流式细胞术
单克隆抗体
CD14型
吞噬作用
外周血单个核细胞
巨噬细胞
单核细胞
分子生物学
体外
抗体
体内
免疫学
化学
生物
血小板
生物化学
生物技术
作者
Padma K. Narayanan,Nianyu Li
摘要
Abstract Phagocytosis of platelets by monocytes and macrophages is a primary mechanism of platelet clearance in vivo and has been increasingly implicated in playing an important role in thrombocytopenia mediated by monoclonal antibodies intended for therapeutic purposes. In the present article, we describe an in vitro flow cytometry assay to assess the effect of antibody‐mediated platelet phagocytosis by monocytes. Freshly isolated platelets were labeled with a fluorescent probe, 5‐chloromethylfluorescein diacetate (CMFDA) and then co‐cultured with isolated peripheral blood mononuclear cells (PBMCs) from the same donor in the presence of increasing concentrations of a monoclonal antibody drug. After incubation, an increase in CMFDA fluorescence intensity of CD14 positive monocytes was evaluated by flow cytometry as an assessment for drug‐mediated platelet phagocytosis by monocytes. The assay has been evaluated using both human and cynomolgus monkey cells for the prediction of drug‐induced thrombocytopenia. © 2019 by John Wiley & Sons, Inc.
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