Rolling on E- or P-selectin induces the extended but not high-affinity conformation of LFA-1 in neutrophils.
Human blood neutrophils rolling on E- or P-selectin reduced their rolling velocity when intercellular adhesion molecule (ICAM)-1 was available. Similar to mouse neutrophils, this was dependent on P-selectin glycoprotein ligand 1 (PSGL1), alpha(L)beta(2) integrin, the Src family tyrosine kinase FGR and spleen tyrosine kinase SYK. Blocking phospholipase C or p38 MAP kinase attenuated, but did not abolish the velocity reduction. To test expression of integrin activation epitopes, we adapted an immobilized reporter assay and developed a new homogeneous microfluidics-based reporter antibody binding assay. Rolling on E- or P-selectin induced the extension reporter epitopes KIM127 and NKI-L16, but not the high affinity reporter epitope monoclonal antibody (mAb) 24. This enabled rolling neutrophils to bind to immobilized extension reporter, but not activation reporter antibodies and allowed binding of soluble KIM127 during rolling. We conclude that human neutrophil rolling on E- or P-selectin induces the extended alpha(L)beta(2) integrin conformation through signaling triggered by PSGL-1 engagement.
Kuwano
Yoshihiro
Y
Division of Inflammation Biology, La Jolla Institute for Allergy & Immunology, CA, USA.
Spelten
Oliver
O
Zhang
Hong
H
Ley
Klaus
K
Zarbock
Alexander
A
D019040:E-Selectin / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism / Q000502:physiology; D036904:Leukocyte Rolling / Q000276:immunology / Q000502:physiology*; D016169:Lymphocyte Function-Associated Antigen-1 / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000502:physiology; D009504:Neutrophils / Q000378:metabolism* / Q000502:physiology; D019007:P-Selectin / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding / Q000502:physiology; D011487:Protein Conformation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism / Q000502:physiology; D013379:Substrate Specificity; D000072377:Syk Kinase; D010738:Type C Phospholipases / Q000378:metabolism / Q000502:physiology; D020298:U937 Cells; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism / Q000502:physiology; D019061:src-Family Kinases / Q000378:metabolism / Q000502:physiology
D019040:E-Selectin; D047908:Intracellular Signaling Peptides and Proteins; D016169:Lymphocyte Function-Associated Antigen-1; D008562:Membrane Glycoproteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C000606897:Syk protein, mouse; D019061:src-Family Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D010738:Type C Phospholipases
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