A Novel Cell-based Luciferase Reporter Platform for the Development and Characterization of T-Cell Redirecting Therapies and Vaccine Development

被引:3
作者
Grailer, Jamison [1 ]
Cheng, Zhijie Jey [1 ]
Hartnett, Jim [1 ]
Slater, Michael [1 ]
Fan, Frank [1 ]
Cong, Mei [1 ]
机构
[1] Promega Corp, Dept Res, 2800 Woods Hollow Rd, Madison, WI 53711 USA
关键词
T cell; TCR-T; vaccine; cell-based bioassay; reporter bioassay; CLASS-II MOLECULE; ANTIGEN; RECEPTOR; COMPLEX; CANCER; PRODUCTS; PEPTIDE; TCRS; CD4;
D O I
10.1097/CJI.0000000000000453
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
T-cell immunotherapies are promising strategies to generate T-cell responses towards tumor-derived or pathogen-derived antigens. Adoptive transfer of T cells genetically modified to express antigen receptor transgenes has shown promise for the treatment of cancer. However, the development of T-cell redirecting therapies relies on the use of primary immune cells and is hampered by the lack of easy-to-use model systems and sensitive readouts to facilitate candidate screening and development. Particularly, testing T-cell receptor (TCR)-specific responses in primary T cells and immortalized T cells is confounded by the presence of endogenous TCR expression which results in mixed alpha/beta TCR pairings and compresses assay readouts. Herein, we describe the development of a novel cell-based TCR knockout (TCR-KO) reporter assay platform for the development and characterization of T-cell redirecting therapies. CRISPR/Cas9 was used to knockout the endogenous TCR chains in Jurkat cells stably expressing a human interleukin-2 promoter-driven luciferase reporter gene to measure TCR signaling. Reintroduction of a transgenic TCR into the TCR-KO reporter cells results in robust antigen-specific reporter activation compared with parental reporter cells. The further development of CD4/CD8 double-positive and double-negative versions enabled low-avidity and high-avidity TCR screening with or without major histocompatibility complex bias. Furthermore, stable TCR-expressing reporter cells generated from TCR-KO reporter cells exhibit sufficient sensitivity to probe in vitro T-cell immunogenicity of protein and nucleic acid-based vaccines. Therefore, our data demonstrated that TCR-KO reporter cells can be a useful tool for the discovery, characterization, and deployment of T-cell immunotherapy.
引用
收藏
页码:96 / 106
页数:11
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