Gibbon ape leukemia virus (GALV) can infect a wide variety of cells but fails to infect most cells derived from laboratory mice. Transduction of human hematopoietic stem cells with GALV retroviral vectors is more efficient than with amphotropic vectors. In this study, a Moloney murine leukemia virus-gibbon ape leukemia virus (MoMLV-GALV) vector was constructed by replacing the natural env gene of the full-length Moloney MLV genome with the GALV env gene. To monitor viral transmission by green fluorescent protein (GFP) expression, internal ribosomal entry site-enhanced GFP (IRES-EGFP) was positioned between the GALV env gene and the 3' untranslated region (3' UTR) to obtain pMoMLV-GALV-EGFP. The MoMLV-GALV-EGFP vector was able to replicate with high titer in TE671 human rhabdomyosarcoma cells and U-87 human glioma cells. To evaluate the potential of the MoMLV-GALV vector as a therapeutic agent, the gene for the fusogenic envelope G glycoprotein of vesicular stomatitis virus (VSV-G) was incorporated into the vector. Infection with the resulting MoMLV-GALV-VSV-G vector resulted in lysis of the U-87 cells due to syncytium formation. Syncytium formation was also observed in the transfected human prostate cancer cell line LNCaP after extended cultivation of cells. In addition, we deleted the GALV env gene from the MoMLV-GALV-VSV-G vector to improve viral genome stability. This MoMLV-VSV-G vector is also replication competent and induces syncytium formation in 293T, HT1080, TE671 and U-87 cells. These results suggest that replication of the MoMLV-GALV-VSV-G vector or MoMLV-VSV-G vector may directly lead to cytotoxicity. Therefore, the vectors developed in this study are potentially useful tools for cancer gene therapy.

中文翻译：

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具有复制能力的逆转录病毒载体的构建，用于表达用于癌症基因治疗的VSV-G包膜糖蛋白。

长臂猿白血病病毒（GALV）可以感染多种细胞，但不能感染大多数源自实验室小鼠的细胞。用GALV逆转录病毒载体转导人造血干细胞比使用两性载体更有效。在这项研究中，莫洛尼氏鼠白血病病毒-长臂猿白血病病毒（MoMLV-GALV）载体是通过用GALV env基因替换全长莫洛尼氏MLV基因组的天然env基因而构建的。为了监测绿色荧光蛋白（GFP）表达的病毒传播，将内部核糖体进入位点增强的GFP（IRES-EGFP）置于GALV env基因和3'非翻译区（3'UTR）之间，以获得pMoMLV-GALV-EGFP 。MoMLV-GALV-EGFP载体能够在TE671人横纹肌肉瘤细胞和U-87人神经胶质瘤细胞中高滴度复制。为了评估MoMLV-GALV载体作为治疗剂的潜力，将水泡性口腔炎病毒的融合膜G糖蛋白（VSV-G）基因整合到该载体中。由于合胞体形成，感染所得的MoMLV-GALV-VSV-G载体导致U-87细胞裂解。长时间培养细胞后，在转染的人前列腺癌细胞系LNCaP中也观察到合胞体形成。此外，我们从MoMLV-GALV-VSV-G载体中删除了GALV env基因，以提高病毒基因组的稳定性。此MoMLV-VSV-G载体也具有复制能力，并在293T，HT1080，TE671和U-87细胞中诱导合胞体形成。这些结果表明，MoMLV-GALV-VSV-G载体或MoMLV-VSV-G载体的复制可能直接导致细胞毒性。因此，