Graduate Program in Immunology, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, Massachusetts, USA.
PLoS One. 2010 Sep 22;5(9):e12871. doi: 10.1371/journal.pone.0012871.
Toll-like receptor (TLR)-2/TLR1 heterodimers recognize bacterial lipopeptides and initiate the production of inflammatory mediators. Adaptors and co-receptors that mediate this process, as well as the mechanisms by which these adaptors and co-receptors function, are still being discovered.
METHODOLOGY/PRINCIPAL FINDINGS: Using shRNA, blocking antibodies, and fluorescent microscopy, we show that U937 macrophage responses to the TLR2/1 ligand, Pam(3)CSK(4), are dependent upon an integrin, α(3)β(1). The mechanism for integrin α(3)β(1) involvement in TLR2/1 signaling is through its role in endocytosis of lipopeptides. Using inhibitors of endosomal acidification/maturation and physical tethering of the ligand, we show that the endocytosis of Pam(3)CSK(4) is necessary for the complete TLR2/1-mediated pro-inflammatory cytokine response. We also show that TLR2/1 signaling from the endosome results in the induction of different inflammatory mediators than TLR2/1 signaling from the plasma membrane.
CONCLUSION/SIGNIFICANCE: Here we identify integrin α(3)β(1) as a novel regulator for the recognition of bacterial lipopeptides. We demonstrate that induction of a specific subset of cytokines is dependent upon integrin α(3)β(1)-mediated endocytosis of the ligand. In addition, we address an ongoing controversy regarding endosomal recognition of bacterial lipopeptides by demonstrating that TLR2/1 signals from within endosomal compartments as well as the plasma membrane, and that downstream responses may differ depending upon receptor localization. We propose that the regulation of endosomal TLR2/1 signaling by integrin α(3)β(1) serves as a mechanism for modulating inflammatory responses.
Toll 样受体(TLR)-2/TLR1 异二聚体识别细菌脂肽并启动炎症介质的产生。介导此过程的衔接蛋白和共受体,以及这些衔接蛋白和共受体发挥作用的机制,仍在探索中。
方法/主要发现:使用 shRNA、阻断抗体和荧光显微镜,我们发现 U937 巨噬细胞对 TLR2/1 配体 Pam(3)CSK(4)的反应依赖于整合素α(3)β(1)。整合素α(3)β(1)参与 TLR2/1 信号转导的机制是通过其在脂肽内吞作用中的作用。通过使用内体酸化/成熟抑制剂和配体的物理束缚,我们表明 Pam(3)CSK(4)的内吞作用对于完整的 TLR2/1 介导的促炎细胞因子反应是必需的。我们还表明,内体中的 TLR2/1 信号转导导致诱导的炎症介质与质膜中的 TLR2/1 信号转导不同。
结论/意义:在这里,我们确定整合素α(3)β(1)是细菌脂肽识别的新型调节剂。我们证明,诱导特定细胞因子亚群的产生依赖于配体的整合素α(3)β(1)介导的内吞作用。此外,我们通过证明 TLR2/1 信号从内体区室以及质膜发出,并且下游反应可能取决于受体定位,解决了关于内体中细菌脂肽的 TLR2/1 识别的持续争议。我们提出,整合素α(3)β(1)对内体 TLR2/1 信号转导的调节可作为调节炎症反应的机制。