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首页> 外文期刊>Molecular and Cellular Biology >Efficient and sustained gene expression in primary T lymphocytes and primary and cultured tumor cells mediated by adeno-associated virus plasmid DNA complexed to cationic liposomes.
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Efficient and sustained gene expression in primary T lymphocytes and primary and cultured tumor cells mediated by adeno-associated virus plasmid DNA complexed to cationic liposomes.

机译：通过与阳离子脂质体复合的腺相关病毒质粒DNA介导的原代T淋巴细胞以及原代和培养的肿瘤细胞中高效且持续的基因表达。

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摘要

We have used cationic liposomes to facilitate adeno-associated virus (AAV) plasmid transfections of primary and cultured cell types. AAV plasmid DNA complexed with liposomes showed levels of expression several fold higher than those of complexes with standard plasmids. In addition, long-term expression (> 30 days) of the gene, unlike the transient expression demonstrated by typical liposome-mediated transfection with standard plasmids, was observed. Southern analysis of chromosomal DNA further substantiated the hypothesis that the long-term expression was due to the presence of the transgene in the AAV plasmid-transfected group and not in the standard plasmid-transfected group. AAV plasmid-liposome complexes induced levels of transgene expression comparable to those obtained by recombinant AAV transduction. Primary breast, ovarian, and lung tumor cells were transfectable with the AAV plasmid DNA-liposome complexes. Transfected primary and cultured tumor cells were able to express transgene product even after lethal irradiation. High-level gene expression was also observed in freshly isolated CD3+, CD4+, and CD8+ T cells from normal human peripheral blood. Transfection efficiency ranged from 10 to 50% as assessed by intracellular interleukin-2 levels in interleukin-2-transfected cells. The ability to express transgenes in primary tumor and lymphoid cells may be applied toward tumor vaccine studies and protocols which may eventually permit highly specific modulation of the cellular immune response in cancer and AIDS.
机译：我们已经使用阳离子脂质体来促进原发性和培养细胞类型的腺相关病毒（AAV）质粒转染。与脂质体复合的AAV质粒DNA的表达水平比标准质粒复合物的表达水平高几倍。此外，观察到该基因的长期表达（> 30天），这与典型的脂质体介导的标准质粒转染所证明的瞬时表达不同。染色体DNA的Southern分析进一步证实了这一假设，即长期表达是由于在AAV质粒转染组而不是在标准质粒转染组中转基因的存在。 AAV质粒-脂质体复合物诱导的转基因表达水平与重组AAV转导所获得的水平相当。原发性乳腺，卵巢和肺部肿瘤细胞可用AAV质粒DNA-脂质体复合物转染。转染的原代和培养的肿瘤细胞即使在致死性照射后也能够表达转基因产物。在来自正常人外周血的新鲜分离的CD3 +，CD4 +和CD8 + T细胞中也观察到了高水平的基因表达。根据白介素2转染细胞的细胞内白介素2水平评估，转染效率为10％至5​​0％。在原发性肿瘤和淋巴样细胞中表达转基因的能力可用于肿瘤疫苗研究和方案，这些研究和方案可最终允许癌症和艾滋病中细胞免疫应答的高度特异性调节。

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