New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population
New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population
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DOI：
10.1182/blood-2003-06-2070
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发表时间：
2004-04-01
期刊：
影响因子：
20.3
通讯作者：
Falkenburg, JHF
中科院分区：
文献类型：
--
作者：
Jedema, I;van der Werff, NM;Falkenburg, JHF
为了临床评价细胞免疫治疗的疗效，有必要分析T细胞对白血病原发靶细胞群的效应功能，这些原发靶细胞群通常是由不同成熟阶段的白血病细胞引起的。一种合适的检测方法不仅应该能够定量分析常规的Cr-51释放法所测量的快速细胞死亡诱导，而且还应该能够在几天内而不是几小时内对T细胞诱导的凋亡的较慢执行途径进行定量分析，而不是用这种方法无法测量的数小时。此外，它应该剖析不同靶细胞亚群对T细胞诱导的细胞死亡的不同易感性，并表征能够产生恶性后代的恶性前体细胞。为了满足这些要求，我们开发了一种新的检测方法，该方法基于目标细胞群体的羧基荧光素二乙酸酯琥珀酰亚胺酯(CFSE)标记，结合特定细胞群体的抗体染色和添加荧光微球，以定量监测在更长时间内发生的目标细胞死亡，最长可达5天。这一新的检测方法有助于分析不同靶细胞群体中不同细胞类型的区别识别，并允许同时评估存活的靶细胞对相关细胞因子的反应的增殖状态。(C)2004年，由美国血液病学会提供。
For the clinical evaluation of the efficacy of cellular immunotherapy it is necessary to analyze the effector functions of T cells against primary leukemic target cell populations which are usually considerably heterogeneous caused by differential maturation stages of the leukemic cells. An appropriate assay should not only allow the quantitative analysis of rapid cell death induction as measured by the conventional Cr-51 release assay but also of the more slowly executing pathways of T-cell-induced apoptosis occurring within days Instead of hours which cannot be measured using this method. Furthermore, It should dissect the differential susceptibility to T-cell-induced cell death of various target cell subpopulations and characterize the malignant precursor cells capable of producing malignant progeny. To fulfill these requirements we developed a new assay based on carboxyfluorescein diacetate succinimidyl ester (CFSE) labeling of the target cell population combined with antibody staining of specific cell populations and addition of fluorescent microbeads to quantitatively monitor target cell death occurring within a longer time frame up to at least 5 days. This new assay facilitates the analysis of differential recognition of distinct cell types within a heterogeneous target cell population and allows simultaneously evaluation of the proliferative status of surviving target cells in response to relevant cytokines. (C) 2004 by The American Society of Hematology.