Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.
PLoS One. 2012;7(8):e42455. doi: 10.1371/journal.pone.0042455. Epub 2012 Aug 16.
Recombinant interferon-γ (IFNγ) production in cultured lentivirus (LV) was explored for inhibition of target virus in cells co-infected with adenovirus type 5 (Ad5). The ability of three different promoters of CMV, EF1α and Ubiquitin initiating the enhanced green fluorescence protein (GFP) activities within lentiviruses was systematically assessed in various cell lines, which showed that certain cell lines selected the most favorable promoter driving a high level of transgenic expression. Recombinant IFNγ lentivirus carrying CMV promoter (LV-CMV-IFNγ) was generated to co-infect 293A cells with a viral surrogate of recombinant GFP Ad5 in parallel with LV-CMV-GFP control. The best morphologic conditions were observed from the two lentiviruses co-infected cells, while single adenovirus infected cells underwent clear pathologic changes. Viral load of adenoviruses from LV-CMV-IFNγ or LV-CMV-GFP co-infected cell cultures was significantly lower than that from adenovirus alone infected cells (P=0.005-0.041), and the reduction of adenoviral load in the co-infected cells was 86% and 61%, respectively. Ad5 viral load from LV-CMV-IFNγ co-infected cells was significantly lower than that from LV-CMV-GFP co-infection (P=0.032), which suggested that IFNγ rather than GFP could further enhance the inhibition of Ad5 replication in the recombinant lentivirus co-infected cells. The results suggest that LV-CMV-IFNγ co-infection could significantly inhibit the target virus replication and might be a potential approach for alternative therapy of severe viral diseases.
探讨了在共感染腺病毒 5 型（Ad5）的细胞中培养慢病毒（LV）产生重组干扰素-γ（IFNγ）以抑制靶病毒的作用。在不同细胞系中系统评估了 CMV、EF1α 和 Ubiquitin 三个不同启动子在慢病毒内启动增强型绿色荧光蛋白（GFP）活性的能力，结果表明某些细胞系选择了最有利的启动子，可实现高水平的转基因表达。携带 CMV 启动子的重组 IFNγ 慢病毒（LV-CMV-IFNγ）与携带重组 GFP Ad5 的病毒替代物平行共感染 293A 细胞，同时作为 LV-CMV-GFP 对照。从共感染两种慢病毒的细胞中观察到最佳的形态条件，而单独感染腺病毒的细胞则发生明显的病变。LV-CMV-IFNγ 或 LV-CMV-GFP 共感染细胞培养物中腺病毒的病毒载量明显低于单独感染腺病毒的细胞（P=0.005-0.041），共感染细胞中腺病毒载量的降低分别为 86%和 61%。LV-CMV-IFNγ 共感染细胞中的 Ad5 病毒载量明显低于 LV-CMV-GFP 共感染（P=0.032），这表明 IFNγ 而不是 GFP 可以进一步增强重组慢病毒共感染细胞中 Ad5 复制的抑制作用。结果表明，LV-CMV-IFNγ 共感染可显著抑制靶病毒复制，可能是严重病毒病替代治疗的潜在方法。
