Preparation of quadri-subtype influenza virus-like particles using bovine immunodeficiency virus gag protein.
Preparation of quadri-subtype influenza virus-like particles using bovine immunodeficiency virus gag protein.
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DOI：
10.1016/j.virol.2015.10.007
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发表时间：
2016-01
期刊：
影响因子：
3.7
通讯作者：
Pushko P
中科院分区：
文献类型：
--
作者：
Tretyakova I;Hidajat R;Hamilton G;Horn N;Nickols B;Prather RO;Tumpey TM;Pushko P
由血凝素（HA）、神经氨酸酶（NA）和基质（M1）蛋白组成的流感VLP先前已用于免疫学和病毒学研究。在这里，我们证明了流感病毒VLP可以在Sf 9细胞中使用牛免疫缺陷病毒gag（Bgag）蛋白代替M1。我们发现Bgag可用于制备包括H1N1和H10 N8在内的几种流感亚型的VLP。此外，通过使用Bgag，我们制备了四亚型VLP，其在VLP内共表达来自禽源H5 N1、H7N9、H9 N2和H10 N8病毒的四种HA亚型。VLP显示血凝和神经氨酸酶活性，并与特异性抗血清反应。评价了四亚型VLP中每种HA亚型的含量和共定位。电子显微镜显示基于Bgag的VLP类似于直径为150-200 nm的流感病毒粒子。这是首次报道流感病毒VLP的四亚型设计和使用Bgag制备流感病毒VLP。
Influenza VLPs comprised of hemagglutinin (HA), neuraminidase (NA), and matrix (M1) proteins have been previously used for immunological and virological studies. Here we demonstrated that influenza VLPs can be made in Sf9 cells by using the bovine immunodeficiency virus gag (Bgag) protein in place of M1. We showed that Bgag can be used to prepare VLPs for several influenza subtypes including H1N1 and H10N8. Furthermore, by using Bgag, we prepared quadri-subtype VLPs, which co-expressed within the VLP the four HA subtypes derived from avian-origin H5N1, H7N9, H9N2 and H10N8 viruses. VLPs showed hemagglutination and neuraminidase activities and reacted with specific antisera. The content and co-localization of each HA subtype within the quadri-subtype VLP were evaluated. Electron microscopy showed that Bgag-based VLPs resembled influenza virions with the diameter of 150-200 nm. This is the first report of quadri-subtype design for influenza VLP and the use of Bgag for influenza VLP preparation.