title: A novel microfluidic assay reveals a key role for protein kinase C δ in regulating human neutrophil-endothelium interaction.
abstract: A key step in neutrophil-mediated tissue damage is the migration of activated neutrophils across the vascular endothelium. Previously, we identified protein kinase C δ as a critical regulator of neutrophil migration in sepsis but did not identify specific steps in migration. In this study, we used our novel biomimetic microfluidic assay to delineate systematically the mechanism by which protein kinase C δ regulates individual steps in human neutrophil-endothelial interaction during inflammation. The biomimetic microfluidic assay includes a network of vascular channels, produced from in vivo images connected to a tissue compartment through a porous barrier. HUVECs cultured in vascular channels formed a complete lumen under physiologic shear flow. HUVECs were pretreated with TNF-α ± a protein kinase C δ inhibitor, and the tissue compartment was filled with a chemoattractant (fMLP or IL-8). Under physiologic shear flow, the role of protein kinase C δ on spatial and temporal neutrophil adherence/migration was quantified. Protein kinase C δ inhibition significantly reduced neutrophil adhesion in response to fMLP and IL-8 only under low shear rate and near bifurcations. Protein kinase C δ inhibition also decreased adherence to nonactivated HUVECs in response to fMLP or IL-8. Protein kinase C δ inhibition reduced neutrophil migration into the tissue compartment in response to fMLP and to a lesser degree, to IL-8. Antibody-coated microparticles demonstrated that protein kinase C δ inhibition down-regulated E-selectin and ICAM-1 but not VCAM-1 expression. With the use of a physiologically relevant in vitro model system, we demonstrate that protein kinase C δ plays an important role in the regulation of neutrophil adherence/migration during inflammation and identifies key steps regulated by protein kinase C δ in neutrophil-endothelial interactions.
authors.affiliation: Department of Mechanical Engineering, College of Engineering, Temple University, Philadelphia, Pennsylvania, USA.
authors.forename: Fariborz
authors.initials: F
authors.lastname: Soroush
authors.affiliation: Department of Physiology and Center for Inflammation, Clinical and Translational Lung Research, Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Department of Physiology and Center for Inflammation, Clinical and Translational Lung Research, Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA.
authors.forename: Devon J
authors.initials: DJ
authors.lastname: King
authors.affiliation: Department of Mechanical Engineering, College of Engineering, Temple University, Philadelphia, Pennsylvania, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Department of Mechanical Engineering, College of Engineering, Temple University, Philadelphia, Pennsylvania, USA.
authors.forename: Sudhir
authors.initials: S
authors.lastname: Deosarkar
authors.affiliation: Biomedical Technology, CFD Research Corporation, Huntsville, Alabama, USA; and.
authors.forename: Balabhaskar
authors.initials: B
authors.lastname: Prabhakarpandian
authors.affiliation: Department of Physiology and Center for Inflammation, Clinical and Translational Lung Research, Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA.
authors.forename: Laurie E
authors.initials: LE
authors.lastname: Kilpatrick
authors.affiliation: Department of Mechanical Engineering, College of Engineering, Temple University, Philadelphia, Pennsylvania, USA; mkiani@temple.edu.|Shriners Hospitals Pediatric Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA.
authors.forename: Mohammad F
authors.initials: MF
authors.lastname: Kiani
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000502:physiology; D004195:Disease Models, Animal; D019040:E-Selectin / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000166:cytology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D016209:Interleukin-8 / Q000494:pharmacology; D008168:Lung / Q000276:immunology / Q000382:microbiology; D008297:Male; D046210:Microfluidic Analytical Techniques; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000166:cytology / Q000201:enzymology*; D010446:Peptide Fragments / Q000494:pharmacology; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors / Q000502:physiology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D012212:Rheology; D018805:Sepsis / Q000201:enzymology / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D019040:E-Selectin; D016209:Interleukin-8; D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; D009240:N-Formylmethionine Leucyl-Phenylalanine; D051745:Protein Kinase C-delta
keywords: adhesion; biomimetic; inflammation; leukocytes; transmigration