Figure 4.

Endogenously produced IL-10 regulates inflammatory mediators and SOCS expression in chlamydial-stimulated macrophages. Macrophages (10⁶/mL) were stimulated for 24 h with rMOMP (10 μg/mL) or Cm (MOI of 2) to quantify the production of endogenously produced IL-10 in supernatants by ELISA (A). Macrophages were pre-incubated with neutralizing Ab (αIL-10) to IL-10 (25 μg/mL) for 30 min before adding rMOMP (10 μg/mL) or Cm (MOI of 2) for an additional 24 h. Normal rat IgG1 Ab served as the isotype control (ISO). Post-stimulation, supernatants were collected to quantify IL-6 (B, F), IL-12p40 (C), TNF (D), and CCL5 (E, G) by specific ELISAs and RNA was isolated to quantify the mRNA gene transcripts of SOCS1 and SOCS3 (H-I) by TaqMan qRT-PCR. An asterisk indicates a significant difference (P <0.05), and P values were calculated as described in Figure 1 legend. Each bar represents the mean ± SD of samples run in triplicates, and each experiment was repeated at least 3 times.