Regulation of autoimmune anti-platelet antibody-mediated adhesion of
monocytes to platelet GPIIb/GPIIIa: effect of armed monocytes and the Mac-1
receptor
KB Hymes, MP Schuck and S Karpatkin
New York University Medical School, NY 10016.
Platelet autoantigen-autoantibody-monocyte interaction was studied by
utilization of a specific monoclonal antibody (MoAb) 10E5 to trap and
immobilize the GPIIb-GPIIIa complex on microtiter plates. Peripheral blood
mononuclear cells (PBMC) or purified monocytes formed distinct morphologic
clusters after incubation with immobilized antigen for 18 hours at 37
degrees C. PBMC of 18 and 19 patients with autoimmune thrombocytopenic
purpura (ATP) formed 48 +/- 6.8 (SEM) clusters/well compared with 7.4 +/-
1.0 for control subjects, P less than .001. The number of clusters per well
correlated inversely and exponentially with platelet count, r = -.8, n =
21, indicating that the GPIIb-GPIIIa autoantigen is pathophysiologically
relevant. Binding of ATP PBMC to immobilized GPIIb-GPIIIa could be
inhibited by F(ab')2 fragments of immunoglobulin (Ig) G of ATP patients,
indicating that monocyte IgG bound to autoantigen by its F(ab')2 domain.
Optimal cluster formation could be obtained with normal monocytes if
preincubated with ATP IgG but not with F(ab')2 fragments of ATP IgG,
indicating that ATP IgG binds to monocytes by its Fc domain. Armed
monocytes (ie, normal monocytes preincubated with ATP IgG) bound to
immobilized autoantigen 5.8-fold greater than normal monocytes incubated
with immobilized autoantigen opsonized with ATP IgG. Armed monocyte
adhesion could be inhibited 81% from 18.9 +/- 1.6 to 3.6 +/- 0.5
clusters/well by prior fixation with 0.1% formalin, whereas fixation of IgG
before arming of monocytes was not inhibitory. MoAb MM41, directed against
the alpha m- chain of the Mac-1 adhesive protein receptor of monocytes,
inhibited cluster formation by 79%. Thus, (1) armed monocyte interaction
with autoantigen is considerably more effective than monocyte interaction
with opsonized autoantigen; (2) armed monocyte interaction requires
specific F(ab')2-antigen recognition; and (3) monocyte-autoantigen
interaction requires a secondary nonimmunologic adhesive event.
Volume 75,
Issue 9,
pp. 1813-1819,
05/01/1990
Copyright © 1990 by The American Society of Hematology