Phenotypic characterization of DC subsets. (A,B) DCs were propagated from bone marrow precursors under the conditions indicated with or without IL-10 in FBS- or NMS-supplemented cultures. On day 8, cells were harvested and analyzed for surface markers either unstimulated (A) or stimulated with 1 μg/mL LPS for the last 24 hours (B). Histograms shown are representative of 3 to 7 observations. (C) DCs were pulsed or not with 10 μg/mL NP-118 peptide for the last 24 hours of culture and used as APCs for splenocytes from LCMV-infected syngeneic mice and analyzed by intracellular cytokine staining (ICCS) as described. As positive control, peptide was added directly to splenocytes or splenocytes were left unstimulated. (D) Phenotype of NMS DCs grown with or without IL-4. (E) DCs were used as stimulator cells in an allogeneic mixed lymphocyte reaction (MLR) using 105 C57BL/6-derived T cells as responders. T-cell activation was measured by 3H-thymidine incorporation after 5 days.
