III Medical Clinic at the University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany.
TRON (Translational Oncology at the University Medical Center), Johannes Gutenberg University Mainz, Mainz, Germany.
Gene Ther. 2014 May;21(5):533-8. doi: 10.1038/gt.2014.25. Epub 2014 Apr 3.
Adoptive T-cell transfer for cancer immunotherapy requires genetic modification of T cells with recombinant T-cell receptors (TCRs). Amphotropic retroviral vectors (RVs) used for TCR transduction for this purpose are considered safe in principle. Despite this, TCR-coding and packaging vectors could theoretically recombine to produce replication competent vectors (RCVs), and transduced T-cell preparations must be proven free of RCV. To eliminate the need for RCV testing, we transduced human T cells with ecotropic RVs so potential RCV would be non-infectious for human cells. We show that transfection of synthetic messenger RNA encoding murine cationic amino-acid transporter 1 (mCAT-1), the receptor for murine retroviruses, enables efficient transient ecotropic transduction of human T cells. mCAT-1-dependent transduction was more efficient than amphotropic transduction performed in parallel, and preferentially targeted naive T cells. Moreover, we demonstrate that ecotropic TCR transduction results in antigen-specific restimulation of primary human T cells. Thus, ecotropic RVs represent a versatile, safe and potent tool to prepare T cells for the adoptive transfer.
过继性 T 细胞转移的癌症免疫疗法需要对 T 细胞进行基因修饰，使其带有重组 T 细胞受体 (TCR)。为此目的而使用的双嗜性逆转录病毒载体 (RV) 在原则上被认为是安全的。尽管如此，TCR 编码和包装载体理论上可以重组产生复制能力的载体 (RCV)，并且转导的 T 细胞制剂必须证明不含 RCV。为了消除对 RCV 检测的需求，我们用嗜性 RV 转导人 T 细胞，使潜在的 RCV 对人细胞无感染性。我们表明，编码鼠阳离子氨基酸转运蛋白 1(mCAT-1)的合成信使 RNA 的转染使人类 T 细胞能够有效地进行瞬时嗜性转导。mCAT-1 依赖性转导比同时进行的双嗜性转导更有效，并且优先靶向幼稚 T 细胞。此外，我们证明了嗜性 TCR 转导导致原发性人 T 细胞的抗原特异性再刺激。因此，嗜性 RV 是一种通用、安全且有效的工具，可用于制备用于过继转移的 T 细胞。
