Development of reverse genetics systems and investigation of host response antagonism and reassortment potential for Cache Valley and Kairi viruses, two emerging orthobunyaviruses of the Americas.
Development of reverse genetics systems and investigation of host response antagonism and reassortment potential for Cache Valley and Kairi viruses, two emerging orthobunyaviruses of the Americas.
复制标题
DOI：
10.1371/journal.pntd.0006884
复制
发表时间：
2018-10
影响因子：
3.8
通讯作者：
Kohl A
中科院分区：
文献类型：
--
作者：
Dunlop JI;Szemiel AM;Navarro A;Wilkie GS;Tong L;Modha S;Mair D;Sreenu VB;Da Silva Filipe A;Li P;Huang YS;Brennan B;Hughes J;Vanlandingham DL;Higgs S;Elliott RM;Kohl A
库什谷病毒（CVV）和凯里病毒（KRIV）等正布尼亚病毒是重要的动物病原体。CVV的周期性暴发导致北美农场羔羊大量死亡，而KRIV主要在南美洲和中美洲发现，地理范围几乎没有重叠。目前尚无针对这些病毒的疫苗或治疗方法。开发新型候选疫苗的一种方法是基于使用反向遗传学来产生引起免疫反应但不能恢复到完全毒力的减毒病毒。对两种病毒的全基因组进行测序，以获得最新的基因组序列信息。测序后，开发了CVV和KRIV的小基因组系统和反向遗传系统。CVV和KRIV的体外细胞宿主范围广，其中BHK-21细胞是适合病毒增殖和滴定的宿主细胞系。为了开发减毒病毒，破坏了NSs蛋白的开放阅读框。没有NSs蛋白表达的重组病毒诱导了I型干扰素（IFN）的产生，这表明NSs对两种病毒都具有IFN拮抗剂的功能，并且这种减毒病毒可以形成减毒病毒疫苗的基础。为了评估CVV和KRIV之间重组的潜力，这可能与重叠区域的疫苗接种活动有关，我们试图通过反向遗传学产生M段重组。我们无法获得这样的病毒，这表明这是一个不太可能发生的事件。Cache Valley病毒和Kairi病毒（CVV和KRIV；环布尼亚病毒科，正布尼亚病毒）是美洲重要的动物病原体。在这项研究中，我们开发了反向遗传学系统来研究和操纵这两种病毒的病毒基因组。病毒基因组发生突变以阻止NSs蛋白的表达，NSs蛋白是1型干扰素（IFN）系统的关键毒力因子和拮抗剂。在产生IFN的细胞系中进行的复制研究表明，与野生型病毒相比，携带nss缺失的病毒的生长速度较慢。相比之下，在缺乏IFN的细胞系中，两种病毒类型的生长具有可比性，突出了NSs在CVV和KRIV中作为IFN拮抗剂的作用。我们还通过遗传学研究证明，至少在这里测试的一种组合中，CVV和KRIV不太可能通过重组组合形成新的病毒，并提出这种重组病毒可能是合适的减毒活疫苗候选病毒。
Orthobunyaviruses such as Cache Valley virus (CVV) and Kairi virus (KRIV) are important animal pathogens. Periodic outbreaks of CVV have resulted in the significant loss of lambs on North American farms, whilst KRIV has mainly been detected in South and Central America with little overlap in geographical range. Vaccines or treatments for these viruses are unavailable. One approach to develop novel vaccine candidates is based on the use of reverse genetics to produce attenuated viruses that elicit immune responses but cannot revert to full virulence. The full genomes of both viruses were sequenced to obtain up to date genome sequence information. Following sequencing, minigenome systems and reverse genetics systems for both CVV and KRIV were developed. Both CVV and KRIV showed a wide in vitro cell host range, with BHK-21 cells a suitable host cell line for virus propagation and titration. To develop attenuated viruses, the open reading frames of the NSs proteins were disrupted. The recombinant viruses with no NSs protein expression induced the production of type I interferon (IFN), indicating that for both viruses NSs functions as an IFN antagonist and that such attenuated viruses could form the basis for attenuated viral vaccines. To assess the potential for reassortment between CVV and KRIV, which could be relevant during vaccination campaigns in areas of overlap, we attempted to produce M segment reassortants by reverse genetics. We were unable to obtain such viruses, suggesting that it is an unlikely event. Cache Valley and Kairi viruses (CVV and KRIV; Peribunyaviridae, Orthobunyavirus) are important animal pathogens of the Americas. In this study we developed reverse genetics systems to study and manipulate viral genomes of both viruses. Viral genomes were mutated to prevent the expression of the NSs protein, a key virulence factor and antagonist of the type 1 interferon (IFN) system. Replication studies in IFN producing cell lines showed slower growth of the NSs-deletion carrying viruses compared to wild type virus. In contrast, in IFN-deficient cell lines growth of both viral types was comparable, highlighting the role of NSs as an IFN antagonist in both CVV and KRIV. We also demonstrated using genetic studies that CVV and KRIV are unlikely to combine by reassortment to form novel viruses at least for one combination tested here, and propose that such recombinant viruses would be suitable live attenuated vaccine candidates.
登录
查看更多内容
影响因子：
4.4
作者：
Blomström AL;Gu Q;Barry G;Wilkie G;Skelton JK;Baird M;McFarlane M;Schnettler E;Elliott RM;Palmarini M;Kohl A
通讯作者：
Kohl A
影响因子：
2.1
作者：
Andreadis, Theodore G.;Armstrong, Philip M.;Main, Andrew J.
通讯作者：
Main, Andrew J.
DOI：
10.4269/ajtmh.1986.35.429
发表时间：
1986-03-01
影响因子：
3.3
作者：
CALISHER, CH;FRANCY, DB;MCLEAN, RG
通讯作者：
MCLEAN, RG
影响因子：
5.4
作者：
Blakqori, G;Weber, F
通讯作者：
Weber, F
影响因子：
2.1
作者：
Blackmore, CGM;Blackmore, MS;Grimstad, PR
通讯作者：
Grimstad, PR