FIGURE 3.

High levels of IL-10 production in the culture supernatants of PBMCs from patients with uncomplicated malaria in the presence of IL-2 and Pf Ag for 10 days. Fold increase in the number of γδ T cell subsets (A) and the production of IL-10 and IFN-γ (B) after the culture of PBMCs from NECs, Japanese healthy controls (JHCs), negative controls (NCs), and non-hospitalized uncomplicated malaria patients (UMPs). PBMCs (1–2 × 10⁶ cells/mL) were cultured in medium containing 10% fetal calf serum with or without IL-2 (50 U/mL) or IL-2/Pf Ag (5 μg/mL) for 10 days. Cytokines in the culture supernatants were detected with a CBA Flex Set assay. Flow cytometric profiles of intracellular IL-10 and IFN-γ staining in γδ T cells (C), and the numbers of IL-10-producing (D, left) and IFN-γ-producing cells (D, right) in the total PBMCs and γδ T cell subsets per 10⁶ PBMCs from 3–5 JHCs and 3–5 UMPs after culture for 10 days. Three-color staining was performed with mAbs directed against γδTCR, Vγ9TCR, IL-10, and IFN-γ. The numbers in the figures represent the mean percentages of immunofluorescence-positive cells. Representative results of three experiments are shown. Statistical analyses were performed with one-way ANOVA with Dunnett’s post hoc test (B) and unpaired t-test (D). ^∗p < 0.05, ^∗∗p < 0.01.