Figure 2
Figure 2. PDCs selectively expand T cells that produce cytokines on rechallenge with antigen. Cocultures of CFSE-labeled CD4+ T cells and PDCs or CD11c+ DCs were stimulated with HCMV in the absence or presence of neutralizing anti–IL-10 or anti–type I IFN, as indicated. On day 6, expanded T cells were restimulated for 16 hours with autologous imoDCs and HCMV. The production of IL-10 and IFN-γ was measured in CD3+ T cells with the cytokine secretion assay and was almost exclusively found in the CD3+ T cells that had divided (CFSElow), as shown for IL-10 in the upper plots of panel A. The lower 6 plots are gated on dividing cells, as indicated, and show the secretion of IL-10 and IFN-γ in one experiment representative of 4 performed. Percentages of single and double cytokine-producing cells are indicated. In panel B the results from 4 different experiments are shown, with means and SEM indicated. All cytokine-producing populations were higher in PDC-expanded as compared with CD11c+ DC-expanded cultures (*P < .05). Addition of neutralizing anti–IL-10 resulted in significantly higher levels of double cytokine-producing cells in the PDC cocultures (**P < .02), whereas the levels were nonsignificantly changed in the presence of anti–type I IFN. (C) Blocking of IL-12, IL-18, or IL-1β did not reduce the levels of IL-10 produced in PDC cocultures relative to control (stippled line), as measured in the culture supernatants at 48 hours following stimulation with HCMV. Two separate experiments are indicated by distinct symbols.

PDCs selectively expand T cells that produce cytokines on rechallenge with antigen. Cocultures of CFSE-labeled CD4+ T cells and PDCs or CD11c+ DCs were stimulated with HCMV in the absence or presence of neutralizing anti–IL-10 or anti–type I IFN, as indicated. On day 6, expanded T cells were restimulated for 16 hours with autologous imoDCs and HCMV. The production of IL-10 and IFN-γ was measured in CD3+ T cells with the cytokine secretion assay and was almost exclusively found in the CD3+ T cells that had divided (CFSElow), as shown for IL-10 in the upper plots of panel A. The lower 6 plots are gated on dividing cells, as indicated, and show the secretion of IL-10 and IFN-γ in one experiment representative of 4 performed. Percentages of single and double cytokine-producing cells are indicated. In panel B the results from 4 different experiments are shown, with means and SEM indicated. All cytokine-producing populations were higher in PDC-expanded as compared with CD11c+ DC-expanded cultures (*P < .05). Addition of neutralizing anti–IL-10 resulted in significantly higher levels of double cytokine-producing cells in the PDC cocultures (**P < .02), whereas the levels were nonsignificantly changed in the presence of anti–type I IFN. (C) Blocking of IL-12, IL-18, or IL-1β did not reduce the levels of IL-10 produced in PDC cocultures relative to control (stippled line), as measured in the culture supernatants at 48 hours following stimulation with HCMV. Two separate experiments are indicated by distinct symbols.

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