Simultaneous In Vivo Monitoring of Regulatory and Effector T Lymphocytes Using Secreted Gaussia Luciferase, Firefly Luciferase, and Secreted Alkaline Phosphatase
Simultaneous In Vivo Monitoring of Regulatory and Effector T Lymphocytes Using Secreted Gaussia Luciferase, Firefly Luciferase, and Secreted Alkaline Phosphatase
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DOI：
10.1007/978-1-62703-718-1_17
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发表时间：
2014-01-01
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影响因子：
--
通讯作者：
Azzi, Jamil
中科院分区：
文献类型：
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作者：
Lewandrowski, Grant K.;Magee, Ciara N.;Azzi, Jamil
调节性T细胞（Treg）是在多种免疫介导的病症（包括移植和移植物抗宿主病（GVHD）、癌症和自身免疫）中研究最广泛的细胞之一;实际上，对基于Treg的疗法的致耐受性潜力存在极大兴趣。因此，除了开发新的工具来监测这些参数之外，建立确定T reg存活和寿命的机制的需要是至关重要的。使用GVHD的小鼠模型和1型糖尿病（T1 D）的小鼠模型，我们在本文中描述了基于Gluc并与SEAP和非分泌型萤火虫荧光素酶（Fluc）多路复用的双报告子系统，它允许同时成像和非侵入性跟踪两种不同的T细胞群通过用具有不同颜色特征的不同慢病毒转导细胞，在体内将细胞（CD 4 + CD 25 + T-con和CD 4（+）CD 25-T-con细胞）转染。这项新技术有望克服目前可用于研究体内淋巴细胞存活的常规方法的局限性。此外，这种新技术不仅在自身免疫和同种免疫中有应用，而且在更广泛的免疫学领域也有应用。
Regulatory T cells (T regs) are amongst the most widely studied cells in a variety of immune-mediated conditions, including transplantation and Graft Versus Host Disease (GVHD), cancer and autoimmunity; indeed, there is great interest in the tolerogenic potential of T reg -based therapy. Consequently, the need to establish the mechanisms that determine T reg survival and longevity, in addition to developing new tools to monitor these parameters, is paramount. Using both a mouse model of GVHD and a mouse model of Type 1 Diabetes (T1D), we describe herein a dual reporter system based on Gluc and multiplexed with SEAP and non-secreted Firefl y luciferase (Fluc), which permits simultaneous imaging and noninvasive tracking of two different T-cell populations (CD4 + CD25 + T-regs and CD4(+)CD25-T-con cells) in vivo by transducing the cells with different lentiviruses bearing distinct color signatures. This new technology promises to overcome the limitations of the conventional methods currently available to study lymphocyte survival in vivo. Furthermore, this novel technique has applications not only in autoimmunity and alloimmunity, but also in the wider fi eld of immunology.