CCR1-Mediated STAT3 Tyrosine Phosphorylation and CXCL8 Expression in THP-1 Macrophage-like Cells Involve Pertussis Toxin-Insensitive Gα14/16 Signaling and IL-6 Release
CCR1-Mediated STAT3 Tyrosine Phosphorylation and CXCL8 Expression in THP-1 Macrophage-like Cells Involve Pertussis Toxin-Insensitive Gα14/16 Signaling and IL-6 Release
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DOI：
10.4049/jimmunol.1103359
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发表时间：
2012-12-01
影响因子：
4.4
通讯作者：
Wong, Yung H.
中科院分区：
文献类型：
--
作者：
Lee, Maggie M. K.;Chui, Ricky K. S.;Wong, Yung H.
CCR 1激动剂可能通过调节转录因子STAT 3促进超敏反应和动脉粥样硬化病变。CCR 1被证明使用百日咳毒素不敏感的G α（14/16）来刺激磷脂酶C β和NF-κ B，而G α（14）和G α（16）也能够激活STAT 3。CCR 1和G α（14/16）在人THP-1巨噬细胞样细胞中的共表达表明CCR 1可能利用G α（14/16）诱导STAT 3活化。在这项研究中，我们证明了CCR 1激动剂leukotactin-1（CCL 15）确实可以通过PMA分化的THP-1细胞、人红白血病细胞和过表达CCR 1和G α（14/16）的HEK 293细胞中百日咳毒素不敏感的G蛋白刺激STAT 3 Tyr（705）和Ser（727）磷酸化。STAT 3 Tyr（705）和Ser（727）磷酸化彼此独立并且在时间上不同。亚细胞分级分离和共聚焦显微镜显示，Tyr（705）-磷酸化的STAT 3易位到细胞核，而Ser（727）-磷酸化的STAT 3在CCR 1/G α（14）激活后保留在胞质溶胶中。CCL 15能够诱导THP-1巨噬细胞样细胞和过表达CCR 1和G α的HEK 293细胞产生IL-6和IL-8（CXCL 8）（14/16）。针对IL-6的中和Ab抑制了CCL 15介导的STAT 3 Tyr（705）磷酸化，而STAT 3活性的抑制消除了CCL 15激活的CXCL 8释放。CCR 1通过G α（14/16）传递信号的能力为CCL 15调节IL-6/STAT 3信号级联提供了联系，导致CXCL 8的表达，CXCL 8是一种参与炎症和动脉粥样硬化斑块破裂的细胞因子。免疫学杂志，2012，189：5266-5276。
Agonists of CCR1 contribute to hypersensitivity reactions and atherosclerotic lesions, possibly via the regulation of the transcription factor STAT3. CCR1 was demonstrated to use pertussis toxin-insensitive G alpha(14/16) to stimulate phospholipase C beta and NF-kappa B, whereas both G alpha(14) and G alpha(16) are also capable of activating STAT3. The coexpression of CCR1 and G alpha(14/16) in human THP-1 macrophage-like cells suggests that CCR1 may use G alpha(14/16) to induce STAT3 activation. In this study, we demonstrated that a CCR1 agonist, leukotactin-1 (CCL15), could indeed stimulate STAT3 Tyr(705) and Ser(727) phosphorylation via pertussis toxin-insensitive G proteins in PMA-differentiated THP-1 cells, human erythroleukemia cells, and HEK293 cells overexpressing CCR1 and G alpha(14/16). The STAT3 Tyr(705) and Ser(727) phosphorylations were independent of each other and temporally distinct. Subcellular fractionation and confocal microscopy illustrated that Tyr(705)-phosphorylated STAT3 translocated to the nucleus, whereas Ser(727)-phosphorylated STAT3 was retained in the cytosol after CCR1/G alpha(14) activation. CCL15 was capable of inducing IL-6 and IL-8 (CXCL8) production in both THP-1 macrophage-like cells and HEK293 cells overexpressing CCR1 and G alpha(14/16). Neutralizing Ab to IL-6 inhibited CCL15-mediated STAT3 Tyr(705) phosphorylation, whereas inhibition of STAT3 activity abolished CCL15-activated CXCL8 release. The ability of CCR1 to signal through G alpha(14/16) provides a linkage for CCL15 to regulate IL-6/STAT3-signaling cascades, leading to expression of CXCL8, a cytokine that is involved in inflammation and the rupture of atherosclerotic plaque. The Journal of Immunology, 2012, 189: 5266-5276.