Pittsburgh Center for Pain Research, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States; Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States; Center for Neuroscience, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States.
Pittsburgh Center for Pain Research, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States; Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States.
Brain Behav Immun. 2022 Nov;106:233-246. doi: 10.1016/j.bbi.2022.09.001. Epub 2022 Sep 8.
PDL1 is a protein that induces immunosuppression by binding to PD1 expressed on immune cells. In line with historical studies, we found that membrane-bound PD1 expression was largely restricted to immune cells; PD1 was not detectable at either the mRNA or protein level in peripheral neurons using single neuron qPCR, immunolabeling and flow cytometry. However, we observed widespread expression of PDL1 in both sensory and sympathetic neurons that could have important implications for patients receiving immunotherapies targeting this pathway that include unexpected autonomic and sensory related effects. While signaling pathways downstream of PD1 are well established, little to no information is available regarding the intracellular signaling downstream of membrane-bound PDL1 (also known as reverse signaling). Here, we administered soluble PD1 to engage neuronally expressed PDL1 and found that PD1 significantly reduced nocifensive behaviors evoked by algogenic capsaicin. We used calcium imaging to examine the underlying neural mechanism of this reduction and found that exogenous PD1 diminished TRPV1-dependent calcium transients in dissociated sensory neurons. Furthermore, we observed a reduction in membrane expression of TRPV1 following administration of PD1. Exogenous PD1 had no effect on pain-related behaviors in sensory neuron specific PDL1 knockout mice. These data indicate that neuronal PDL1 activation is sufficient to modulate sensitivity to noxious stimuli and as such, may be an important homeostatic mechanism for regulating acute nociception.
PDL1 是一种蛋白,通过与免疫细胞表达的 PD1 结合来诱导免疫抑制。与历史研究一致,我们发现膜结合 PD1 的表达主要局限于免疫细胞;使用单细胞 qPCR、免疫标记和流式细胞术,在外周神经元中既不能在 mRNA 水平也不能在蛋白水平检测到 PD1。然而,我们观察到在感觉神经元和交感神经元中广泛表达 PDL1,这可能对接受针对该途径的免疫治疗的患者具有重要意义,因为这些免疫治疗会产生意料之外的自主和感觉相关的影响。虽然 PD1 下游的信号通路已经得到很好的研究,但关于膜结合 PDL1(也称为反向信号)下游的细胞内信号通路的信息却很少。在这里,我们给予可溶性 PD1 以结合神经元表达的 PDL1,发现 PD1 显著减少了致痛性辣椒素诱发的伤害性行为。我们使用钙成像来研究这种减少的潜在神经机制,发现外源性 PD1 减弱了分离感觉神经元中 TRPV1 依赖性钙瞬变。此外,我们观察到 PD1 给药后 TRPV1 的膜表达减少。外源性 PD1 对感觉神经元特异性 PDL1 敲除小鼠的疼痛相关行为没有影响。这些数据表明神经元 PDL1 的激活足以调节对有害刺激的敏感性,因此可能是调节急性痛觉的重要体内平衡机制。