title: Neutrophils exposed to A. phagocytophilum under shear stress fail to fully activate, polarize, and transmigrate across inflamed endothelium.
abstract: Anaplasma phagocytophilum is an obligate intracellular bacterium that has evolved mechanisms to hijack polymorphonuclear neutrophil (PMN) receptors and signaling pathways to bind, infect, and multiply within the host cell. E-selectin is upregulated during inflammation and is a requisite endothelial receptor that supports PMN capture, rolling, and activation of integrin-mediated arrest. Ligands expressed by PMN that mediate binding to endothelium via E-selectin include sialyl Lewis x (sLe(x))-expressing ligands such as P-selectin glycoprotein ligand-1 (PSGL-1) and other glycolipids and glycoproteins. As A. phagocytophilum is capable of binding to sLe(x)-expressing ligands expressed on PMN, we hypothesized that acute bacterial adhesion to PMN would subsequently attenuate PMN recruitment during inflammation. We assessed the dynamics of PMN recruitment and migration under shear flow in the presence of a wild-type strain of A. phagocytophilum and compared it with a strain of bacteria that binds to PMN independent of PSGL-1. Acute bacterial engagement with PMN resulted in transient PMN arrest and minimal PMN polarization. Although the wild-type pathogen also signaled activation of beta2 integrins and elicited a mild intracellular calcium flux, downstream signals including PMN transmigration and phosphorylation of p38 mitogen-activated protein kinase (MAPK) were inhibited. The mutant strain bound less well to PMN and failed to activate beta2 integrins and induce a calcium flux but did result in decreased PMN arrest and polarization that may have been partially mediated by a suppression of p38 MAPK activation. This model suggests that A. phagocytophilum binding to PMN under shear flow during recruitment to inflamed endothelium interferes with normal tethering via E-selectin and navigational signaling of transendothelial migration.
authors.affiliation: Department of Biomedical Engineering, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
authors.forename: U Y
authors.initials: UY
authors.lastname: Schaff
authors.forename: K A
authors.initials: KA
authors.lastname: Trott
authors.forename: S
authors.initials: S
authors.lastname: Chase
authors.forename: K
authors.initials: K
authors.lastname: Tam
authors.forename: J L
authors.initials: JL
authors.lastname: Johns
authors.forename: J A
authors.initials: JA
authors.lastname: Carlyon
authors.forename: D C
authors.initials: DC
authors.lastname: Genetos
authors.forename: N J
authors.initials: NJ
authors.lastname: Walker
authors.forename: S I
authors.initials: SI
authors.lastname: Simon
authors.forename: D L
authors.initials: DL
authors.lastname: Borjesson
mesh_terms: D041081:Anaplasma phagocytophilum / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D000818:Animals; D001422:Bacterial Adhesion; D018821:CD18 Antigens / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D020013:Calcium Signaling; D016764:Cell Polarity; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000276:immunology* / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D007700:Kinetics; D007739:L Cells; D036904:Leukocyte Rolling; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D009154:Mutation; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D010766:Phosphorylation; D013314:Stress, Mechanical; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D018821:CD18 Antigens; D015820:Cadherins; D018836:Inflammation Mediators; D053583:Interleukin-1beta; D008562:Membrane Glycoproteins; C084882:P-selectin ligand protein; D014409:Tumor Necrosis Factor-alpha; D048051:p38 Mitogen-Activated Protein Kinases
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