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Blood, Vol. 93 No. 6 (March 15), 1999:
pp. 1951-1958
Influence of Monoclonal Antiplatelet Glycoprotein Antibodies on In
Vitro Human Megakaryocyte Colony Formation and Proplatelet Formation
Ryo Takahashi,
Nariko Sekine, and
Toshihiko Nakatake
From the Department of Clinical Hematology, Kyorin University School
of Health Sciences, Tokyo, Japan.
The influence of antiplatelet glycoprotein (GP) antibodies on
megakaryocytopoiesis in patients with idiopathic or immune
thrombocytopenic purpura (ITP) has been well studied. However, the
influence of GP antibodies on proplatelet formation is poorly
understood. Here we investigated whether in vitro human megakaryocyte
colony formation and proplatelet formation are affected by various
monoclonal antiplatelet GP antibodies (MoAb). The megakaryocyte
colony formation inhibition assay was performed by methylcellulose
culture with modifications, using peripheral blood nonadherent
mononuclear cells. The proplatelet formation inhibition assay was
performed by megakaryocytes derived from CD34+ cells,
stimulated with thrombopoietin + stem cell factor, which were then
incubated with antiplatelet GP MoAb for 24 or 48 hours. Anti-GP-Ib
MoAb (CD42b; HIP1) slightly inhibited megakaryocyte colony formation
(P < .05). and strongly inhibited proplatelet formation
(after 24 hours incubation, P < .0002; after 48 hours incubation, P < .0007). Anti-GP-IIb MoAb (CD41; 5B12)
inhibited only proplatelet formation (only after 24 hours incubation,
P < . 03). Anti-integrin v 3
MoAb (CD51/CD61; 23C6) only slightly inhibited colony size
(P < .05). However, anti-GP-IIIa MoAb (CD61; Y2/51) did not
inhibit either colony formation or proplatelet formation. These results
suggest that antiplatelet GP MoAbs have differing effects on in vitro
megakaryocyte colony formation and proplatelet formation.
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