation with sorted CD25(high) and CD25(low) subpopulations of CD83+, HLA-DR+, CD86+ DC(ims). CD25(high) DC(ims) showed significant up-regulation of the DC activation molecule CD43 and induced increased levels of IL-2 secretion in allogeneic T-cells (170.7+/-86.7pg/mL) as compared to T-cells coincubated with CD25(low) DC(ims) (86.6+/-37.6pg/mL) [p=0.0224]. This was reflected by a significantly lower T-cell stimulatory capacity of CD25(low) DC(ims) (84.0% of CD25(high) DC(ims), 1:10 ratio; p=0.014) whereas the T-cell stimulatory capacity of CD25(low) DC(ims) was much higher when compared to IL-10 induced regulatory DC (55.3% of CD25(high) DC(ims); 1:10 ratio). With regard to cancer vaccination protocols, we propose to use CD25 and CD43 as additional markers for DC quality control, assessment of maturational status, and positive selection.
Velten
Florian W
FW
Department of Dermatology, University Medical Center Mannheim, Ruprecht Karls University of Heidelberg, Theodor-Kutzer Ufer 1-3, 68167 Mannheim, Germany. Florian.velten@haut.ma.uni-heidelberg.de
Rambow
Florian
F
Metharom
P
P
Goerdt
Sergij
S
D015703:Antigens, CD / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000737:chemistry / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D007376:Interleukin-2 / Q000378:metabolism*; D053645:Interleukin-2 Receptor alpha Subunit / Q000032:analysis / Q000378:metabolism; D051917:Leukosialin / Q000032:analysis; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000032:analysis; D009000:Monocytes / Q000737:chemistry / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology*; D000097802:CD83 Antigen
D015703:Antigens, CD; D015415:Biomarkers; D016207:Cytokines; D007136:Immunoglobulins; D007376:Interleukin-2; D053645:Interleukin-2 Receptor alpha Subunit; D051917:Leukosialin; D008562:Membrane Glycoproteins; D000097802:CD83 Antigen