The vimentin rod domain blocks P-selectin-P-selectin glycoprotein ligand 1 interactions to attenuate leukocyte adhesion to inflamed endothelium

• Published in: PloS one Vol. 15; no. 10; p. e0240164
• Main Authors: Lam, Fong Wilson, Brown, Cameron August, Valladolid, Christian, Emebo, Dabel Cynthia, Palzkill, Timothy Gerald, Cruz, Miguel Angel
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 13.10.2020; Public Library of Science (PLoS)
• Subjects: Adhesion; Animals; Antibodies; Arrays; Attenuation; Binding sites; Biology and Life Sciences; Blocking; CD162 antigen; Cell adhesion; Cell Adhesion - drug effects; Cell Adhesion - immunology; Cellulose; Cloning; Cytokines; Disease Models, Animal; Domains; E coli; Endothelial cells; Endothelium; Endothelium - drug effects; Endothelium - immunology; Endotoxemia - drug therapy; Endotoxemia - immunology; Endotoxemia - pathology; Female; Fibrin; Funding; Glycoproteins; Health aspects; Healthy Volunteers; Human Umbilical Vein Endothelial Cells; Humans; IL-1β; Inflammation; Inflammatory diseases; Interferometry; Interleukin 4; Intermediate filament proteins; Leukocytes; Leukocytes - drug effects; Leukocytes - immunology; Ligands; Ligands (Biochemistry); Liver; Male; Medical schools; Medicine; Medicine and Health Sciences; Membrane Glycoproteins - metabolism; Membranes; Molecular Docking Simulation; P-selectin; P-Selectin - metabolism; P-selectin glycoprotein ligand 1; Pediatrics; Peptides; Physiological aspects; Platelets; Protein Binding - drug effects; Protein Binding - immunology; Protein Domains - genetics; Protein expression; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - pharmacology; Recombinant Proteins - therapeutic use; Research and Analysis Methods; Sepsis; Sine waves; Umbilical vein; Veterans; Vimentin; Vimentin - genetics; Vimentin - pharmacology; Vimentin - therapeutic use; White blood cells; United States; United States > US; Texas; Germany
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0240164

Acute inflammation begins with leukocyte P-selectin glycoprotein ligand-1 (PSGL-1) binding to P-selectin on inflamed endothelium and platelets. In pathologic conditions, this process may contribute to secondary organ damage, like sepsis-induced liver injury. Therefore, developing novel therapies to attenuate inflammation may be beneficial. We previously reported that recombinant human vimentin (rhVim) binds P-selectin to block leukocyte adhesion to endothelium and platelets. In this study, we used SPOT-peptide arrays to identify the rod domain as the active region within rhVim that interacts with P-selectin. Indeed, recombinant human rod domain of vimentin (rhRod) binds to P-selectin with high affinity, with in silico modeling suggesting that rhRod binds P-selectin at or near the PSGL-1 binding site. Using bio-layer interferometry, rhRod decreases PSGL-1 binding to immobilized P-selectin, corroborating the in silico data. Under parallel-plate flow, rhRod blocks leukocyte adhesion to fibrin(ogen)-captured platelets, P-selectin/Fc-coated channels, and IL-1β/IL-4-co-stimulated human umbilical vein endothelial cells. Finally, using intravital microscopy in endotoxemic C57Bl/6 mice, rhRod co-localizes with P-selectin in the hepatic sinusoids and decreases neutrophil adhesion to hepatic sinusoids. These data suggest a potential role for rhRod in attenuating inflammation through directly blocking P-selectin-PSGL-1 interactions.