Stable transduction of quiescent T cells without induction of cycle progression by a novel lentiviral vector pseudotyped with measles virus glycoproteins
Stable transduction of quiescent T cells without induction of cycle progression by a novel lentiviral vector pseudotyped with measles virus glycoproteins
复制标题
DOI：
10.1182/blood-2008-05-155945
复制
发表时间：
2008-12-15
期刊：
影响因子：
20.3
通讯作者：
Verhoeyen, Els
中科院分区：
文献类型：
--
作者：
Frecha, Cecilia;Costa, Caroline;Verhoeyen, Els
目前慢病毒载体的一个主要局限性是它们不能有效地将基因转移到静止的细胞，如原代T细胞，这阻碍了它们在基因治疗中的应用。在这里，我们产生了表面含有埃德蒙斯顿麻疹病毒(MV)糖蛋白H和F的高滴度LV。它们允许通过MV受体SLAM和CD46进行有效的转导，这两种受体都存在于血液T细胞上。事实上，在IL-7预刺激的T细胞的转导方面，这些H/F显示载体的表现优于远VSV-G-LV。更重要的是，一次接触这些H/F-LV可以在静止的T细胞中进行有效的基因转移，而对于需要细胞周期进入G1b期以进行有效转导的VSV-G-LV来说，这是不允许的。静息记忆(50%)和幼稚T细胞(11%)与H/F-LV的高水平转导似乎主要通过SLAM进行，但并不以细胞周期进入或靶T细胞激活为代价。最后，转导的静息T细胞的幼稚或记忆表型保持不变，细胞因子谱也没有变化，这表明T细胞群体没有倾斜。因此，H/F-LV转导静息T细胞克服了现有慢病毒载体的局限性，可能提高基于T细胞的基因治疗效果。(血。2008年；112：4843-4852)
A major limitation of current lentiviral vectors (LVs) is their inability to govern efficient gene transfer into quiescent cells such as primary T cells, which hampers their application for gene therapy. Here we generated high-titer LVs incorporating Edmonston measles virus (MV) glycoproteins H and F on their surface. They allowed efficient transduction through the MV receptors, SLAM and CD46, both present on blood T cells. Indeed, these H/F-displaying vectors outperformed by far VSV-G-LVs for the transduction of IL-7-prestimulated T cells. More importantly, a single exposure to these H/F-LVs allowed efficient gene transfer in quiescent T cells, which are not permissive for VSV-G-LVs that need cell-cycle entry into the G1b phase for efficient transduction. High-level transduction of resting memory (50%) and naive (11%) T cells with H/F-LVs, which seemed to occur mainly through SLAM, was not at cost of cell-cycle entry or of target T-cell activation. Finally, the naive or memory phenotypes of transduced resting T cells were maintained and no changes in cytokine profiles were detected, suggesting that T-cell populations were not skewed. Thus, H/F-LV transduction of resting T cells overcomes the limitation of current lentiviral vectors and may improve the efficacy of T cell-based gene therapy. (Blood. 2008;112:4843-4852)