Adenovirus-mediated RNA interference against foot-and-mouth disease virus infection both in vitro and in vivo
Adenovirus-mediated RNA interference against foot-and-mouth disease virus infection both in vitro and in vivo
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DOI：
10.1128/jvi.80.7.3559-3566.2006
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发表时间：
2006-04-01
影响因子：
5.4
通讯作者：
Zheng, ZX
中科院分区：
文献类型：
--
作者：
Chen, WZ;Liu, MQ;Zheng, ZX
口蹄疫病毒（FMDV）感染每年给畜牧业造成严重的经济损失。由于现有疫苗和抗病毒药物的有效性有限，因此需要制定新的策略。 RNA干扰（RNAi）是体外抑制病毒复制的有效手段。在这里，我们证明，用表达针对 FMDV 结构蛋白 1D (Ad5-NT21) 或聚合酶 3D (Ad5-POL) 的短发夹 RNA (shRNA) 的重组、复制缺陷型人 5 型腺病毒 (Ad5) 进行治疗，可以完全保护猪 IBRS-2 细胞免受同源 FMDV 感染，而只有 Ad5-POL 抑制异源 FMDV 复制。此外，这些 shRNA 的递送显着降低了豚鼠和猪对 FMDV 感染的易感性。五只豚鼠中的三只接种了 10(6) PFU 的 Ad5-POL，并在 24 小时后用 50 50% 感染剂量 (ID50) 的同源病毒进行攻击，它们免受疾病的主要临床表现：足部出现水疱。三只猪中的两只接种了各自含有 2 x 10(9) PFU 的 Ad5-NT21-Ad5-POL 混合物，并在 24 小时后用 100 1D 同源病毒进行攻击，免受主要临床疾病的影响，但用较高剂量的腺病毒混合物治疗不能促进对动物的保护。这种抑制是快速且特异的，因为用表达大肠杆菌半乳糖苷酶特异性 shRNA 的对照腺病毒构建体 (Ad5-LacZ) 处理没有显示出明显的抗病毒活性。我们的数据强调了 RNAi 技术在口蹄疫中的体内潜力。
Foot-and-mouth disease virus (FMDV) infection is responsible for the heavy economic losses in stockbreeding each year. Because of the limited effectiveness of existing vaccines and antiviral drugs, the development of new strategies is needed. RNA interference (RNAi) is an effective means of suppressing virus replication in vitro. Here we demonstrate that treatment with recombinant, replication-defective human adenovirus type 5 (Ad5) expressing short-hairpin RNAs (shRNAs) directed against either structural protein 1D (Ad5-NT21) or polymerase 3D (Ad5-POL) of FMDV totally protects swine IBRS-2 cells from homologous FMDV infection, whereas only Ad5-POL inhibits heterologous FMDV replication. Moreover, delivery of these shRNAs significantly reduces the susceptibility of guinea pigs and swine to FMDV infection. Three of five guinea pigs inoculated with 10(6) PFU of Ad5-POL and challenged 24 h later with 50 50% infectious doses (ID50) of homologous virus were protected from the major clinical manifestation of disease: the appearance of vesicles on the feet. Two of three swine inoculated with an Ad5-NT21-Ad5-POL mixture containing 2 x 10(9) PFU each and challenged 24 h later with 100 1D., of homologous virus were protected from the major clinical disease, but treatment with a higher dose of adenovirus mixture cannot promote protection of animals. The inhibition was rapid and specific because treatment with a control adenovirus construct (Ad5-LacZ) expressing Escherichia coli galactosidase-specific shRNA showed no marked antiviral activity. Our data highlight the in vivo potential of RNAi technology in the case of FMD.