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Activation mechanisms of adherent human neutrophils
I Ginis and AI Tauber
William B. Castle Hematology Research Laboratory, Boston City Hospital, MA.
The mechanism by which unstimulated human neutrophils initiate a
respiratory burst on adherence to a surface has been examined. When
neutrophils adhere to a plastic surface, they immediately generate a
sustained burst of superoxide (O2-). However, this respiratory burst is not
initiated by adherence alone, since neutrophils attached to fibronectin
fail to mount a response. Adhesion to plastic is calcium (Ca2+)
independent, but O2- production requires Ca2(+)-containing buffer in the
initiation phase, that is, during adhesion and the early phase of O2-
production. The Ca2(+)-dependent step was shown to involve protein kinase C
(PK-C) in that the O2- production, but not adherence, was blocked with
1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), and PK-C was found to
translocate from the cytosol to the membrane on adhesion. Furthermore, it
may be inferred that this translocation results in the generation of a Ca2+
independent form of PK-C, PK-M, since leupeptin, which inhibits the
generation of PK-M, also blocked O2- production. This finding was
corroborated by showing that after 5 minutes in a Ca2(+)-containing buffer,
enough time to initiate O2- production and PK-C translocation, Ca2+ is no
longer required for sustained O2- release. These results, in aggregate,
demonstrate that neutrophils are activated by adhesion to plastic to
generate O2-, a PK- C-dependent process that appears to involve a
Ca2(+)-independent form of the kinase, PK-M. Why adherent neutrophils
generate a respiratory burst on plastic and not fibronectin surfaces
probably reflects activation of distinct receptors, whose nature must still
be defined. Another issue to address is the priming effect of adhesion,
since cells adherent to plastic- or fibronectin-coated surfaces have an
enhanced O2- response to formylmethionyl-leucine-phenylalanine (FMLP)
compared with neutrophils stimulated in suspension. This may relate to
increased Ca2+ mobilization, an important mediator of priming for FMLP
responses. Thus, adhesion as a priming event does not necessarily initiate
cell effector function, and the further elucidation of the plastic and
fibronectin models suggests a means of characterizing the crucial event
that control neutrophil activation.
Volume 76,
Issue 6,
pp. 1233-1239,
09/15/1990
Copyright © 1990 by The American Society of Hematology
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