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Platelet/polymorphonuclear leukocyte interaction in dynamic conditions:
evidence of adhesion cascade and cross talk between P-selectin and the beta
2 integrin CD11b/CD18
V Evangelista, S Manarini, S Rotondo, N Martelli, R Polischuk, JL McGregor, G de Gaetano and C Cerletti
G. Bizzozero Laboratory of Platelet and Leukocyte Pharmacology, Consorzio
Mario Negri Sud, Santa Maria Imbaro, Italy.
Adhesion between platelets and polymorphonuclear leukocytes (PMN) is a key
event in thrombosis and inflammation. Double color fluorescence- activated
cell sorter (FACS) analysis was used to determine the extent and kinetics
of adhesion of thrombin-activated platelets to resting or activated PMN
when mixed cell populations were incubated in dynamic conditions. Activated
platelets bound very rapidly to PMN. Mixed cell conjugates reached a
maximum at 1 minute and were reversible within 10 minutes. Platelet/PMN
adhesion required both Ca2+ and Mg2+ and was markedly increased by the
presence of Mn2+. The latter made mixed cell conjugates stable up to 10
minutes. Adhesion of platelets required metabolic activity of PMN and was
abolished by tyrosine kinase inhibitors. Furthermore, adhesion of platelets
to PMN resulted in binding of a monoclonal antibody (MoAb 24) known as beta
2 integrins "activation reporter." When PMN were activated by exogenous
stimuli, the adhesion of platelets was markedly increased: fMLP induced a
rapid and transient effect, while PMA resulted in a slower, but stable,
increase in mixed conjugates formation. The hypothesis that activated PMN
beta 2 integrins are able to bind a counter-receptor on platelets was
directly demonstrated by the increase of mixed cell conjugates following
PMN treatment with KIM127 and KIM185, two anti-CD18 antibodies able to
induce the active conformation of beta 2 integrins. Consistently, two other
anti-CD18, as well as an anti-CD11b inhibitory antibody abolished
platelet/PMN adhesion. PMN beta 2 integrin activation was not the only
mechanism for activated platelet/PMN adhesion to occur: indeed, this
phenomenon could also be inhibited by two anti-P-selectin antibodies.
Resting platelets did not adhere to resting PMN, but markedly adhered to
fMLP- or PMA-activated PMN. Resting platelet/fMLP-activated PMN adhesion
was abolished by anti-CD18 antibodies, but not by anti-P-selectin
antibodies. In conclusion, activated platelet/PMN interaction can be
modeled as an adhesion cascade involving a P-selectin-dependent recognition
step and a functional signal. The latter proceeds through tyrosine kinase
activation and enables a beta 2 integrin-dependent adhesion to a not yet
identified counter-receptor constitutively expressed on platelet surface.
Volume 88,
Issue 11,
pp. 4183-4194,
12/01/1996
Copyright © 1996 by The American Society of Hematology
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