ジャーナル: Nat Microbiol / 年: 2017 タイトル: Broadly protective murine monoclonal antibodies against influenza B virus target highly conserved neuraminidase epitopes. 著者: Teddy John Wohlbold / Kira A Podolsky / Veronika Chromikova / Ericka Kirkpatrick / Veronica Falconieri / Philip Meade / Fatima Amanat / Jessica Tan / Benjamin R tenOever / Gene S Tan / Sriram ...著者: Teddy John Wohlbold / Kira A Podolsky / Veronika Chromikova / Ericka Kirkpatrick / Veronica Falconieri / Philip Meade / Fatima Amanat / Jessica Tan / Benjamin R tenOever / Gene S Tan / Sriram Subramaniam / Peter Palese / Florian Krammer / 要旨: A substantial proportion of influenza-related childhood deaths are due to infection with influenza B viruses, which co-circulate in the human population as two antigenically distinct lineages defined ...A substantial proportion of influenza-related childhood deaths are due to infection with influenza B viruses, which co-circulate in the human population as two antigenically distinct lineages defined by the immunodominant receptor binding protein, haemagglutinin. While broadly cross-reactive, protective monoclonal antibodies against the haemagglutinin of influenza B viruses have been described, none targeting the neuraminidase, the second most abundant viral glycoprotein, have been reported. Here, we analyse a panel of five murine anti-neuraminidase monoclonal antibodies that demonstrate broad binding, neuraminidase inhibition, in vitro antibody-dependent cell-mediated cytotoxicity and in vivo protection against influenza B viruses belonging to both haemagglutinin lineages and spanning over 70 years of antigenic drift. Electron microscopic analysis of two neuraminidase-antibody complexes shows that the conserved neuraminidase epitopes are located on the head of the molecule and that they are distinct from the enzymatic active site. In the mouse model, one therapeutic dose of antibody 1F2 was more protective than the current standard of treatment, oseltamivir, given twice daily for six days.
超分子 #1: Influenza B neuraminidase bound to 4F11 Fab
超分子
名称: Influenza B neuraminidase bound to 4F11 Fab / タイプ: complex / ID: 1 / 親要素: 0
由来(天然)
生物種: Influenza B virus (B/Malaysia/2506/2004) (B型インフルエンザウイルス)
組換発現
生物種: Trichoplusia ni (イラクサキンウワバ)
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実験情報
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構造解析
手法
ネガティブ染色法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
緩衝液
pH: 7.3
染色
タイプ: NEGATIVE / 材質: Uranyl formate
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電子顕微鏡法
顕微鏡
FEI TECNAI 12
撮影
フィルム・検出器のモデル: GATAN ULTRASCAN 4000 (4k x 4k) 平均電子線量: 100.0 e/Å2
電子線
加速電圧: 120 kV / 電子線源: LAB6
電子光学系
照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD
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画像解析
最終 再構成
解像度のタイプ: BY AUTHOR / 解像度: 25.0 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: RELION (ver. 1.4) 詳細: The standard Fourier Shell Correlation-based measures for resolution are generally not very reliable for negative stain reconstructions because they tend to overestimate resolution and can ...詳細: The standard Fourier Shell Correlation-based measures for resolution are generally not very reliable for negative stain reconstructions because they tend to overestimate resolution and can result in spurious values that depend on data size rather than quality, a feature that is also true for cryo-EM reconstructions (some of these issues are discussed in Subramaniam et al, Curr Opin Str. Biol 41: 194-202 (2016)). In negative stain reconstructions, we therefore use a comparison of the experimentally obtained maps with maps computed from the fitted coordinates over a range of resolutions and use this to estimate resolution conservatively. The computed maps at 20 A and 25 A are easily comparable to an experimentally obtained map, which is why we used 25 A as an estimate for resolution. 使用した粒子像数: 13665