Berkowitz R, Ilves H, Lin W Y, Eckert K, Coward A, Tamaki S, Veres G, Plavec I
Systemix Inc., Palo Alto, California 94304, USA.
J Virol. 2001 Apr;75(7):3371-82. doi: 10.1128/JVI.75.7.3371-3382.2001.
Because lentiviruses are able to infect nondividing cells, these viruses might be utilized in gene therapy applications where the target cell does not divide. However, it has been suggested that the introduction of primate lentivirus sequences, particularly those of human immunodeficiency virus, into human cells may pose a health risk for the patient. To avoid this concern, we have constructed gene transfer systems based on a nonprimate lentivirus, bovine immunodeficiency virus. A panel of vectors and packaging constructs was generated and analyzed in a transient expression system for virion production and maturation, vector expression and encapsidation, and envelope protein pseudotyping. Virion preparations were also analyzed for transduction efficiency in a panel of human and nonhuman primary cells and immortalized cell lines. The virion preparations transduced most of the target cell types, with efficiencies up to 90% and with titers of unconcentrated virus up to 5 x 10(5) infectious doses/ml. In addition, infection of nondividing human cells, including unstimulated hematopoietic stem cells and irradiated endothelial cells, was observed.
由于慢病毒能够感染非分裂细胞，这些病毒可用于靶细胞不分裂的基因治疗应用中。然而，有人提出将灵长类慢病毒序列，特别是人类免疫缺陷病毒的序列引入人类细胞可能会给患者带来健康风险。为避免这一问题，我们构建了基于非灵长类慢病毒——牛免疫缺陷病毒的基因转移系统。生成了一组载体和包装构建体，并在瞬时表达系统中分析了病毒粒子的产生和成熟、载体表达和包装以及包膜蛋白假型化情况。还分析了病毒粒子制剂在一组人类和非人类原代细胞以及永生化细胞系中的转导效率。病毒粒子制剂转导了大多数靶细胞类型，效率高达90%，未浓缩病毒的滴度高达5×10⁵ 感染剂量/毫升。此外，还观察到了对非分裂人类细胞的感染，包括未刺激的造血干细胞和经辐照的内皮细胞。
