A simplified method for the rapid fluorometric assessment of antibody-dependent cell-mediated cytotoxicity
A simplified method for the rapid fluorometric assessment of antibody-dependent cell-mediated cytotoxicity
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DOI：
10.1016/j.jim.2005.09.018
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发表时间：
2006-01-20
影响因子：
2.2
通讯作者：
Robert-Guroff, M
中科院分区：
文献类型：
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作者：
Gómez-Román, VR;Florese, RH;Robert-Guroff, M
我们证明，致命的细胞溶解试验可以适应为快速和荧光抗体依赖性细胞毒性试验（RFADCC）。RFADCC依赖于在添加抗体和效应细胞之前，用膜染料（PKH-26）和活力染料（CFSE）对靶细胞进行双重染色。我们使用RFADCC来评估单克隆抗体2gi2和人血清介导的剂量依赖性和包膜特异性抗人类免疫缺陷病毒（HIV） ADCC反应。利用该方法，我们还检测了感染致病性SIVmac251的恒河猴的早期抗猴免疫缺陷病毒（SIV） ADCC反应。重要的是，RFADCC在监测用复制腺病毒为基础的艾滋病候选疫苗免疫黑猩猩和恒河猴所引起的抗hiv和抗siv ADCC反应方面进一步有用。与标准的铬释放测定相比，RFADCC提供了更高的细胞杀伤读数，并且在允许使用冷冻和新鲜效应细胞方面具有优势，从而促进了测定的标准化。因此，RFADCC是一种简单、可靠和高度敏感的方法，可用于评估单克隆抗体的ADCC活性，以及由HIV或SIV感染或艾滋病候选疫苗引起的ADCC反应。(c) 2005 Elsevier B.V.版权所有
We demonstrate that the FATAL cytolysis assay can be adapted into a rapid and fluorometric antibody-dependent cellular cytotoxicity assay (RFADCC). The RFADCC relies on double-staining target cells with a membrane dye (PKH-26) and a viability dye (CFSE) prior to the addition of antibody and effector cells. We used the RFADCC to assess dose-dependent and envelope-specific anti-human immunodeficiency virus (HIV) ADCC responses mediated by monoclonal antibody-2GI2 and human sera. Using the assay, we also detected early anti-simian immunodeficiency virus (SIV) ADCC responses in rhesus macaques infected with pathogenic SIVmac251. Importantly, the RFADCC was further useful in monitoring anti-HIV and anti-SIV ADCC responses elicited by immunizing chimpanzees and rhesus macaques with replicating adenovirus-based AIDS vaccine candidates. In comparison to the standard chromium release assay, the RFADCC provides a higher cell killing readout and is advantageous in allowing use of viably frozen as well as fresh effector cells, thus facilitating assay standardization. The RFADCC is therefore a simple, reliable, and highly sensitive method that can be applied to assess the ADCC activity of monoclonal antibodies as well as ADCC responses elicited by HIV or SIV infection or by AIDS vaccine candidates. (c) 2005 Elsevier B.V. All rights reserved.