The enhancement of RNAi against HIV in vitro and in vivo using H-2Kk protein as a sorting method
The enhancement of RNAi against HIV in vitro and in vivo using H-2Kk protein as a sorting method
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DOI：
10.1016/j.jviromet.2012.02.007
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发表时间：
2012-06-01
影响因子：
3.1
通讯作者：
Xia, Ningshao
中科院分区：
文献类型：
--
作者：
Gu, Ying;Hou, Wangheng;Xia, Ningshao
基因疗法为包括艾滋病毒/艾滋病在内的许多人类疾病提供了一种潜在的有效治疗方法。研究最多的基因传递系统之一是使用基于慢病毒的载体，它可以将基因传递到分裂和非分裂的细胞中。然而，感染效率低，阻碍了对其生物学功能的正确评价。本研究将一种表达短发夹rna （short hairpin rna, shRNAs）靶向HIV-1 vif/pol的重组慢病毒载体转导到多种细胞中。用MHC I类分子H-2K(k)作为标记物，通过免疫磁分选积累病毒转导细胞。转导细胞的体外测试显示，分选后的pbmc对HIV的抑制率为85%，而分选前的pbmc对HIV的抑制率为30%。此外，使用具有相同细胞富集处理方案的小鼠异种移植模型，在分选后的细胞中实现了约95%的HIV活性降低，而在预先分选的细胞中几乎没有。这些研究为积累病毒转导细胞提供了一种实用的方法，可用于评估各种shrna构建体的性能。(C) 2012 Elsevier BM。版权所有。
Gene therapy offers a potentially an effective treatment for many human diseases, including HIV/AIDS. One of the most studied gene delivery systems is the use of lentivirus based vectors, which can deliver genes into both dividing and nondividing cells. However, low infection efficiency represents an obstacle for proper evaluation of their biological function. In this study, a recombinant lentiviral vector which expressed short hairpin RNAs (shRNAs) targeted against the HIV-1 vif/pol was transduced into various cells. An MHC class I molecule, H-2K(k) was used as a marker to accumulate the virally transduced cells through immunomagnetic sorting. In vitro testing of transduced cells showed 85% suppression of HIV in post-sorted PBMCs compared to 30% in pre-sorted PBMCs. In additional, using a mouse xenotrans-plantation model with the same treatment protocol for cell enrichment, a > 95% decrease in HIV activity in post-sorted cells was achieved, as compared to nearly none in the pre-sorted cells. These studies offer a practical method to accumulate virally transduced cells, which can be applied to evaluate the performance of various shRNAs constructs. (C) 2012 Elsevier BM. All rights reserved.