Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
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DOI：
10.1016/j.jim.2011.03.003
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发表时间：
2011-05-31
影响因子：
2.2
通讯作者：
Penichet, Manuel L.
中科院分区：
文献类型：
--
作者：
Helguera, Gustavo;Rodriguez, Jose A.;Penichet, Manuel L.
传统的检测抗体依赖的细胞介导的细胞毒(ADCC)活性的方法依赖于从靶细胞释放可追踪化合物的定量或对人群现象的流式细胞术分析。我们报道了一种直接成像和定量癌细胞ADCC的新方法。采用成像流式细胞术的方法结合了流式细胞术的统计能力和细胞成像的分析优势，为ADCC事件中效应器/靶细胞的相互作用提供了一个新的、更全面的视角。利用这种方法，我们可以定量和详细地显示靶细胞和效应细胞的形态变化、它们的凋亡指数、效应细胞和靶细胞之间的物理相互作用以及胞浆内容物从效应细胞到靶细胞的定向转移。作为一个模型系统，我们使用治疗性抗CD20抗体利妥昔单抗来靶向CFSE标记的Ramos人Burkitt淋巴瘤细胞和CMTPX标记的人单核细胞U-937效应细胞。我们期望，使用不同效应物和靶细胞群体的类似研究可能有助于治疗性抗体的临床前评估，并有助于确定可能对癌症免疫治疗有益的机制。(C)2011爱思唯尔B.V.保留所有权利。
Conventional approaches for the detection of antibody dependent cell-mediated cytotoxicity (ADCC) activity rely on quantification of the release of traceable compounds from target cells or flow cytometry analysis of population-wide phenomena. We report a new method for the direct imaging and quantification of ADCC of cancer cells. The proposed method using imaging flow cytometry combines the statistical power of flow cytometry with the analytical advantages of cell imaging, providing a novel and more comprehensive perspective of effector/target cell interactions during ADCC events. With this method we can quantify and show in detail the morphological changes in target and effector cells, their apoptotic index, the physical interaction between effector and target cells, and a directional transfer of cytosolic contents from effector to target cells. As a model system we used the therapeutic anti-CD20 antibody rituximab to target CFSE labeled Ramos human Burkitt's lymphoma cells, to CMTPX-labeled human monocytic U-937 effector cells. We expect that similar studies using different effector and target cell populations may contribute to the pre-clinical evaluation of therapeutic antibodies and help to identify mechanisms that could be beneficial in the immunotherapy of cancer. (C) 2011 Elsevier B.V. All rights reserved.