Sensitive in vivo imaging of T cells using a membrane-bound Gaussia princeps luciferase.

Sensitive in vivo imaging of T cells using a membrane-bound Gaussia princeps luciferase.
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DOI:
10.1038/nm.1930
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发表时间:
2009-03
期刊:
影响因子:
82.9
通讯作者:
--
中科院分区:
医学1区
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我们开发了一种新的生物发光T细胞成像(BLI)方法,使用一种膜锚定形式的高斯荧光素酶(GLuc),称为extGLuc,我们可以在小鼠和人类的原代T细胞中稳定表达这种酶。在体外,extGLuc+细胞比表达GLuc、Renilla荧光素酶(RLuc)和膜锚定的RLuc(ExtRLuc)的细胞发出更高的生物发光信号。在体内,与GLuc+和RLuc+T细胞相比,小鼠extGLuc+T细胞显示出更高的生物发光信号。将这种成像方法应用于表达肿瘤特异性嵌合抗原受体(CARS)的人T细胞,使我们能够在体内展示CAR介导的T细胞在肿瘤中的积聚,T细胞随时间的持续,以及伴随而来的表达萤火虫荧光素酶(FFLuc)的T细胞和肿瘤细胞的成像。这种灵敏的成像技术已经应用于许多基于活体细胞的研究,在各种各样的小鼠模型上。
We developed a novel approach to bioluminescent T cell imaging (BLI) using a membrane-anchored form of the Gaussia luciferase (GLuc) enzyme, termed extGLuc, which we could stably express in both mouse and human primary T cells. In vitro, extGLuc+ cells emitted significantly higher bioluminescent signal when compared to cells expressing GLuc, Renilla luciferase (RLuc), and membrane-anchored RLuc (extRLuc). In vivo, mouse extGLuc+ T cells exhibited higher bioluminescent signal when compared to GLuc+ and RLuc+ T cells. Application of this imaging approach to human T cells genetically modified to express tumor-specific chimeric antigen receptors (CARs) enabled us to demonstrate in vivo CAR-mediated T cell accumulation in tumor, T cell persistence over time, and concomitant imaging of T cells and tumor cells modified to express firefly luciferase (FFLuc). This sensitive imaging technology has application to many in vivo cell based studies in a wide array of mouse models.
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