Programmed death-ligand 1 (PD-L1) Modulates Chemokine Production Via the TLR4/TRAF6 Signaling Axis During LPS + IFN-γ-Induced Endotoxemia-mimicked Sepsis.

As a pivotal immune checkpoint molecule, programmed death-ligand 1 (PD-L1) is anchored primarily on the membrane surface of immune cells, where it exerts immunosuppressive effects, thereby facilitating tumor immune evasion. Macrophages, which serve as essential sentinels of the innate immune system, play dual regulatory roles in inflammatory pathologies, particularly during sepsis progression. While their secreted chemokines mediate inflammatory cell recruitment for pathogen clearance, excessive chemokine production can paradoxically induce organ damage and immune cell exhaustion, necessitating precise regulatory mechanisms. Conventional understanding suggests that PD-L1 on macrophages engages with programmed cell death protein 1 (PD-1) on T lymphocytes to suppress T-cell proliferation, cytokine secretion (e.g., IFN-γ and IL-2), and cytotoxic functions, thereby negatively modulating adaptive immunity. However, emerging evidence also suggests that PD-L1 has context-dependent proinflammatory functions. Given this context, we hypothesized that macrophage-intrinsic PD-L1 plays a poorly understood role in directly regulating chemokine production during sepsis. Our integrative analysis incorporating clinical database mining and RNA sequencing (RNA-seq) revealed a less defined proinflammatory property of PD-L1 under septic conditions-an ability to increase CCL8 and CXCL9 chemokine expression in inflammatory macrophages. Through combinatorial approaches, including immunoprecipitation‒mass spectrometry (IP‒MS), molecular docking, and site-directed mutagenesis, we preliminarily elucidated that PD-L1 likely governs the chemotactic mediators CCL8 and CXCL9 via the TLR4/TRAF6 signaling axis. These findings collectively establish the previously unappreciated regulatory capacity of macrophage-intrinsic PD-L1 in chemokine modulation during sepsis, potentially informing the development of innovative therapeutic strategies targeting immune dysregulation in critical care settings.