Department of Bioengineering, University of Washington, Seattle, WA, USA.
Ben Towne Center for Childhood Cancer Research, Seattle Children's Research Institute, Seattle, WA, USA.
Nat Biomed Eng. 2019 Oct;3(10):783-795. doi: 10.1038/s41551-019-0411-6. Epub 2019 Jun 17.
Chimeric antigen receptor T-cell therapies using defined product compositions require high-purity T-cell isolation systems that, unlike immunomagnetic positive enrichment, are inexpensive and leave no trace on the final cell product. Here, we show that DNA aptamers (generated with a modified cell-SELEX procedure to display low-nanomolar affinity for the T-cell marker CD8) enable the traceless isolation of pure CD8 T cells at low cost and high yield. Captured CD8 T cells are released label-free by complementary oligonucleotides that undergo toehold-mediated strand displacement with the aptamer. We also show that chimeric antigen receptor T cells manufactured from these cells are comparable to antibody-isolated chimeric antigen receptor T cells in proliferation, phenotype, effector function and antitumour activity in a mouse model of B-cell lymphoma. By employing multiple aptamers and the corresponding complementary oligonucleotides, aptamer-mediated cell selection could enable the fully synthetic, sequential and traceless isolation of desired lymphocyte subsets from a single system.
使用定义明确的产品成分的嵌合抗原受体 T 细胞疗法需要高纯度的 T 细胞分离系统，与免疫磁阳性富集不同，这些系统既便宜又不会在最终细胞产品上留下痕迹。在这里，我们展示了 DNA 适体（通过改良的细胞 SELEX 程序生成，对 T 细胞标志物 CD8 的亲和力低至纳摩尔级）能够以低成本和高产量无痕迹地分离纯 CD8 T 细胞。通过与适体进行 toehold 介导的链置换的互补寡核苷酸，可无标记释放捕获的 CD8 T 细胞。我们还表明，从这些细胞制造的嵌合抗原受体 T 细胞在增殖、表型、效应功能和抗肿瘤活性方面与抗体分离的嵌合抗原受体 T 细胞相当在 B 细胞淋巴瘤的小鼠模型中。通过使用多个适体和相应的互补寡核苷酸，适体介导的细胞选择可以从单个系统中完全合成、顺序和无痕迹地分离所需的淋巴细胞亚群。
