In vitro immunotherapy potency assays using real-time cell analysis.
In vitro immunotherapy potency assays using real-time cell analysis.
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DOI：
10.1371/journal.pone.0193498
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Xi B
中科院分区：
文献类型：
--
作者：
Cerignoli F;Abassi YA;Lamarche BJ;Guenther G;Santa Ana D;Guimet D;Zhang W;Zhang J;Xi B
随着人们对免疫效应细胞和靶肿瘤细胞之间分子相互作用的认识不断加深，再加上完善的基因治疗方法，正在产生新的癌症免疫疗法，在临床上对固体和液体肿瘤都有显著的疗效。虽然免疫疗法在治疗某些癌症方面具有巨大的希望，但在临床转化为许多其他类型的癌症以及最小化不良反应方面仍然存在重大挑战。因此，迫切需要在体外和体内进行功能效价测定，以模拟免疫细胞与肿瘤细胞的复杂相互作用，并可用于快速测试不同免疫治疗方法的疗效，无论是小分子、生物制剂、细胞疗法还是它们的联合疗法。在此，我们报告了xCELLigence实时细胞溶解体外效价测定的发展，该检测使用细胞阻抗连续监测靶肿瘤细胞在接受不同类型治疗时的活力。包含集成金微电极的专用微滴板能够在包括效应细胞、抗体、小分子等在内的异质混合物中选择性地监测粘附目标肿瘤细胞的数量、大小和表面附着强度。通过表面捆绑的方法，还可以监测液体癌的杀伤情况。以NK92效应细胞为例，RTCA效价测定的结果与基于图像的测定和流式细胞术的终点数据非常相关。几种效应细胞，如PBMC、NK、CAR-T，以及生物分子，如靶向肿瘤细胞上表达的EpCAM蛋白的Bi-specific T cell Engagers （BiTEs）和针对免疫检查点抑制剂PD-1的阻断抗体，都被测试和验证。使用专门设计的xCELLigence免疫治疗软件，计算KT50（杀死50%靶肿瘤细胞所需时间）和细胞溶解%等定量参数，并用于比较不同试剂的相对疗效。总之，我们的结果表明，xCELLigence平台非常适合效价分析，提供了高重复性的定量评估，大大简化了工作流程。
A growing understanding of the molecular interactions between immune effector cells and target tumor cells, coupled with refined gene therapy approaches, are giving rise to novel cancer immunotherapeutics with remarkable efficacy in the clinic against both solid and liquid tumors. While immunotherapy holds tremendous promise for treatment of certain cancers, significant challenges remain in the clinical translation to many other types of cancers and also in minimizing adverse effects. Therefore, there is an urgent need for functional potency assays, in vitro and in vivo, that could model the complex interaction of immune cells with tumor cells and can be used to rapidly test the efficacy of different immunotherapy approaches, whether it is small molecule, biologics, cell therapies or combinations thereof. Herein we report the development of an xCELLigence real-time cytolytic in vitro potency assay that uses cellular impedance to continuously monitor the viability of target tumor cells while they are being subjected to different types of treatments. Specialized microtiter plates containing integrated gold microelectrodes enable the number, size, and surface attachment strength of adherent target tumor cells to be selectively monitored within a heterogeneous mixture that includes effector cells, antibodies, small molecules, etc. Through surface-tethering approach, the killing of liquid cancers can also be monitored. Using NK92 effector cells as example, results from RTCA potency assay are very well correlated with end point data from image-based assays as well as flow cytometry. Several effector cells, i.e., PBMC, NK, CAR-T were tested and validated as well as biological molecules such as Bi-specific T cell Engagers (BiTEs) targeting the EpCAM protein expressed on tumor cells and blocking antibodies against the immune checkpoint inhibitor PD-1. Using the specifically designed xCELLigence immunotherapy software, quantitative parameters such as KT50 (the amount of time it takes to kill 50% of the target tumor cells) and % cytolysis are calculated and used for comparing the relative efficacy of different reagents. In summary, our results demonstrate the xCELLigence platform to be well suited for potency assays, providing quantitative assessment with high reproducibility and a greatly simplified work flow.
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影响因子：
10.1
作者：
Harshman LC;Drake CG;Choueiri TK
通讯作者：
Choueiri TK
DOI：
10.1016/j.omto.2016.11.003
发表时间：
2017-03-17
期刊：
Molecular therapy oncolytics
影响因子：
--
作者：
Dyer A;Di Y;Calderon H;Illingworth S;Kueberuwa G;Tedcastle A;Jakeman P;Chia SL;Brown A;Silva MA;Barlow D;Beadle J;Hermiston T;Ferguson DJ;Champion B;Fisher KD;Seymour LW
通讯作者：
Seymour LW
影响因子：
1.8
作者：
Glamann, Joakim;Hansen, Anker Jon
通讯作者：
Hansen, Anker Jon
影响因子：
12.4
作者：
Dubey P
通讯作者：
Dubey P
DOI：
10.3390/bios5020199
发表时间：
2015-04-16
期刊：
Biosensors
影响因子：
--
作者：
Kho D;MacDonald C;Johnson R;Unsworth CP;O'Carroll SJ;du Mez E;Angel CE;Graham ES
通讯作者：
Graham ES