The success of chimeric antigen receptor (CAR) T cell therapies for treating leukemia has resulted in a booming interest for the technology. Expression of a CAR in T cells allows redirection of their natural cytolytic activity toward cells presenting a specific designated surface antigen. Although CAR T cell therapies have thus far shown promising results mostly in B cell malignancy trials, interest in their potential to treat other diseases is on the rise, including using CAR T cells to control human immunodeficiency virus infection. The assessment of CAR T cell potency toward specific targets in vitro is a critical preclinical step. In this study, we describe novel assays that monitor the cytotoxicity of candidate CAR T cells toward simian immunodeficiency virus (SIV) infected CD4 T cells. The assays involve live cell imaging using a fluorescence microscopy system that records in real time the disappearance or appearance of targets infected with SIV carrying a fluorescent protein gene. The assays are highly reproducible, and their rapid turn around and reduced cost present a significant advance regarding the efficient preclinical evaluation of CAR T cell constructs and are broadly applicable to potential human diseases that could benefit from CAR T cell therapy.

中文翻译：

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评估新型抗猿免疫缺陷病毒嵌合抗原受体 T 细胞效力的实时杀伤分析

嵌合抗原受体 (CAR) T 细胞疗法治疗白血病的成功引起了人们对该技术的浓厚兴趣。CAR 在 T 细胞中的表达允许将它们的天然溶细胞活性重新定向到呈递特定指定表面抗原的细胞。尽管迄今为止 CAR T 细胞疗法主要在 B 细胞恶性肿瘤试验中显示出有希望的结果，但人们对其治疗其他疾病的潜力的兴趣正在上升，包括使用 CAR T 细胞来控制人类免疫缺陷病毒感染。体外评估 CAR T 细胞对特定靶标的效力是关键的临床前步骤。在这项研究中，我们描述了监测候选 CAR T 细胞对猴免疫缺陷病毒 (SIV) 感染的 CD4 T 细胞的细胞毒性的新检测方法。该分析涉及使用荧光显微镜系统进行活细胞成像，该系统实时记录被携带荧光蛋白基因的 SIV 感染的靶标的消失或出现。这些测定具有高度可重复性，其快速周转和降低成本在 CAR T 细胞构建体的有效临床前评估方面取得了重大进展，并且广泛适用于可能受益于 CAR T 细胞治疗的潜在人类疾病。