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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus
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Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus

机译：使用GFP报告病毒分析小鼠中流感病毒感染的体内动力学

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摘要

Influenza A virus is being extensively studied because of its major impact on human and animal health. However, the dynamics of influenza virus infection and the cell types infected in vivo are poorly understood. These characteristics are challenging to determine, partly because there is no efficient replication-competent virus expressing an easily traceable reporter gene. Here, we report the generation of a recombinant influenza virus carrying a GFP reporter gene in the NS segment (NS1-GFP virus). Although attenuated when compared with wild-type virus, the NS1-GFP virus replicates efficiently in murine lungs and shows pathogenicity in mice. Using whole-organ imaging and flow cytometry, we have tracked the dynamics of influenza virus infection progression in mice. Imaging of murine lungs shows that infection starts in the respiratory tract in areas close to large conducting airways and later spreads to deeper sections of the lungs. In addition to epithelial cells, we found GFP-positive antigen-presenting cells, such as CD11b~+CD11c~-, CD11b~-CD11c~+, and CD11b~+CD11c~+, as early as 24 h after intranasal infection. In addition, a significant proportion of NK and B cells were GFP positive, suggesting active infection of these cells. We next tested the effects of the influenza virus inhibitors oseltamivir and amantadine on the kinetics of in vivo infection progression. Treatment with oseltamivir dramatically reduced influenza infection in all cell types, whereas, surprisingly, amantadine treatment more efficiently blocked infection in B and NK cells. Our results demonstrate high levels of immune cells harboring influenza virus antigen during viral infection and cell-type-specific effects upon treatment with antiviral agents, opening additional avenues of research in the influenza virus field.
机译：由于甲型流感病毒对人类和动物健康具有重大影响，因此正在广泛研究。但是，人们对流感病毒感染的动力学和体内感染的细胞类型知之甚少。确定这些特征具有挑战性，部分原因是没有有效的具有复制能力的病毒表达易于追踪的报道基因。在此，我们报告了在NS片段中携带GFP报告基因的重组流感病毒的产生（NS1-GFP病毒）。尽管与野生型病毒相比减毒，但NS1-GFP病毒在鼠肺中有效复制，并在小鼠中显示出致病性。使用全器官成像和流式细胞仪，我们已经跟踪了小鼠中流感病毒感染进展的动态。鼠肺成像显示，感染始于呼吸道，靠近大型导气管，随后扩散至肺深处。除上皮细胞外，我们还发现鼻腔感染后24 h内CD11b〜+ CD11c〜-，CD11b〜-CD11c〜+和CD11b〜+ CD11c〜+等GFP阳性抗原呈递细胞。另外，相当大比例的NK和B细胞为GFP阳性，表明这些细胞被主动感染。接下来，我们测试了流感病毒抑制剂奥司他韦和金刚烷胺对体内感染进展动力学的影响。用奥司他韦治疗可以显着减少所有细胞类型的流感感染，而令人惊讶的是，金刚烷胺治疗可以更有效地阻断B细胞和NK细胞的感染。我们的结果表明，在病毒感染期间携带流感病毒抗原的免疫细胞水平高，并且在用抗病毒剂治疗后具有细胞类型特异性作用，这为流感病毒领域的研究开辟了新途径。

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  • 作者

    Balaji Manicassamy; Santhakumar Manicassamy; Alan Belicha-Villanueva; Giuseppe Pisanelli; Bali Pulendran; Adolfo Garcia-Sastre;

  • 作者单位

    Department of Microbiology, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029 Global Health and Emerging Pathogens Institute, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029;

    rnEmory Vaccine Center, Emory University, Atlanta, GA 30329;

    rnDepartment of Microbiology, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029 Global Health and Emerging Pathogens Institute, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029;

    rnDepartment of Microbiology, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029 Department of Pathology and Animal Health, School of Biotechnological Sciences, University of Naples 'Federico II', 80137 Naples, Italy;

    rnEmory Vaccine Center, Emory University, Atlanta, GA 30329;

    rnDepartment of Microbiology, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029 Global Health and Emerging Pathogens Institute, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029 Department of Medicine, Division of Infectious Diseases, Mount Sinai School of Medicine, New York, NY 10029;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    antivirals; pathogenesis; recombinant influenza virus; GFP virus; cell tropism;

    机译：抗病毒药发病;重组流感病毒;GFP病毒;细胞趋向性;

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