Lentivirus-mediated RNA interference of DC-SIGN expression inhibits human immunodeficiency virus transmission from dendritic cells to T cells
Lentivirus-mediated RNA interference of DC-SIGN expression inhibits human immunodeficiency virus transmission from dendritic cells to T cells
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DOI：
10.1128/jvi.78.20.10848-10855.2004
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发表时间：
2004-10-01
影响因子：
5.4
通讯作者：
Piguet, V
中科院分区：
文献类型：
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作者：
Arrighi, JF;Pion, M;Piguet, V
在人类免疫缺陷病毒1型(HIV-1)感染的早期事件中，表达DC特异性细胞间黏附分子3-抓取非整合素(DC-SIGN)受体的未成熟树突状细胞(DC)在粘膜表面捕获少量HIV-1，并将病毒感染传播到淋巴结(22，34，45)中的CD4(+)T细胞。RNA干扰已成为深入了解基因功能的有力工具。为此，表达短发夹状RNA(ShRNA)的慢病毒载体可将小干扰RNA(SiRNA)送入哺乳动物细胞，是实现稳定基因沉默的有力工具。为了干扰DC-SIGN功能，我们开发了表达shRNA的慢病毒载体，能够有条件地抑制DC-SIGN的表达。利用不同的siRNA获得了人DC-SIGN和L-SIGN以及黑猩猩和恒河猴DC-SIGN抑制的选择性。抑制DC-SIGN的表达可抑制HIV-1gp120包膜糖蛋白与DC-SIGN转染体的黏附，以及HIV-1反式转移到靶细胞。此外，表达shRNA的慢病毒载体能够有效地抑制原代人DC的DC-SIGN表达。DC-SIGN阴性的DC不能增强HIV-1反式向T细胞传递HIV-1的感染性，这表明DC-SIGN受体在将感染性病毒颗粒从DC转移到T细胞中起重要作用。本系统在研究DC-SIGN在HIV发病机制中的作用以及该受体识别的其他病原体方面具有广泛的应用前景。
In the early events of human immunodeficiency virus type 1 (HIV-1) infection, immature dendritic cells (DCs) expressing the DC-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) receptor capture small amounts of HIV-1 on mucosal surfaces and spread viral infection to CD4(+) T cells in lymph nodes (22, 34, 45). RNA interference has emerged as a powerful tool to gain insight into gene function. For this purpose, lentiviral vectors that express short hairpin RNA (shRNA) for the delivery of small interfering RNA (siRNA) into mammalian cells represent a powerful tool to achieve stable gene silencing. In order to interfere with DC-SIGN function, we developed shRNA-expressing lentiviral vectors capable of conditionally suppressing DC-SIGN expression. Selectivity of inhibition of human DC-SIGN and L-SIGN and chimpanzee and rhesus macaque DC-SIGN was obtained by using distinct siRNAs. Suppression of DC-SIGN expression inhibited the attachment of the gp120 envelope glycoprotein of HIV-1 to DC-SIGN transfectants, as well as transfer of HIV-1 to target cells in trans. Furthermore, shRNA-expressing lentiviral vectors were capable of efficiently suppressing DC-SIGN expression in primary human DCs. DC-SIGN-negative DCs were unable to enhance transfer of HIV-1 infectivity to T cells in trans, demonstrating an essential role for the DC-SIGN receptor in transferring infectious viral particles from DCs to T cells. The present system should have broad applications for studying the function of DC-SIGN in the pathogenesis of HIV as well as other pathogens also recognized by this receptor.