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首页> 外文期刊>Journal of interferon and cytokine research: The official journal of the International Society for Interferon and Cytokine Research >Transduction and utility of the granulocyte-macrophage colony-stimulating factor gene into monocytes and dendritic cells by adeno-associated virus.
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Transduction and utility of the granulocyte-macrophage colony-stimulating factor gene into monocytes and dendritic cells by adeno-associated virus.

机译：腺相关病毒将粒细胞-巨噬细胞集落刺激因子基因转导和用于单核细胞和树突状细胞。

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The genetic manipulation of antigen-presenting dendritic cells (DC) offers promise for stimulating the immune response, in particular for anticancer and antiviral protocols. As adeno-associated virus (AAV) has shown promise as a gene delivery vector for transducing a variety of hematopoietic cell types, we have investigated AAV's ability to genetically alter DC. In this analysis, we modified the standard granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) treatment of adherent monocytes to generate DC. In our protocol, adherent monocytes were first infected with an AAV/GM-CSF/Neo vector, and the addition of IL-4 was delayed for 2 days to allow for a brief period of monocyte proliferation. AAV-mediated transduction of the GM-CSF and Neo genes into monocytes/DC precursors was demonstrated by G418 selection, GM-CSF secretion, GM-CSF RNA expression (reverse transcriptase-polymerase chain reaction amplification [RT-PCR]), and cell proliferation. Cells resulting from infection with AAV/GM-CSF/Neo virus, and subsequent IL-4 and tumor necrosis factor-alpha (TNF-alpha) treatment, displayed multiple classic markers consistent with mature DC. Finally, chromosomal integration of the AAV vector was also demonstrated in sorted CD83+ DC. These data strongly suggest that AAV vectors will be useful for the genetic manipulation of DC and suggest that the transduction of the GM-CSF gene was able to fully replace the need for exogenous GM-CSF in the production of mature DC.
机译：抗原呈递树突状细胞（DC）的基因操作为刺激免疫应答提供了希望，特别是对于抗癌和抗病毒方案。腺伴随病毒（AAV）已显示出有望作为转导多种造血细胞类型的基因传递载体，因此我们研究了AAV遗传改变DC的能力。在此分析中，我们修改了标准的粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素4（IL-4）处理粘附的单核细胞以产生DC。在我们的方案中，首先用AAV / GM-CSF / Neo载体感染粘附的单核细胞，并将IL-4的添加延迟2天，以允许短暂的单核细胞增殖期。通过G418选择，GM-CSF分泌，GM-CSF RNA表达（逆转录-聚合酶链反应扩增[RT-PCR]）和细胞证明了AAV介导的GM-CSF和Neo基因转入单核细胞/ DC前体增殖。由AAV / GM-CSF / Neo病毒感染以及随后的IL-4和肿瘤坏死因子-α（TNF-alpha）处理产生的细胞显示出与成熟DC一致的多个经典标记。最后，在分选的CD83 + DC中也证明了AAV载体的染色体整合。这些数据强烈表明，AAV载体将对DC的基因操作有用，并表明GM-CSF基因的转导能够完全替代成熟DC生产中对外源GM-CSF的需求。

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