FIG. 8.

CD11c⁺ DX5⁺ DC are the major producers of IFN-γ. (A) CD11c^hi DC subsets from naïve mice and mice infected with a low dose for 5 h were sorted on the basis of CD4 and CD8 expression, and ex vivo IFN-γ mRNA accumulation in sorted CD4⁺, CD8⁺, and DN DC was examined by real-time RT-PCR. Target genes were normalized against HPRT. Data represent the means plus standard errors of the means (error bars) from three separate experiments. (B) Splenic CD11c^hi cells from mice infected for 5 h with L. donovani were enriched using CD11c microbeads. Enriched CD11c^hi cells were then cultured in brefeldin A (10 μg/ml) for 4 h and surface stained with antibodies for CD11c, CD4, CD8, and CD49b (DX5). Fixed-cell samples were then permeabilized and stained for intracellular IFN-γ. Dot plots were gated on CD11c^hi cells. Quadrant gates were set on isotype controls. The numbers indicate the percentage of events within the three quadrants. This figure shows the results of one experiment that was representative of three independent experiments.