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The role of neutrophil membrane glycoprotein GP-150 in neutrophil adherence
to endothelium in vitro
JM Harlan, PD Killen, FM Senecal, BR Schwartz, EK Yee, RF Taylor, PG Beatty, TH Price and HD Ochs
We have previously described two patients with a congenital defect in
neutrophil function characterized by an inability to form pus. The
patients' neutrophils lack a membrane glycoprotein of mol wt 150,000
daltons (GP-150) on analysis by SDS-PAGE. This glycoprotein is part of a
membrane antigen complex recognized by the murine monoclonal antibody
(MoAb) 60.3. Addition of MoAb 60.3 to normal neutrophils produces defects
in chemotaxis and phagocytosis in vitro similar to those observed in the
patients. Since neutrophil adherence to vascular endothelium is
prerequisite to neutrophil emigration in vivo, we examined the interaction
of the patients' neutrophils and normal neutrophils treated with MoAb 60.3
with cultured endothelium. Adherence was determined as the percentage of
51Cr-labeled purified peripheral blood neutrophils which remained adherent
to plastic wells or endothelial monolayers after a 45-minute incubation at
37 degrees C. The percentage of neutrophils from patient 1 remaining
adherent to uncoated, fibronectin-coated, or laminin-coated plastic was
similar to that observed in normal neutrophils (55% to 84% adherence with
normal neutrophils v 73% to 78% adherence with the patient's neutrophils
and 63% to 82% adherence with MoAb 60.3-treated normal neutrophils). The
adherence of the neutrophils from patient 1 and MoAb 60.3-treated normal
neutrophils to human or bovine endothelium in serum-free medium was also
not significantly different from that observed in normal neutrophils (less
than 10% adherence with normal, MoAb 60.3-treated, and patient
neutrophils). In medium containing 10% autologous or heterologous human
plasma, however, the adherence of neutrophils from patient 1 or MoAb
60.3-treated normal neutrophils to endothelial monolayers was significantly
reduced (35% +/- 7% of normal neutrophils in seven experiments). Although
phorbol myristate acetate (PMA) (10 ng/mL) and calcium ionophore A23187
(10(-5) mol/L) markedly increased the adherence of normal neutrophils to
endothelial monolayers in serum- free medium (40% to 85% adherence),
neither agent increased the adherence of the neutrophils from patient 1 or
normal neutrophils treated with MoAb 60.3 (less than 5% adherence). The
adherence of PMA- activated neutrophils from patient 2 to endothelial
monolayers was also markedly decreased when compared with that of normal
neutrophils. Postsecretory cell-free supernatants from PMA-activated normal
neutrophils failed to augment adherence of neutrophils from patient 1 (less
than 5% adherence).(ABSTRACT TRUNCATED AT 400 WORDS)
Volume 66,
Issue 1,
pp. 167-178,
07/01/1985
Copyright © 1985 by The American Society of Hematology
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