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Human neutrophils lose their surface Fc gamma RIII and acquire Annexin V
binding sites during apoptosis in vitro
CH Homburg, M de Haas, AE von dem Borne, AJ Verhoeven, CP Reutelingsperger and D Roos
Central Laboratory of the Netherlands Red Cross Blood Transfusion Service,
Amsterdam.
We have previously reported that neutrophilic granulocytes rapidly release
part of their Fc gamma RIII from the plasma membrane upon in vitro
activation, probably by proteolytic cleavage. In plasma and other body
fluids, released or soluble Fc gamma RIII has been found in considerable
amounts. In the present study, neutrophils were kept in maintenance culture
for 18 to 24 hours. Forty percent of the neutrophils completely lost Fc
gamma RIII, and the remainder of the cells showed a 60% decrease in Fc
gamma RIII expression on their surface. Released Fc gamma RIII was detected
in the culture supernatant. Nevertheless, more than 90% of the cells was
viable as judged by hydrolysis of fluorescein diacetate. The presence of
interferon gamma, granulocyte colony-stimulating factor, or granulocyte-
macrophage colony-stimulating factor, but not interleukin-3 (IL-3), IL- 6,
or IL-8, in the culture medium increased the number of cells that still
expressed Fc gamma RIII. We found that this loss of Fc gamma RIII was not
the result of cell activation but correlated strongly with apoptosis. The
Fc gamma RIII-negative subpopulation exhibited typical morphologic changes,
such as nuclear condensation and DNA fragmentation. Furthermore, this
subpopulation appeared to have acquired the property of binding Annexin V,
a calcium-dependent, phospholipid-binding protein with high affinity for
phosphatidylserine. The external exposure of this phospholipid by cells has
been reported to occur during apoptosis. The property of Annexin V binding
was not shared by the nonapoptotic, Fc gamma RIII-positive subpopulation.
In this respect, we identified binding of Annexin V as an convenient marker
for apoptotic cells. Our results indicate that soluble Fc gamma RIII in
body fluids might be derived for a large part from neutrophils undergoing
apoptosis in the tissues.
Volume 85,
Issue 2,
pp. 532-540,
01/15/1995
Copyright © 1995 by The American Society of Hematology
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