Figure 5.

Induction of IL-10 production in monocytes by HRV-14. (a) Supernatants of monocytes (1 × 10⁶/mL) cultured in the presence of HRV-14 (10 TCID₅₀ per cell), LPS (100 ng/mL), or medium for 2 days were analyzed for IL-10, IL-1β, and TNF-α production by ELISA. The figure shows mean values ± SEM of 3 experiments. (b) In separate experiments, detection of cytokine production in monocytes was performed by cytoplasmic staining with specific mAb’s. Monocytes (1 × 10⁶/mL) were cultured for 24 hours in the presence of HRV-14 (10 TCID₅₀ per cell) or LPS (100 ng/mL). After 12 hours of culture, monensin (5 μM) was added in all instances. The cells were harvested, fixed, permeabilized, and subsequently stained with PE-labeled anti–IL-10 mAb. For determination of IL-1β or TNF-α production by cytoplasmic staining, monocytes were incubated with the indicated stimuli for 6 hours in the presence of monensin (5 μM). Cytoplasmic cytokine expression was analyzed by flow cytometry. Overlay histograms show expression in mock-treated cells (open histograms) and monocytes stimulated with HRV-14 (gray histograms) or LPS (open histograms, thick line). The results shown are representative of 3 experiments.