Beta2-integrins mediate stable adhesion in collisional interactions between neutrophils and ICAM-1-expressing cells.
The aggregation of human neutrophils in suspension has features that are analogous to their attachment to activated endothelium in that both involve selectin and beta2-integrin adhesion receptors. For the collisional interaction that forms neutrophil aggregates in suspension, there is a tethering step in which L-selectin on neutrophils binds PSGL-1. At relatively low shear rates (100-200 s(-1)) firm adhesion is mediated in equal measure by LFA-1 binding to ICAM-3, and Mac-1 binding to an as yet undefined ligand. In this report we used a mouse melanoma cell line expressing an estimated 700,000 ICAM-1 (CD54) to examine the relative roles of LFA-1 and Mac-1 over the kinetics of heterotypic cell adhesion in shear mixed suspensions. Neither heterotypic nor homotypic neutrophil aggregates formed with application of shear alone. However, the rate of aggregation peaked within seconds of chemotactic stimulation. In contrast to homotypic aggregation, neither L-selectin nor its O-glycoprotein ligands on neutrophils contributed to heterotypic adhesion. Adhesion was inhibited in a dose-dependent manner as ICAM-1 was titrated with blocking mAb. A direct interaction between LFA-1 and ICAM-1 was preferred over the first minute of stimulation, whereas at later times adhesion was supported equally by Mac-1. Activation with MnCl2 also favored participation of the constitutively expressed LFA-1. Application of defined shear in a cone and plate viscometer showed that adhesion to the ICAM-1 cells decreased from a maximum level to baseline as shear rate increased up to 400 s(-1) in a manner typical of integrin adhesion alone. In contrast, homotypic aggregation supported by the transition from selectin to integrin binding exhibited an increase in efficiency up to 800 s(-1). The pathophysiological significance of receptor site density and duration of contact in collisional interactions relevant to leukocyte recruitment compared to leukocyte-endothelial cell interactions on surfaces is discussed.
Lynam
E
E
Cytometry, Cancer Research and Treatment Center, University of New Mexico Health Science Center, Albuquerque 87131, USA.
Sklar
L A
LA
Taylor
A D
AD
Neelamegham
S
S
Edwards
B S
BS
Smith
C W
CW
Simon
S I
SI
D000818:Animals; D018821:CD18 Antigens / Q000502:physiology*; D002448:Cell Adhesion; D002449:Cell Aggregation / Q000502:physiology; D002450:Cell Communication; D002712:Chlorides / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000502:physiology*; D019041:L-Selectin / Q000502:physiology; D016169:Lymphocyte Function-Associated Antigen-1 / Q000502:physiology; D016177:Macrophage-1 Antigen / Q000502:physiology; D017895:Manganese Compounds / Q000494:pharmacology; D008546:Melanoma, Experimental / Q000473:pathology; D008562:Membrane Glycoproteins / Q000502:physiology; D051379:Mice; D009504:Neutrophils / Q000166:cytology* / Q000378:metabolism; D013314:Stress, Mechanical; D014407:Tumor Cells, Cultured
D018821:CD18 Antigens; D002712:Chlorides; D016169:Lymphocyte Function-Associated Antigen-1; D016177:Macrophage-1 Antigen; D017895:Manganese Compounds; D008562:Membrane Glycoproteins; C084882:P-selectin ligand protein; D018799:Intercellular Adhesion Molecule-1; D019041:L-Selectin; C025340:manganese chloride