Tretyakova Irina, Hidajat Rachmat, Hamilton Garrett, Horn Noah, Nickols Brian, Prather Raphael O, Tumpey Terrence M, Pushko Peter
Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA.
Influenza Division, Centers for Disease Control and Prevention, 1600 Clifton Road N.E., Atlanta, GA, USA.
Virology. 2016 Jan;487:163-71. doi: 10.1016/j.virol.2015.10.007. Epub 2015 Nov 2.
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 150-200nm. This is the first report of quadri-subtype design for influenza VLP and the use of Bgag for influenza VLP preparation.
由血凝素（HA）、神经氨酸酶（NA）和基质（M1）蛋白组成的流感病毒样颗粒（VLPs）此前已用于免疫学和病毒学研究。在此，我们证明了通过使用牛免疫缺陷病毒gag（Bgag）蛋白替代M1蛋白，可以在Sf9细胞中制备流感病毒样颗粒。我们表明，Bgag可用于制备包括H1N1和H10N8在内的几种流感亚型的病毒样颗粒。此外，通过使用Bgag，我们制备了四亚型病毒样颗粒，其在病毒样颗粒内共表达了源自禽源H5N1、H7N9、H9N2和H10N8病毒的四种HA亚型。病毒样颗粒表现出血凝和神经氨酸酶活性，并与特异性抗血清发生反应。评估了四亚型病毒样颗粒中每种HA亚型的含量和共定位情况。电子显微镜显示，基于Bgag的病毒样颗粒类似于直径为150 - 200nm的流感病毒粒子。这是关于流感病毒样颗粒四亚型设计以及使用Bgag制备流感病毒样颗粒的首次报道。
