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Contact-induced neutrophil activation by platelets in human cell
suspensions and whole blood
A Ruf, RF Schlenk, A Maras, E Morgenstern and H Patscheke
Institute for Clinical Chemistry, Klinikum Mannheim, University of
Heidelberg, Germany.
Platelet-dependent activation of polymorphonuclear neutrophils (PMNL) was
investigated with a lumi-aggregometer in heparinized whole blood and
platelet-PMNL suspensions. The lumi-aggregometer allowed us to
simultaneously monitor increases in impedance or light transmission as
consequences of platelet aggregation and luminol-enhanced chemiluminescence
(CL) as a measure of the oxidative burst in PMNL. Aggregation and
platelet-PMNL contacts were also checked by light and electron microscopy.
In whole blood, adenosine diphosphate (ADP) and the thromboxane A2 mimetic
U 46619 induced the aggregation (increase in impedance) and the CL, which
were both suppressed by EDTA, arginyl- glycyl-aspartyl-serine (RGDS)
peptide, and the absence of stirring. In contrast, FMLP caused only CL that
was unaffected by EDTA, RGDS peptide, and nonstirring. Similar observations
were obtained with mixed suspensions containing washed platelets and PMNL
at their physiologic concentrations. ADP, U 46619, and thrombin induced
both aggregation (increase in light transmission) and CL, whereas FMLP
caused CL but only very weak aggregation. Exogenous fibrinogen strongly
enhanced the effects of ADP and U 46619. Iloprost, EDTA, RGDS peptide, red
blood cell (RBC) ghosts, and nonstirring inhibited the effects induced by
the platelet agonists, but were ineffective on the CL induced by FMLP.
Treatment of platelets with aspirin did not affect the CL of PMNL induced
by platelets. Microscopic examination, the requirements of stirring, Ca2+,
and fibrinogen, and the inhibitory effects of RGDS peptide and RBC ghosts
show that stimulated platelets activate PMNL in a contact-dependent manner
that depends on fibrinogen binding. This was confirmed by the
immunochemical demonstration of fibrinogen (but not of fibronectin) in the
contact spaces between activated platelets and PMNL. Because supernatants
and lysates of resting or thrombin- stimulated platelets did not induce the
CL of PMNL, soluble agonists did not appear to be involved. Nonstimulated
washed platelets also caused CL of PMNL that required stirring and Ca2+ and
was inhibited by RBC ghosts. No CL occurred in unstimulated stirred whole
blood, suggesting that a preactivation of platelets during the preparation
may be responsible for the effects of unstimulated washed platelets. The
results show that platelets provide a strong stimulus for PMNL that
requires intercellular contact. Fibrinogen exposure on the platelet surface
seems to be necessary for the activation of PMNL by stimulated platelets.
Volume 80,
Issue 5,
pp. 1238-1246,
09/01/1992
Copyright © 1992 by The American Society of Hematology
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