Cell Therapy Group, Cancer Research UK Department of Medical Oncology, University of Manchester, Manchester Academic Health Science Centre, Manchester, UK.
J Gene Med. 2010 Feb;12(2):129-36. doi: 10.1002/jgm.1421.
HIV-1 fails to successfully infect mouse T cells as a result of several blocks in the viral replication cycle. We investigated whether this also impacted on the use of HIV-1 derived lentiviral vectors for stable gene transfer into mouse T cells.
Freshly isolated primary mouse T cells were immediately mixed with lentiviral vectors encoding an enhanced green fluorescent protein marker gene and transduction frequency was determined after 5 days of culture.
Optimal transduction required both mouse T cell activation and cytokine support. Furthermore, transduction was also dependent upon the promoter chosen, with the rank order of potency being PGK > EF1 > SFFV > CMV. HIV-1 lentiviral vectors also efficiently transduced cytokine-stimulated T cells (in the absence of antibody driven T cell activation), albeit with a lower level of transgene expression compared to fully-activated T cells.
The present study demonstrates that primary mouse T cells can be efficiently transduced with HIV-1 lentiviral vectors, opening up prospects for their use in mouse models of gene-modified adoptive cellular therapy.
由于 HIV-1 复制周期中的几个环节受阻，其无法成功感染小鼠 T 细胞。我们研究了这是否也会影响 HIV-1 衍生的慢病毒载体在稳定基因转移到小鼠 T 细胞中的应用。
新鲜分离的原代小鼠 T 细胞立即与编码增强型绿色荧光蛋白标记基因的慢病毒载体混合，并在培养 5 天后测定转导频率。
最佳转导需要小鼠 T 细胞的激活和细胞因子的支持。此外，转导还取决于所选择的启动子，其效力的顺序为 PGK > EF1 > SFFV > CMV。HIV-1 慢病毒载体也能有效地转导细胞因子刺激的 T 细胞（无需抗体驱动的 T 细胞激活），但与完全激活的 T 细胞相比，转导基因的表达水平较低。
本研究表明，原代小鼠 T 细胞可以有效地被 HIV-1 慢病毒载体转导，为其在基因修饰过继细胞治疗的小鼠模型中的应用开辟了前景。
