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PA-X protein contributes to virulence of triple-reassortant H1N2 influenza virus by suppressing early immune responses in swine

Virology. 2017; 
Guanlong Xua,b, Xuxiao Zhanga, Qinfang Liuc, Guoxia Bingd, Zhe Hua, Honglei Suna, Xin Xionga, Ming Jianga, Qiming Hea, Yu Wanga, Juan Pua, Xin Guoa, Hanchun Yanga, Jinhua Liua, Yipeng Suna
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Peptide Synthesis The protein samples derived from cell lysates were heated at 100 °C for 10 min and then separated on 10% sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) gels and transferred to polyvinylidene difluoride (PVDF) membranes (Bio-Rad), and subsequently incubated with primary antibodies, which were specific for influenza A virus PA (diluted 1:3000, GeneTex, USA), influenza A virus PA-X [diluted 1:2000, polyclonal rabbit antiserum against the PA-X derived peptide (CAGLPTKVSHRTSPA), Genscript, China), β-actin (diluted 1:5000, Beyotime, China). The secondary antibodies used were either horseradish peroxidase (HRP)-conjugated anti-mouse antibody or HRP-conjugated anti-rabbit antibody (diluted 1:10,000 Jackson ImmunoResearch USA), as appropriate. Get A Quote

摘要

PA-X protein Swine influenza virus Virulence Immune responses

关键词

Previous studies have identified a functional role of PA-X for influenza viruses in mice and avian species; however, its role in swine remains unknown. Toward this, we constructed PA-X deficient virus (Sw-FS) in the background of a Triple-reassortment (TR) H1N2 swine influenza virus (SIV) to assess the impact of PA-X in viral virulence in pigs. Expression of PA-X in TR H1N2 SIV enhanced viral replication and host protein synthesis shutoff, and inhibited the mRNA levels of type I IFNs and proinflammatory cytokines in porcine cells. A delay of proinflammatory responses was observed in lungs of pigs infected by wild type SIV (Sw-WT) compared to Sw-FS. Furthermore, Sw-WT virus replicated and transmitted more efficiently than Sw-FS in pigs. These results highlight the importance of PA-X in the moderation of virulence and immune responses of TR SIV in swine, which indicated that PA-X is a pro-virulence factor in TR SIV in pigs.
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