Lewandrowski Grant K, Magee Ciara N, Mounayar Marwan, Tannous Bakhos A, Azzi Jamil
Experimental Therapeutics and Molecular Imaging Laboratory, Department of Neurology, Neuroscience Center, Massachusetts General Hospital, Boston, MA, USA.
Methods Mol Biol. 2014;1098:211-27. doi: 10.1007/978-1-62703-718-1_17.
Regulatory T cells (Tregs) 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 Treg-based therapy. Consequently, the need to establish the mechanisms that determine Treg 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 Firefly luciferase (Fluc), which permits simultaneous imaging and noninvasive tracking of two different T-cell populations (CD4(+)CD25(+) Tregs and CD4(+)CD25(-) Tcon 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 field of immunology.
调节性T细胞（Tregs）是在包括移植、移植物抗宿主病（GVHD）、癌症和自身免疫等多种免疫介导疾病中研究最为广泛的细胞之一；事实上，基于Treg的疗法的致耐受性潜力备受关注。因此，除了开发监测这些参数的新工具外，确定决定Treg存活和寿命的机制至关重要。利用GVHD小鼠模型和1型糖尿病（T1D）小鼠模型，我们在此描述了一种基于Gluc的双报告系统，并与SEAP和非分泌型萤火虫荧光素酶（Fluc）进行多重组合，通过用携带不同颜色标记的不同慢病毒转导细胞，可在体内同时成像和无创追踪两种不同的T细胞群体（CD4(+)CD25(+) Tregs和CD4(+)CD25(-) Tcon细胞）。这项新技术有望克服目前用于研究体内淋巴细胞存活的传统方法的局限性。此外，这项新技术不仅在自身免疫和同种免疫中有应用，在更广泛的免疫学领域也有应用。
