Analysis of human platelet glycoproteins IIb-IIIa and Glanzmann's
thrombasthenia in whole blood by flow cytometry
LK Jennings, RA Ashmun, WC Wang and ME Dockter
Antibodies that bind to human platelet membrane glycoproteins IIb and IIIa
were used to develop methods for analyzing platelet membrane components by
flow cytometry. Platelets were tentatively identified by their
low-intensity light scatter profiles in whole blood or platelet- rich
plasma preparations. Identification of this cell population as platelets
was verified by using platelet-specific antibodies and
fluorescein-conjugated antiimmunoglobulin. Two-parameter analysis of light
scatter versus fluorescence intensity identified greater than 98% of the
cells in the "platelet" light scatter profile as platelets due to their
acquired fluorescence. Both platelet-rich plasma and whole blood were used
to study platelet membrane glycoproteins IIb and IIIa on a single cell
basis in an unwashed system. Prostacycline was included in these
preparations as a precautionary step to inhibit platelet aggregation during
analysis. Flow cytometry is a successful technique for rapid detection of
platelet membrane defects such as Glanzmann's thrombasthenia. Platelets
from Glanzmann's thrombasthenic individuals were readily distinguished from
platelets with normal levels of glycoprotein IIb and IIIa and from
platelets with glycoprotein levels characteristic of heterozygote carriers
of this disorder. This technique provides a sensitive tool for
investigating platelet functional defects due to altered expression or
deficiency of platelet surface proteins.
Volume 68,
Issue 1,
pp. 173-179,
07/01/1986
Copyright © 1986 by The American Society of Hematology
