Shuttle of lentiviral vectors via transplanted cells in vivo
Shuttle of lentiviral vectors via transplanted cells in vivo
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DOI：
10.1038/sj.gt.3302384
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发表时间：
2005-01-01
期刊：
影响因子：
5.1
通讯作者：
Martin, U
中科院分区：
文献类型：
--
作者：
Blömer, U;Gruh, I;Martin, U
慢病毒载体已被证明是一种有效的方法，在体外和体内稳定的基因转移。其应用领域不仅包括体内移植后的细胞标记和追踪，而且还包括用于基因治疗的生物活性蛋白的稳定递送。然而，各种细胞需要在制备后立即移植，例如，以防止细胞死亡、分化或去分化。尽管这些细胞通常在慢病毒转导后洗涤数次，但可能存在病毒载体穿梭通过移植细胞的风险，导致受体细胞的不期望的体内转导。我们研究了感染性慢病毒颗粒是否通过离体慢病毒转导细胞传播。为此，我们探索了潜在的病毒穿梭通过离体慢病毒转导的心肌细胞在体外和移植到大脑和外周肌肉。我们证明，即使经过大量洗涤，感染性病毒载体颗粒可以在细胞悬液中检测到。这些慢病毒载体颗粒能够在transwell实验中粘附靶细胞。此外，传递的载体颗粒在体内稳定地转导受体中枢神经系统和肌肉的驻留细胞。我们的结果慢病毒载体穿梭通过转导的心肌细胞是显着的离体基因治疗和慢病毒细胞示踪，特别是在移植模型和共培养系统中的干细胞分化的调查。
Lentiviral vectors have turned out to be an efficient method for stable gene transfer in vitro and in vivo. Not only do fields of application include cell marking and tracing following transplantation in vivo, but also the stable delivery of biological active proteins for gene therapy. A variety of cells, however, need immediate transplantation after preparation, for example, to prevent cell death, differentiation or dedifferentiation. Although these cells are usually washed several times following lentiviral transduction, there may be the risk of viral vector shuttle via transplanted cells resulting in undesired in vivo transduction of recipient cells. We investigated whether infectious lentiviral particles are transmitted via ex vivo lentivirally transduced cells. To this end, we explored potential viral shuttle via ex vivo lentivirally transduced cardiomyocytes in vitro and following transplantation into the brain and peripheral muscle. We demonstrate that, even after extensive washing, infectious viral vector particles can be detected in cell suspensions. Those lentiviral vector particles were able to transduce target cells in transwell experiments. Moreover, transmitted vector particles stably transduced resident cells of the recipient central nervous system and muscle in vivo. Our results of lentiviral vector shuttle via transduced cardiomyocytes are significant for both ex vivo gene therapy and for lentiviral cell tracing, in particular for investigation of stem cell differentiation in transplantation models and co-cultivation systems.