Creanga Adrian, Gillespie Rebecca A, Fisher Brian E, Andrews Sarah F, Lederhofer Julia, Yap Christina, Hatch Liam, Stephens Tyler, Tsybovsky Yaroslav, Crank Michelle C, Ledgerwood Julie E, McDermott Adrian B, Mascola John R, Graham Barney S, Kanekiyo Masaru
Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Electron Microscopy Laboratory, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute, Frederick, MD, USA.
Nat Commun. 2021 Mar 19;12(1):1722. doi: 10.1038/s41467-021-21954-2.
Broadly neutralizing antibodies (bnAbs) have been developed as potential countermeasures for seasonal and pandemic influenza. Deep characterization of these bnAbs and polyclonal sera provides pivotal understanding for influenza immunity and informs effective vaccine design. However, conventional virus neutralization assays require high-containment laboratories and are difficult to standardize and roboticize. Here, we build a panel of engineered influenza viruses carrying a reporter gene to replace an essential viral gene, and develop an assay using the panel for in-depth profiling of neutralizing antibodies. Replication of these viruses is restricted to cells expressing the missing viral gene, allowing it to be manipulated in a biosafety level 2 environment. We generate the neutralization profile of 24 bnAbs using a 55-virus panel encompassing the near-complete diversity of human H1N1 and H3N2, as well as pandemic subtype viruses. Our system offers in-depth profiling of influenza immunity, including the antibodies against the hemagglutinin stem, a major target of universal influenza vaccines.
广泛中和抗体（bnAbs）已被开发为应对季节性流感和大流行性流感的潜在对策。对这些bnAbs和多克隆血清的深入表征为流感免疫提供了关键认识，并为有效的疫苗设计提供了依据。然而，传统的病毒中和试验需要高防护实验室，且难以标准化和自动化。在此，我们构建了一组携带报告基因以取代必需病毒基因的工程化流感病毒，并开发了一种使用该组病毒进行中和抗体深度分析的试验。这些病毒的复制仅限于表达缺失病毒基因的细胞，从而使其能够在生物安全2级环境中进行操作。我们使用一个包含人类H1N1和H3N2几乎全部多样性以及大流行亚型病毒的55种病毒的病毒组，生成了24种bnAbs的中和图谱。我们的系统提供了对流感免疫的深度分析，包括针对血凝素茎部的抗体，血凝素茎部是通用流感疫苗的主要靶点。
