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使用睡美人转座子增强CD19嵌合抗原受体的表达

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发表于 2016-1-27 21:59:29 | 显示全部楼层 |阅读模式
Manufacture of T cells using the Sleeping Beauty system to enforce expression of a CD19-specific chimeric antigen receptor.
激活T细胞增殖T cells can be reprogrammed to redirect specificity to tumor-associated antigens (TAAs) through the enforced expression of chimeric antigen receptors (CARs). The prototypical CAR is a single-chain molecule that docks with TAA expressed on the cell surface and, in contrast to the T-cell receptor complex, recognizes target cells independent of human leukocyte antigen. The bioprocessing to generate CAR(+) T cells has been reduced to clinical practice based on two common steps that are accomplished in compliance with current good manufacturing practice. These are (1) gene transfer to stably integrate the CAR using viral and nonviral approaches and (2) activating the T cells for proliferation by crosslinking CD3 or antigen-driven numeric expansion using activating and propagating cells (AaPCs). Here, we outline our approach to nonviral gene transfer using the Sleeping Beauty system and the selective propagation of CD19-specific CAR(+) T cells on AaPCs.
使用睡美人转座子增强CD19嵌合抗原受体的表达
T细胞能够被重新编程,通过增强CARs的表达重定向肿瘤相关抗原(TAAs)。原型的CAR是一个单链的分子,与细胞表面表达的TAA相铆钉,与T细胞受体复合物相反,能够识别独立于人类白细胞抗原的靶细胞。生产CAR-T细胞的生物过程已经减少到临床试验为基础的两个步骤,这两个步骤都在当前好的生产操作规范中完成。这些步骤是1)使用病毒或者非病毒的方式进行基因转移,使得CAR能稳定的整合。利用交联抗原CD3或使用活化或增值细胞(AaPCs)来活化T细胞。在这篇文章中,我们利用睡美人转座子系统的非病毒基因转移和在AaPCs中CD19 CAR-T细胞选择性扩增来实现我们的结果:使用睡美人转座子增强CD19嵌合抗原受体的表达
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