Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100021, China.
Int Immunopharmacol. 2011 Dec;11(12):2057-61. doi: 10.1016/j.intimp.2011.08.019. Epub 2011 Sep 18.
The H5N1 avian influenza virus (AIV) causes widespread infections in bird and human respiratory tracts, and vaccines and drug therapy are limited in their effectiveness. Recent studies of AIV structures have been published and provide new targets for designing antiviral drugs such as antisense oligonucleotides (AS ODNs), which effectively inhibit gene replication. In this study, we designed and synthesized three AS ODNs (NP267, NP628, NP749) that were specific for the RNA binding region of nucleoprotein (NP) based on AIV structure. Results showed that all three AS ODNs could inhibit viral replication in MDCK cells. The NP628 showed the best antiviral effect of all through viral titers, quantitative RT-PCR and indirect immunofluorescence (IFA) assays. In addition, the liposome mediated NP628 could partially protect the mice from a lethal H5N1 influenza virus challenge. Moreover, the NP628 group had a lower viral titer and lung index in the infected mice when compared with the viral control. Our results showed that AS ODN targeting of the AIV NP gene could potently inhibit AIV H5N1 reproduction, thus, formulating a candidate for an emergent therapeutic drug for the pathogenic H5N1 influenza virus infection.
H5N1 禽流感病毒（AIV）在鸟类和人类呼吸道中引起广泛感染，疫苗和药物治疗的效果有限。最近发表的 AIV 结构研究为设计抗病毒药物提供了新的靶点，例如反义寡核苷酸（AS ODN），它可以有效地抑制基因复制。在这项研究中，我们根据 AIV 结构设计并合成了三种针对核蛋白（NP）RNA 结合区域的 AS ODN（NP267、NP628、NP749）。结果表明，这三种 AS ODN 都可以抑制 MDCK 细胞中的病毒复制。通过病毒滴度、定量 RT-PCR 和间接免疫荧光（IFA）检测，NP628 显示出最佳的抗病毒效果。此外，脂质体介导的 NP628 可以部分保护小鼠免受致死性 H5N1 流感病毒的攻击。此外，与病毒对照组相比，感染 NP628 组的小鼠病毒滴度和肺指数均较低。我们的研究结果表明，针对 AIV NP 基因的 AS ODN 可以有效抑制 AIV H5N1 的复制，因此，为治疗致病性 H5N1 流感病毒感染提供了一种候选药物。
