Food Animal Health Research Program, Ohio Agricultural Research and Development Center, The Ohio State University, 1680 Madison Ave., Wooster, OH 44691, USA.
Vet Microbiol. 2011 Mar 24;148(2-4):175-82. doi: 10.1016/j.vetmic.2010.09.009. Epub 2010 Sep 16.
The H3N2 triple reassortant (TR) influenza viruses emerged in swine in 1998 and then in turkeys in 2003. It was then hypothesized that these viruses crossed the species barrier and transmitted from pigs to turkeys. In previous work we identified viruses with different transmission behavior between the two species, of which A/turkey/Ohio/313053/04 (TK04) transmitted both ways between swine and turkeys, and A/swine/North Carolina/03 (SW03) did not transmit either way between the two species. Utilizing the 12-plasmid reverse genetics (RG) system, we rescued two viruses (TK04 and SW03) with potentially different transmission behavior between pigs and turkeys. Single gene reassortants (SGR) were generated by switching the hemagglutinin (HA) or the neuraminidase (NA) genes between both viruses, and were evaluated for replication in vitro (pig and turkey tracheal/bronchial epithelial cells) and in vivo (pigs and turkeys). RG-created TK04 replicated more efficiently than SW03 in vitro and in vivo. Additionally, TK04 exhibited better binding affinity to plasma membrane preparations (PMP) from pig and turkey tracheal/bronchial epithelial cells compared to SW03. In study with SGR viruses, the HA protein was found to be essential for TK04 virus transmission amongst turkeys, but not sole factor contributing to the efficient replication of virus in turkeys and pigs. Such findings further highlight the polygenic nature of influenza virus pathogenesis.
1998 年，H3N2 三重重配（TR）流感病毒出现在猪群中，随后于 2003 年出现在火鸡中。人们推测这些病毒跨越了物种屏障，并从猪传播到火鸡。在之前的工作中，我们鉴定出了两种具有不同传播行为的病毒，其中 A/ turkey / Ohio / 313053 / 04（TK04）在猪和火鸡之间双向传播，而 A/ swine / North Carolina / 03（SW03）在这两个物种之间均不能传播。利用 12 质粒反向遗传（RG）系统，我们拯救了两种具有潜在不同传播行为的猪和火鸡之间的病毒（TK04 和 SW03）。通过在两种病毒之间交换血凝素（HA）或神经氨酸酶（NA）基因，产生了单基因重配体（SGR），并评估了它们在体外（猪和火鸡的气管/支气管上皮细胞）和体内（猪和火鸡）的复制情况。RG 产生的 TK04 在体外和体内的复制效率均高于 SW03。此外，与 SW03 相比，TK04 与猪和火鸡气管/支气管上皮细胞的质膜制剂（PMP）具有更好的结合亲和力。在 SGR 病毒研究中，发现 HA 蛋白对于 TK04 病毒在火鸡中的传播是必需的，但不是导致病毒在火鸡和猪中高效复制的唯一因素。这些发现进一步强调了流感病毒发病机制的多基因性质。
