Signaling functions of L-selectin in neutrophils: alterations in the cytoskeleton and colocalization with CD18.
Ligation and clustering of L-selectin by Ab ("cross-linking") or physiologic ligands results in activation of diverse responses that favor enhanced microvascular sequestration and emigration of neutrophils. The earliest responses include a rise in intracellular calcium, enhanced tyrosine phosphorylation, and activation of extracellular signal-regulated kinases. Additionally, cross-linking of L-selectin induces sustained shape change and activation of beta2 integrins, leading to neutrophil arrest under conditions of shear flow. In this report, we examined several possible mechanisms whereby transmembrane signals from L-selectin might contribute to an increase in the microvascular retention of neutrophils and enhanced efficiency of emigration. In human peripheral blood neutrophils, cross-linking of L-selectin induced alterations in cellular biophysical properties, including a decrease in cell deformability associated with F-actin assembly and redistribution, as well as enhanced adhesion of microspheres bound to beta2 integrins. L-selectin and the beta2 integrin became spatially colocalized as determined by confocal immunofluorescence microscopy and fluorescence resonance energy transfer. We conclude that intracellular signals from L-selectin may enhance the microvascular sequestration of neutrophils at sites of inflammation through a combination of cytoskeletal alterations leading to cell stiffening and an increase in adhesiveness mediated through alterations in beta2 integrins.
Simon
S I
SI
Department of Pediatrics, Section of Leukocyte Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Cherapanov
V
V
Nadra
I
I
Waddell
T K
TK
Seo
S M
SM
Wang
Q
Q
Doerschuk
C M
CM
Downey
G P
GP
D000199:Actins / Q000378:metabolism; D000276:Adjuvants, Immunologic / Q000502:physiology; D000328:Adult; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001696:Biomechanical Phenomena; D018821:CD18 Antigens / Q000378:metabolism* / Q000502:physiology; D002448:Cell Adhesion / Q000276:immunology; D002465:Cell Movement / Q000276:immunology; D003599:Cytoskeleton / Q000276:immunology* / Q000378:metabolism; D004735:Energy Transfer; D005374:Filtration; D005434:Flow Cytometry; D006801:Humans; D007106:Immune Sera / Q000378:metabolism; D018189:Immunomagnetic Separation; D019041:L-Selectin / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D015398:Signal Transduction / Q000276:immunology*; D013050:Spectrometry, Fluorescence
D000199:Actins; D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D018821:CD18 Antigens; D007106:Immune Sera; D019041:L-Selectin; D009240:N-Formylmethionine Leucyl-Phenylalanine