Inhibition of HIV-1 by lentiviral vector-transduced siRNAs in T lymphocytes differentiated in SCID-hu mice and CD34+ progenitor cell-derived macrophages
Inhibition of HIV-1 by lentiviral vector-transduced siRNAs in T lymphocytes differentiated in SCID-hu mice and CD34+ progenitor cell-derived macrophages
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DOI：
10.1016/s1525-0016(03)00140-0
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发表时间：
2003-07-01
影响因子：
12.4
通讯作者：
Akkina, R
中科院分区：
文献类型：
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作者：
Banerjea, A;Li, MJ;Akkina, R
由小干扰RNA（siRNA）介导的RNA干扰现象是一种有效的基因沉默机制。最近的一些研究表明，使用这种方法可以抑制培养细胞中的HIV-1复制。为了取得进一步进展并利用这项技术在干细胞环境中进行HIV-1基因治疗，需要使用原代造血细胞进行体内研究。使用基于HIV的慢病毒载体，我们将抗Rev siRNA构建体引入CD 34（+）造血祖细胞中。在SCID-hu小鼠thy/liv移植物中，允许siRNA转导的祖细胞在体外成熟为巨噬细胞，在体内成熟为T细胞。获得表现出特征性表面标志物的表型正常的T细胞和巨噬细胞。在体外HIV-1的挑战，表达siRNA的巨噬细胞和T细胞与巨噬细胞嗜性和T细胞嗜性HIV-1，分别显示出显着的病毒耐药性。这些实验证明了通过慢病毒载体递送到造血干细胞中的siRNA对于未来体内应用的效用。
The phenomenon of RNA interference mediated by small interfering RNAs (siRNAs) is a potent gene-silencing mechanism. A number of recent studies demonstrated inhibition of HIV-1 replication in cultured cells using this approach. To make further progress and harness this technology for HIV-1 gene therapy in a stem cell setting, in vivo studies using primary hematopoietic cells are needed. Using an HIV-based lentiviral vector we introduced an anti-Rev siRNA construct into CD34(+) hematopoietic progenitor cells. The siRNA-transduced progenitor cells were allowed to mature into macrophages in vitro and T cells in vivo in SCID-hu mouse thy/liv grafts. Phenotypically normal T cells and macrophages displaying characteristic surface markers were obtained. In vitro HIV-1 challenge of the siRNA-expressing macrophages and T cells with macrophage-tropic and T-cell-tropic HIV-1, respectively, showed marked viral resistance. These experiments demonstrate the utility of siRNAs delivered into hematopoietic stem cells via lentiviral vectors for future in vivo applications.