The immunopathology of chlamydial diseases is exacerbated by a broad-spectrum of inflammatory mediators, which we reported are inhibited by IL-10 in macrophages. However, the chlamydial protein moiety that induces the inflammatory mediators and the mechanisms by which IL-10 inhibits them are unknown. We hypothesized that Chlamydia major outer membrane protein (MOMP) mediates its disease pathogenesis, and the suppressor of cytokine signaling (SOCS)1 and SOCS3 proteins are mediators of the IL-10 inhibitory actions. Our hypothesis was tested by exposing mouse J774 macrophages to chlamydial stimulants (live Chlamydia muridarum and MOMP) with and without IL-10. MOMP significantly induced several inflammatory mediators (IL-6, IL-12p40, CCL5, CXCL10), which were dose-dependently inhibited by IL-10. Chlamydial stimulants induced the mRNA gene transcripts and protein expression of SOCS1 and SOCS3, with more SOCS3 expression. Notably, IL-10 reciprocally regulated their expression by reducing SOCS1 and increasing SOCS3. Specific inhibitions of MAPK pathways revealed that p38, JNK, and MEK1/2 are required for inducing inflammatory mediators as well as SOCS1 and SOCS3. Chlamydial stimulants triggered an M1 pro-inflammatory phenotype evidently by an enhanced nos2 (M1 marker) expression, which was skewed by IL-10 towards a more M2 anti-inflammatory phenotype by the increased expression of mrc1 and arg1 (M2 markers) and the reduced SOCS1/SOCS3 ratios. Neutralization of endogenously produced IL-10 augmented the secretion of inflammatory mediators, reduced SOCS3 expression, and skewed the chlamydial M1 to an M2 phenotype. Inhibition of proteasome degradation increased TNF but decreased IL-10, CCL5, and CXCL10 secretion by suppressing SOCS1 and SOCS3 expressions and dysregulating their STAT1 and STAT3 transcription factors. Our data show that SOCS1 and SOCS3 are regulators of IL-10 inhibitory actions, and underscore SOCS proteins as therapeutic targets for IL-10 control of inflammation for Chlamydia and other bacterial inflammatory diseases.

中文翻译：

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细胞因子信号转导抑制因子 (SOCS)1 和 SOCS3 蛋白是 IL-10 调节小鼠 J774 巨噬细胞中由鼠衣原体及其主要外膜蛋白 (MOMP) 诱导的炎症反应的介质。

广谱炎症介质加剧了衣原体疾病的免疫病理学，我们报道过巨噬细胞中的 IL-10 会抑制这些介质。然而，诱导炎症介质的衣原体蛋白部分以及 IL-10 抑制它们的机制尚不清楚。我们假设衣原体主要外膜蛋白 (MOMP) 介导其疾病发病机制，细胞因子信号转导抑制因子 (SOCS)1 和 SOCS3 蛋白是 IL-10 抑制作用的介质。我们的假设通过将小鼠 J774 巨噬细胞暴露于衣原体兴奋剂（活鼠衣原体和 MOMP) 有和没有 IL-10。MOMP 显着诱导几种炎症介质（IL-6、IL-12p40、CCL5、CXCL10），这些介质受到 IL-10 的剂量依赖性抑制。衣原体兴奋剂诱导SOCS1和SOCS3的mRNA基因转录和蛋白表达，SOCS3表达较多。值得注意的是，IL-10 通过减少 SOCS1 和增加 SOCS3 相互调节它们的表达。对 MAPK 通路的特异性抑制表明，p38、JNK 和 MEK1/2 是诱导炎症介质以及 SOCS1 和 SOCS3 所必需的。衣原体兴奋剂显然通过增强的 nos2（M1 标记）表达触发了 M1 促炎表型，而 IL-10 通过 mrc1 和 arg1（M2 标记）的表达增加以及减少的SOCS1/SOCS3 比率。内源性产生的 IL-10 的中和增加了炎症介质的分泌，减少了 SOCS3 的表达，并使衣原体 M1 表型偏向 M2 表型。通过抑制 SOCS1 和 SOCS3 表达并失调其 STAT1 和 STAT3 转录因子，抑制蛋白酶体降解会增加 TNF，但会减少 IL-10、CCL5 和 CXCL10 的分泌。我们的数据显示 SOCS1 和 SOCS3 是 IL-10 抑制作用的调节剂，并强调 SOCS 蛋白是 IL-10 控制炎症的治疗靶点 通过抑制 SOCS1 和 SOCS3 的表达并调节其 STAT1 和 STAT3 转录因子来调节 CXCL10 和 CXCL10 的分泌。我们的数据显示 SOCS1 和 SOCS3 是 IL-10 抑制作用的调节剂，并强调 SOCS 蛋白是 IL-10 控制炎症的治疗靶点 通过抑制 SOCS1 和 SOCS3 的表达并调节其 STAT1 和 STAT3 转录因子来调节 CXCL10 和 CXCL10 的分泌。我们的数据显示 SOCS1 和 SOCS3 是 IL-10 抑制作用的调节剂，并强调 SOCS 蛋白是 IL-10 控制炎症的治疗靶点衣原体和其他细菌性炎症性疾病。