Individual Motile CD4(+) T Cells Can Participate in Efficient Multikilling through Conjugation to Multiple Tumor Cells.
Individual Motile CD4(+) T Cells Can Participate in Efficient Multikilling through Conjugation to Multiple Tumor Cells.
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DOI：
10.1158/2326-6066.cir-14-0195
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发表时间：
2015-05
影响因子：
10.1
通讯作者：
Varadarajan N
中科院分区：
文献类型：
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作者：
Liadi I;Singh H;Romain G;Rey-Villamizar N;Merouane A;Adolacion JR;Kebriaei P;Huls H;Qiu P;Roysam B;Cooper LJ;Varadarajan N
用于B细胞恶性肿瘤研究治疗的转基因T细胞表达CD19特异性嵌合抗原受体(CAR)，构成了一个不同的群体，它们持续并参与连续杀伤肿瘤细胞的能力是治疗成功的预测因素。我们在Nanowell网格中实施了TimeLapse成像显微镜(TIMING)，以提供直接证据，表明CD4+CAR+T细胞(CAR4细胞)可以通过同时与多个肿瘤细胞结合而参与多重杀伤。对CAR4细胞和CD8+CAR+T细胞(CAR8细胞)的比较表明，虽然CAR4细胞可以参与杀伤和多重杀伤，但它们的速度较慢，可能是因为颗粒酶B含量较低。值得注意的是，在这两组T细胞中，以其高运动性识别的一小部分单独的T细胞亚群显示了对单个肿瘤细胞的有效杀伤。通过对多个杀伤细胞和单个杀伤细胞CAR+T细胞的比较，发现T细胞的凋亡倾向和动力学似乎受到功能结合数量的调节。分离的T细胞在与单个肿瘤细胞结合时，发生了快速的、更高频率的凋亡，这种作用在CAR8细胞上更为明显。我们的结果提示，CAR+T细胞参与多重杀伤的能力应根据其抵抗激活诱导的细胞死亡(AICD)的能力来评估。我们预计，这一时机可能被用来快速确定T细胞群体的效力，并可能促进下一代CAR+T细胞的设计和制造，提高疗效。
T cells genetically modified to express a CD19-specific chimeric antigen receptor (CAR) for the investigational treatment of B-cell malignancies comprise a heterogeneous population, and their ability to persist and participate in serial killing of tumor cells is a predictor of therapeutic success. We implemented Timelapse Imaging Microscopy In Nanowell Grids (TIMING) to provide direct evidence that CD4+CAR+ T cells (CAR4 cells) can engage in multi-killing via simultaneous conjugation to multiple tumor cells. Comparisons of the CAR4 cells and CD8+CAR+ T cells (CAR8 cells) demonstrate that while CAR4 cells can participate in killing and multi-killing, they do so at slower rates, likely due to the lower Granzyme B content. Significantly, in both sets of T cells, a minor sub-population of individual T cells identified by their high motility, demonstrated efficient killing of single tumor cells. By comparing both the multi-killer and single killer CAR+ T cells it appears that the propensity and kinetics of T-cell apoptosis was modulated by the number of functional conjugations. T cells underwent rapid apoptosis, and at higher frequencies, when conjugated to single tumor cells in isolation and this effect was more pronounced on CAR8 cells. Our results suggest that the ability of CAR+ T cells to participate in multi-killing should be evaluated in the context of their ability to resist activation induced cell death (AICD). We anticipate that TIMING may be utilized to rapidly determine the potency of T-cell populations and may facilitate the design and manufacture of next-generation CAR+ T cells with improved efficacy.