Beta 2-integrin and L-selectin are obligatory receptors in neutrophil aggregation.
We have recently found that antibodies to L-selectin, the homing receptor on neutrophils, are as effective as those to beta 2-integrin at blocking formyl peptide-stimulated aggregation. Therefore, we investigated the requirements for expression of L-selectin and beta 2-integrin on adjacent cells during aggregation. Fluorescence flow cytometry allowed characterization of aggregates on the basis of size and color, as well as antibody binding to these two adhesive molecules. Formyl peptide-stimulated aggregate formation was measured for individual populations fluorescently labeled red (LDS-751) or green (CD44-FITC), and interpopulation red-green cell conjugates. Blocking either the beta 2-integrin or L-selectin adhesive epitope with monoclonal antibody on individual cell populations resulted in an approximately 50% reduction in two-color aggregation as compared with that in unblocked samples. Shedding the L-selectin on a cell population by preincubation with complexes of lipopolysaccharide and its plasma membrane binding protein also decreased aggregation to a control population by approximately 50%. We examined the aggregation of neutrophils from patients genetically deficient in beta 2-integrin and clinically leukocyte adhesion deficient (LAD). LAD adhesion to normal neutrophils was dependent on the expression of L-selectin on LAD cells and beta 2-integrin on normal cells. Thus, the minimum requirement for adhesion between two mixed populations of neutrophils was that one population expressed the beta 2-integrin and the other expressed the L-selectin adhesive epitope.
Simon
S I
SI
Department of Pathology, University of New Mexico Cancer Center, School of Medicine, Albuquerque.
Rochon
Y P
YP
Lynam
E B
EB
Smith
C W
CW
Anderson
D C
DC
Sklar
L A
LA
D000818:Animals; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000502:physiology*; D002449:Cell Aggregation; D002918:Chymotrypsin / Q000494:pharmacology; D006801:Humans; D016023:Integrins / Q000032:analysis / Q000502:physiology*; D007700:Kinetics; D019041:L-Selectin; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D009504:Neutrophils / Q000502:physiology*
D015815:Cell Adhesion Molecules; D016023:Integrins; D008070:Lipopolysaccharides; D019041:L-Selectin; D002918:Chymotrypsin