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Continuous activation and deactivation of integrin CD11b/CD18 during de
novo expression enables rolling neutrophils to immobilize on platelets
S Sheikh and GB Nash
Department of Physiology, The Medical School, The University of Birmingham,
Birmingham, UK.
In an in vitro flow model, unstimulated neutrophils rolled steadily over a
surface coated with platelets, until superfusion of the chemotactic peptide
formyl-methionyl-leucyl-phenylalanine (fMLP) caused a dose-dependent
(10(-11) to 10(-7) mol/L) transition from rolling to stationary attachment
in seconds, followed more slowly by neutrophil shape change and spreading
on the surface, However, at low concentrations of Ca2+ and Mg2+ (0.1 mmol/L
and 0.05 mmol/L, respectively, rather than physiologic 1 mmol/L and 0.5
mmol/L), neutrophils first halted but then started to roll again and to
detach from the surface over 5 to 10 minutes. At the low cation
concentration, stopping was largely inhibited by antibodies to the
neutrophil integrins CD18 or CD11b, but not CD11a. When neutrophils were
pretreated with antibodies to CD11b or CD18 in 1 mmol/L Ca2+ 0.5 mmol/L
Mg2+, stopping was not prevented but delayed. However, if antibodies were
also included with the superfused fMLP, stopping was inhibited, and
detachment followed. This indicates that CD11b/CD18 was newly expressed
during shape change and mediated the second phase of neutrophil
immobilization and spreading in a cation-dependent manner. Prestimulated
neutrophils also bound to platelets and spread, but immobilization was
blocked if they were perfused with antibody to CD18 or CD11b or with low
Ca2+ and Mg2+. Examining the cation-dependence further, it was evident that
the presence of Mg2+ was essential for integrin-mediated adhesion and that
the Mg2+ concentration determined whether immobilization could be
maintained or was transient. Continuous superfusion of fMLP was also
essential for maintenance of stable adhesion and spreading. Thus,
activation of constitutive CD11b/CD18 rapidly and reversibly converted
rolling to stationary attachment, whereas maintenance of adhesion and
neutrophil spreading required continual expression of additional CD11b/CD18
that was only functional at physiologic Mg2+. Continual activation and
deactivation of CD11b/CD18 during de novo expression could mediate
immobilization and onward migration of neutrophils in vivo, and activated
platelets appear capable of supporting this process as well as endothelial
cells.
Volume 87,
Issue 12,
pp. 5040-5050,
06/15/1996
Copyright © 1996 by The American Society of Hematology
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