Framework for in vivo T cell screens.
Framework for in vivo T cell screens.
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DOI：
10.1084/jem.20230699
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发表时间：
2024-02
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通讯作者：
Lauren E Milling;Samuel C. Markson;Qin Tjokrosurjo;Nicole M Derosia;Ivy S L Streeter;Grant H Hickok;Ashlyn M Lemmen;Thao H Nguyen;Priyamvada Prathima;William Fithian;Marc A. Schwartz;N. Hacohen;John G Doench;M. LaFleur;Arlene H. Sharpe
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作者：
Lauren E Milling;Samuel C. Markson;Qin Tjokrosurjo;Nicole M Derosia;Ivy S L Streeter;Grant H Hickok;Ashlyn M Lemmen;Thao H Nguyen;Priyamvada Prathima;William Fithian;Marc A. Schwartz;N. Hacohen;John G Doench;M. LaFleur;Arlene H. Sharpe
体内T细胞筛选是阐明复杂的免疫机制的有力工具，但在稳健的体内筛选所需的筛选设计参数上缺乏共识：基因文库大小、细胞转移量和小鼠数量。在这里，我们描述了体内T细胞筛选（FITS）的框架，以提供实验和分析指导方针，以确定不同体内环境的最佳参数。作为概念验证，我们使用FITS来优化B16-OVA肿瘤模型中CD 8 + T细胞筛选的参数。我们还在我们的筛选中包括独特的分子标识符（UMI），以（1）提高统计功效和（2）追踪跨多种组织的不同基因敲除（科斯）的T细胞克隆动力学。这些发现为在免疫细胞中进行体内筛选提供了实验和分析框架，并说明了使用UMI进行体内T细胞筛选的案例研究。
In vivo T cell screens are a powerful tool for elucidating complex mechanisms of immunity, yet there is a lack of consensus on the screen design parameters required for robust in vivo screens: gene library size, cell transfer quantity, and number of mice. Here, we describe the Framework for In vivo T cell Screens (FITS) to provide experimental and analytical guidelines to determine optimal parameters for diverse in vivo contexts. As a proof-of-concept, we used FITS to optimize the parameters for a CD8+ T cell screen in the B16-OVA tumor model. We also included unique molecular identifiers (UMIs) in our screens to (1) improve statistical power and (2) track T cell clonal dynamics for distinct gene knockouts (KOs) across multiple tissues. These findings provide an experimental and analytical framework for performing in vivo screens in immune cells and illustrate a case study for in vivo T cell screens with UMIs.