https://scholars.lib.ntu.edu.tw/handle/123456789/564221
標題: | Analysis of human platelet glycoprotein IIb-IIIa by fluorescein isothiocyanate-conjugated disintegrins flow cytometry | 作者: | Liu C.-Z. Wang Y.-W. Shen M.-C. TUR-FU HUANG |
公開日期: | 1994 | 卷: | 72 | 期: | 6 | 起(迄)頁: | 919-925 | 來源出版物: | Thrombosis and Haemostasis | 摘要: | Disintegrins are a group of snake venom peptides which inhibit human platelet aggregation by acting as glycoprotein IIb-IIIa (GPIIb-IIIa) antagonists. They are cysteine-rich, Arg-Gly-Asp(RGD)-containing peptides, and bind to GPIIb-IIIa complex on platelet membrane with a very high affinity (Kd, 10-7 ~ 10-8 M). In this study, we analyzed GPIIb-IIIa complex on platelet membrane by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated disintegrins as probes. Of these FITC-conjugated disintegrins, FITC-Rhodostomin is the most sensitive probe because Rhodostomin was conjugated with more FITC molecules than Trigramin and Halysin were. The binding fluorescence intensity of FITC-Trigramin (FITC-Tg), FITC-Halysin (FITC-Hy) and FITC-Rhodostomin (FITC-Rn) was measured in both resting and ADP-activated platelets of diluted human platelet-rich plasma. The binding fluorescence of FITC-disintegrins was abolished by EDTA and 7E3, a monoclonal antibody against GPIIb-IIIa. ADP markedly increased the fluorescence intensity of FITC-Tg and FITC-Hy bound on platelets especially when lower doses of these probes were used, whereas it had little effect on that of FITC-Rn. Therefore, FITC-Tg and FITC-Hy can be used for the detection of the activated platelets as noted by a higher ratio of fluorescence intensity (approx. 2-4) between ADP-activated and resting platelets as compared with that (approx. 1-1.3) in the case of FITC-Rn as the probe. The platelets from three patients with Glanzmann's thrombasthenia were probed with FITC-disintegrins. As a result these three patients could be classified as type I thrombasthenia based on the extremely low level of GPIIb-IIIa detected by this method ( < 5% of normal value). |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/564221 | ISSN: | 3406245 | DOI: | 10.1055/s-0038-1648984 | SDG/關鍵字: | adenosine diphosphate; edetic acid; fibrinogen receptor; fluorescein isothiocyanate; monoclonal antibody; peptide; snake venom; trigramin; article; binding affinity; case report; controlled study; disease classification; flow cytometry; fluorescence; Glanzmann disease; human; human cell; priority journal; protein binding; thrombocyte activation; thrombocyte aggregation; thrombocyte membrane |
顯示於: | 藥理學科所 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。