The Expression of LDL-R in CD8 + T Cells Serves as an Early Assessment Parameter for the Production of TCR-T Cells

• Published in: In vivo (Athens) 2023-11, Vol.37 (6), p.2480-2489
• Main Authors: WANG, YI, HAO, YA-NAN, SHEN, MEI-YING, HAN, XIAO-JIAN, WANG, YING-MING, CHEN, SI-YIN, WANG, JUN-FAN, WANG, WANG, LI, TING-TING, JIN, AI-SHUN
• Format: Article
• Language: English
• ISSN: 0258-851X; 1791-7549
• DOI: 10.21873/invivo.13355

BACKGROUND/AIMThe quantity and the phenotypes of desired T cell receptor engineered T (TCR-T) cells in the final cell product determine their in vivo anti-tumor efficacy. Optimization of key steps in the TCR-T cell production process, such as T cell activation, has been shown to improve cell quality.MATERIALS AND METHODSUsing a modified TCR (mTCR) derived from mice transducing PBMCs, we assessed the proportions of low-density lipoprotein receptor (LDL-R) and mTCR expressing cells under the various activation conditions of CD3/CD28-Dynabeads or OKT3 via flow cytometry.RESULTSWe demonstrate that the proportion of T cells expressing LDL-R post activation is positively correlated with the percentage of mTCR+CD8+ T cells with their less differentiated subtypes in the final product. In addition, we show that shifting the CD3/CD28-Dynabeads activation duration from a typical 48 h to 24 h can significantly increase the production of the desired mTCR+CD8+ T cells. Importantly, the percentages of TCR-T cells with less-differentiated phenotypes, namely mTCR central memory T cells (TCM), were found to be preserved with markedly higher efficiency when T cell activation was optimized.CONCLUSIONOur findings suggest that the proportion of LDL-R+ T cells may serve as an early assessment parameter for evaluating TCR-T cell quality, possibly facilitating the functional and economical improvement of current adoptive therapy.