Wu Yanhua, Zhang Guozhong, Li Yi, Jin Yi, Dale Rod, Sun Lun-Quan, Wang Ming
College of Veterinary Medicine, China Agriculture University, Beijing 100094, PR China.
Biochem Biophys Res Commun. 2008 Jan 11;365(2):369-74. doi: 10.1016/j.bbrc.2007.10.196. Epub 2007 Nov 9.
H5N1 avian influenza virus (AIV) has caused widespread infections in poultry and wild birds, and has the potential to emerge as a pandemic threat to human. In order to explore novel approaches to inhibiting highly pathogenic H5N1 influenza virus infection, we have developed short RNA oligonucleotides, specific for conserved regions of the non-structural protein gene (NS1) of AIV. In vitro the hemagglutination (HA) titers in RNA oligonucleotide-treated cells were at least 5-fold lower than that of the control. In vivo, the treatment with three doses of RNA oligonucleotides protected the infected chickens from H5N1 virus-induced death at a rate of up to 87.5%. Plaque assay and real-time PCR analysis showed a significant reduction of the PFU and viral RNA level in the lung tissues of the infected animals treated with the mixed RNA oligonucleotides targeting the NS1 gene. Together, our findings revealed that the RNA oligonucleotides targeting at the AIV NS1 gene could potently inhibit avian H5N1 influenza virus reproduction and present a rationale for the further development of the RNA oligonucleotides as prophylaxis and therapy for highly pathogenic H5N1 influenza virus infection in humans.
H5N1禽流感病毒（AIV）已在家禽和野鸟中引发广泛感染，并有成为对人类的大流行威胁的可能性。为了探索抑制高致病性H5N1流感病毒感染的新方法，我们开发了针对AIV非结构蛋白基因（NS1）保守区域的短RNA寡核苷酸。在体外，经RNA寡核苷酸处理的细胞中的血凝（HA）滴度比对照至少低5倍。在体内，用三剂RNA寡核苷酸进行治疗，以高达87.5%的比率保护受感染的鸡免于H5N1病毒诱导的死亡。空斑试验和实时PCR分析表明，在用靶向NS1基因的混合RNA寡核苷酸处理的受感染动物的肺组织中，空斑形成单位（PFU）和病毒RNA水平显著降低。总之，我们的研究结果表明，靶向AIV NS1基因的RNA寡核苷酸可有效抑制禽H5N1流感病毒的繁殖，并为进一步开发RNA寡核苷酸作为预防和治疗人类高致病性H5N1流感病毒感染提供了理论依据。
