10.6084/m9.figshare.5125732.v1 Ayala A. Ayala A. Elphick G.F. Elphick G.F. Kim Y.S. Kim Y.S. Huang X. Huang X. Carreira-Rosario A. Carreira-Rosario A. Santos S.C. Santos S.C. Shubin N.J. Shubin N.J. Chen Y. Chen Y. Reichner J. Reichner J. Chung C.-S. Chung C.-S. Supplementary Material for: Sepsis-Induced Potentiation of Peritoneal Macrophage Migration Is Mitigated by Programmed Cell Death Receptor-1 Gene Deficiency Karger Publishers 2013 Macrophages Sepsis Co-inhibitory molecule Signal transduction Migration Phagocytosis 2013-11-16 00:00:00 Dataset https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Sepsis-Induced_Potentiation_of_Peritoneal_Macrophage_Migration_Is_Mitigated_by_Programmed_Cell_Death_Receptor-1_Gene_Deficiency/5125732 The effect of programmed cell death receptor-1 (PD-1) on phagocyte function has not been extensively described. Here we report that experimental mouse sepsis, cecal ligation and puncture (CLP), induced a marked increase in peritoneal macrophage random migration, motility and cell spread, but these changes were lost in the absence of PD-1. Alternatively, phagocytic activity was inversely affected. In vitro cell culture imaging studies, with the macrophage cell line J774, documented that blocking PD-1 with antibody led to aggregation of the cytoskeletal proteins α-actinin and F-actin. Further experiments looking at ex vivo peritoneal macrophages from mice illustrated that a similar pattern of α-actinin and F-actin was evident on cells from wild-type CLP mice but not PD-1-/- CLP mouse cells. We also observed that fMLP-induced migration by J774 cells was markedly attenuated using PD-1 blocking antibodies, a nonselective phosphatase inhibitor and a selective Ras-related protein 1 inhibitor. Finally, peritoneal macrophages derived from CLP as opposed to Sham mice demonstrated aspects of both cell surface co-localization with CD11b and internalization of PD-1 within vacuoles independent of CD11b staining. Together, we believe the data support a role for PD-1 in mediating aspects of innate macrophage immune dysfunction during sepsis, heretofore unappreciated.