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Platelet Aggregating Activity Mediated by Thrombin Generation in the NCG Human Neuroblastoma Cell Line¹

Department of Pediatrics, Kyoto Prefectural University of Medicine, Kawaramachi, Kamikyoku, Kyoto, Japan 602

Platelet aggregating activity of the NCG human neuroblastoma cell line was compared with that of the HL-60 human promyelocytic leukemia cell line. NCG, in intact cell suspensions and ultracentrifuged pellets, induced platelet aggregation most significantly in heparinized platelet rich plasma (PRP) containing 2.5 units/ml of heparin, but not in the presence of higher concentrations of heparin or 5 mM ethylenediamine-tetraacetate or in citrated PRP. NCG induced platelet aggregation was also inhibited by hirudin or (2R,4R)-4-methyl-1-[N²-(3-methyl-1,2,3,4-tetrahydro-8-quinolinesulfonyl)-L-arginyl]-2-piperidinecarboxylic acid (MD 805) in the same manner as that of tissue thromboplastin induced platelet aggregation. HL-60 cells did not induce platelet aggregation in our heparinized PRP assay systems; however, after treatment with neuraminidase HL-60 cells became active in aggregating platelets in either heparinized or citrated PRP. NCG demonstrated high procoagulant activity by either intact cell suspensions or ultracentrifuged pellets. The procoagulant activity of NCG was reduced in Factor VII deficient human plasma as it was in the results obtained by tissue thromboplastin. These results suggest that NCG induces platelet aggregation via thrombin generated through procoagulant activity which is shed in association with microvesicles demonstrated in the ultracentrifuged pellets. This type of platelet aggregating activity found in NCG is significantly different from that of HL-60.

¹ This work was supported in part by Grant 59570405 from the Ministry of Education in Japan.

² To whom requests for reprints should be addressed.

Received 5/ 5/86. Revised 9/30/86. Revised 12/31/86. Accepted 1/ 7/87.