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Differentiation of cellular processes involved in the induction and
maintenance of stimulated neutrophil adherence
D English and TG Gabig
Neutrophil adherence stimulated by phorbol myristate acetate (PMA) was
investigated by quantitating the attachment of 51Cr-labeled neutrophils to
plastic surfaces and to the endothelium of umbilical veins mounted in
compartmentalized Lucite chambers. PMA-induced adherence could be
functionally separated into an induction phase requiring cellular
metabolism and a Mg++ dependent maintenance phase that was independent of
cellular metabolism. Thus, metabolic inhibitors (N-ethylmaleimide, 2-
deoxyglucose) blocked adherence when added to neutrophils prior to PMA, but
did not cause detachment of cells adhering as a consequence of prior
exposure to PMA. PMA failed to induce adherence of neutrophils incubated at
low (0.4 degree C) temperature, but temperature reduction, even for
prolonged periods, did not cause detachment of adherent cells. Thus, the
attractive forces that mediate stimulated adherence persist independently
of any sustained metabolic response to the inducing stimulus. However,
removal of Mg++ from the media above adherent cells resulted in immediate
detachment, indicating that the cation was required for the persistent
expression or maintenance of the attractive forces involved. The extent of
stimulated adherence correlated well with the extent of degranulation when
rates were varied by limiting the incubation time or stimulus
concentration. This correlation was not absolute; in the absence of Mg++,
PMA induced degranulation normally but failed to enhance adherence. To
explain these findings, we investigated the possibility that PMA-stimulated
adherence was maintained by Mg++-dependent cellular adherence molecules
released during exocytosis. Supernatants of stimulated neutrophils were
devoid of adherence-promoting activity, and only weak activity was
recovered in supernatants of mechanically disrupted neutrophils. PMA
effectively stimulated the tight adherence of degranulated neutrophil
cytoplasts to plastic surfaces and did so in the absence of stimulated
granule enzyme release. Thus, conditions have been identified under which
degranulation occurs in the absence of adherence (removal of Mg++) and
adherence occurs without concurrent degranulation. Since neutrophil
cytoplasts do contain some granule products and granule material can be
identified on cytoplast membranes, it is possible that degranulation or
granule products may be involved in the adherent response. However,
hyperadherence was shown to develop in the absence of de novo
degranulation.(ABSTRACT TRUNCATED AT 400 WORDS)
Volume 67,
Issue 5,
pp. 1314-1322,
05/01/1986
Copyright © 1986 by The American Society of Hematology
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