Suppressors of Cytokine Signaling (SOCS)1 and SOCS3 Proteins Are Mediators of Interleukin-10 Modulation of Inflammatory Responses Induced by Chlamydia muridarum and Its Major Outer Membrane Protein (MOMP) in Mouse J774 Macrophages

Figure 6

Skewing of the chlamydial M1 pro-inflammatory phenotype to an M2 anti-inflammatory phenotype by exogenous and endogenously produced IL-10. Macrophages (10⁶ cells/mL) were stimulated with rMOMP (0.1, 1 and 10 μg/mL) (A-C) or infected with Cm (MOI of 0.5, 1 and 2) (D-F) in the presence and absence of IL-10 (10 ng/mL). Macrophages were stimulated with rMOMP (1 μg/mL; G-I) or Cm (MOI of 2; J-L) in the presence and absence of IL-10 (0.1, 1, and 10 ng/mL). At 24 h post-stimulation, the mRNA transcripts of the m1 marker; nos2 and m2 markers; arg1 and mrc1 were quantified using TaqMan qRT-PCR. The m1/m2 (M) and m2/m1 (N) ratios were calculated from macrophages exposed to rMOMP or Cm in the presence and absence of IL-10. Protein expressions of nos2 and mrc1 were evaluated from stimulated cells employing flow cytometry (O-T). Macrophages pre-incubated with neutralizing Abs to IL-10, IL-6, and TNF were stimulated with rMOMP or Cm for an additional 24 h. Normal rat IgG1 Ab served as the isotype control (ISO). Post-stimulation, RNA was isolated to quantify the gene transcripts of nos2 and mrc1 (U-X) by TaqMan qRT-PCR. An asterisk indicates significant differences (P <0.05), and P values were calculated as described in Figure 1. Each bar represents the mean ± SD of samples run in triplicates, and each experiment was repeated at least 3 to 4 times with a representative experiment shown.

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