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Blood, Vol. 96 No. 1 (July 1), 2000:
pp. 188-194
Morphological analysis of microparticle generation in
heparin-induced thrombocytopenia
Mary Hughes,
Catherine P. M. Hayward,
Theodore E. Warkentin,
Peter Horsewood,
Katherine A. Chorneyko, and
John G. Kelton
From the Departments of Medicine and Pathology, Faculty of Health
Sciences, McMaster University Medical Center, and The
Canadian Blood Services, Hamilton, Ontario, Canada.
Heparin-induced thrombocytopenia (HIT) with thrombosis is a serious
complication of heparin use. HIT sera can generate platelet-derived microparticles, which are produced in a heparin-dependent manner and
are hypothesized to be important initial pathological participants because they promote vascular occlusion. To date, microparticles have
been studied using flow cytometric techniques. However, it is uncertain
whether the small-sized material seen in flow cytometric studies
represents true platelet microparticles shed from activated platelets
or whether they are platelets that have contracted after releasing
their internal components. This report describes a morphological investigation of platelet-derived microparticles in HIT using, among
other techniques, confocal, scanning electron, and transmission electron microscopy. Following incubation with HIT sera, the existence of small membrane-bound vesicles in the milieu of activated platelets was demonstrated. A population of microparticles, expressing
platelet-specific glycoproteins, was separated from platelets by
centrifugation over a sucrose layer. These microparticles had identical
flow cytometric profiles, size heterogeneity, and GPIb
and GPIIb/IIIa staining intensity as the microparticle population in
unfractionated samples. When microparticles were generated in situ and
fixed onto grids, they were demonstrated to be distinct membrane-bound
vesicles that originated near the platelet body and terminal ends of
pseudopods on activated platelets. These microparticles appeared to be
generated by localized swelling, budding, and release. Collectively,
these morphological studies document the existence of true
microparticles in platelets activated by HIT sera. The microparticles
may play an important role in the pathogenesis of HIT.
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