A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells.
A Nontoxic Transduction Enhancer Enables Highly Efficient Lentiviral Transduction of Primary Murine T Cells and Hematopoietic Stem Cells.
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DOI：
10.1016/j.omtm.2018.08.002
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发表时间：
2018-09-21
期刊：
影响因子：
--
通讯作者：
Six E
中科院分区：
文献类型：
--
作者：
Delville M;Soheili T;Bellier F;Durand A;Denis A;Lagresle-Peyrou C;Cavazzana M;Andre-Schmutz I;Six E
慢病毒载体已成为基于造血干细胞（HSC）或T细胞的基因治疗的有效、安全的治疗工具。然而，在临床前模型中监测转导的细胞仍然具有挑战性，因为与γ逆转录病毒载体相比，用慢病毒载体转导鼠原代T细胞的效率低。当慢病毒载体将用于临床试验时，随后在临床前概念验证中使用这一点不被认为是相关的。因此，迫切需要开发一种用于小鼠细胞的具有慢病毒载体的有效转导方案。在这里，我们描述了一个优化的协议，其中无毒的转导增强剂（Lentiboost），使有效的转导原代小鼠T细胞与慢病毒载体。优化的方案结合了低毒性和高转导效率。我们用VSV-G假型慢病毒载体实现了小鼠CD 4+和CD 8 + T细胞的高水平转导，转导的T细胞的表型没有变化，其在培养中稳定且寿命长。该增强子还增加了鼠HSC的转导。因此，使用这种新的转导增强剂克服了慢病毒载体在临床前实验中的局限性，并应促进基于慢病毒载体的策略从实验室到临床的翻译。
Lentiviral vectors have emerged as an efficient, safe therapeutic tool for gene therapy based on hematopoietic stem cells (HSCs) or T cells. However, the monitoring of transduced cells in preclinical models remains challenging because of the inefficient transduction of murine primary T cells with lentiviral vectors, in contrast to gammaretroviral vectors. The use of this later in preclinical proof of concept is not considered as relevant when a lentiviral vector will be used in a clinical trial. Hence, there is an urgent need to develop an efficient transduction protocol for murine cells with lentiviral vectors. Here, we describe an optimized protocol in which a nontoxic transduction enhancer (Lentiboost) enables the efficient transduction of primary murine T cells with lentiviral vectors. The optimized protocol combines low toxicity and high transduction efficiency. We achieved a high-level transduction of murine CD4+ and CD8+ T cells with a VSV-G-pseudotyped lentiviral vector with no changes in the phenotypes of transduced T cells, which were stable and long-lived in culture. This enhancer also increased the transduction of murine HSCs. Hence, use of this new transduction enhancer overcomes the limitations of lentiviral vectors in preclinical experiments and should facilitate the translation of strategies based on lentiviral vectors from the bench to the clinic.
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