Institute of Medical Virology, Justus Liebig University, D-35392 Giessen, Germany.
J Virol. 2010 Feb;84(4):2122-33. doi: 10.1128/JVI.01668-09. Epub 2009 Dec 9.
A reassortant avian influenza virus (designated FPV NS GD), carrying the NS-segment of the highly pathogenic avian influenza virus (HPAIV) strain A/Goose/Guangdong/1/96 (GD; H5N1) in the genetic background of the HPAIV strain A/FPV/Rostock/34 (FPV; H7N1), was rescued by reverse genetics. Remarkably, in contrast to the recombinant wild-type FPV (rFPV), the reassortant virus was able to replicate more efficiently in different human cell lines and primary mouse epithelia cells without prior adaptation. Moreover, FPV NS GD caused disease and death in experimentally infected mice and was detected in mouse lungs; in contrast, rFPV was not able to replicate in mice effectively. These results indicated an altered host range and increased virulence. Furthermore FPV NS GD showed pronounced pathogenicity in chicken embryos. In an attempt to define the molecular basis for the apparent differences, we determined that NS1 proteins of the H5N1 and H7N1 strains bound the antiviral kinase PKR and the F2F3 domain of cleavage and polyadenylation specificity factor 30 (CPSF30) with comparable efficiencies in vitro. However, FPV NS GD infection resulted in (i) increased expression of NS1, (ii) faster and stronger PKR inhibition, and (iii) stronger beta interferon promoter inhibition than rFPV. Taken together, the results shed further light on the importance of the NS segment of an H5N1 strain for viral replication, molecular pathogenicity, and host range of HPAIVs and the possible consequences of a reassortment between naturally occurring H7 and H5 type HPAIVs.
一种重配的禽流感病毒(命名为 FPV NS GD),其 NS 节段来自高致病性禽流感病毒(HPAIV)株 A/鹅/广东/1/96(GD;H5N1),遗传背景为 HPAIV 株 A/FPV/Rostock/34(FPV;H7N1),通过反向遗传学技术被拯救。值得注意的是,与重组野生型 FPV(rFPV)相比,重配病毒能够更有效地在不同的人源细胞系和原代鼠上皮细胞中复制,而无需预先适应。此外,FPV NS GD 能够引起实验感染小鼠的疾病和死亡,并在小鼠肺部检测到;相比之下,rFPV 不能有效地在小鼠中复制。这些结果表明改变了宿主范围和增加了毒力。此外,FPV NS GD 在鸡胚中表现出明显的致病性。为了确定这些明显差异的分子基础,我们确定 H5N1 和 H7N1 株的 NS1 蛋白在体外以相当的效率结合抗病毒激酶 PKR 和切割和多聚腺苷酸化特异性因子 30(CPSF30)的 F2F3 结构域。然而,FPV NS GD 感染导致(i)NS1 蛋白表达增加,(ii)更快和更强的 PKR 抑制,以及(iii)更强的干扰素-β启动子抑制,这比 rFPV 更为明显。总之,这些结果进一步阐明了 H5N1 株的 NS 节段对 HPAIV 病毒复制、分子致病性和宿主范围的重要性,以及自然发生的 H7 和 H5 型 HPAIV 之间重配的可能后果。