ジャーナル: JCI Insight / 年: 2016 タイトル: Recognition of influenza H3N2 variant virus by human neutralizing antibodies. 著者: Sandhya Bangaru / Travis Nieusma / Nurgun Kose / Natalie J Thornburg / Jessica A Finn / Bryan S Kaplan / Hannah G King / Vidisha Singh / Rebecca M Lampley / Gopal Sapparapu / Alberto Cisneros ...著者: Sandhya Bangaru / Travis Nieusma / Nurgun Kose / Natalie J Thornburg / Jessica A Finn / Bryan S Kaplan / Hannah G King / Vidisha Singh / Rebecca M Lampley / Gopal Sapparapu / Alberto Cisneros / Kathryn M Edwards / James C Slaughter / Srilatha Edupuganti / Lilin Lai / Juergen A Richt / Richard J Webby / Andrew B Ward / James E Crowe / 要旨: Since 2011, over 300 human cases of infection, especially in exposed children, with the influenza A H3N2 variant (H3N2v) virus that circulates in swine in the US have been reported. The structural ...Since 2011, over 300 human cases of infection, especially in exposed children, with the influenza A H3N2 variant (H3N2v) virus that circulates in swine in the US have been reported. The structural and genetic basis for the lack of protection against H3N2v induced by vaccines containing seasonal H3N2 antigens is poorly understood. We isolated 17 human monoclonal antibodies (mAbs) that neutralized H3N2v virus from subjects experimentally immunized with an H3N2v candidate vaccine. Six mAbs exhibited very potent neutralizing activity (IC < 200 ng/ml) against the H3N2v virus but not against current human H3N2 circulating strains. Fine epitope mapping and structural characterization of antigen-antibody complexes revealed that H3N2v specificity was attributable to amino acid polymorphisms in the 150-loop and the 190-helix antigenic sites on the hemagglutinin protein. H3N2v-specific antibodies also neutralized human H3N2 influenza strains naturally circulating between 1995 and 2005. These results reveal a high level of antigenic relatedness between the swine H3N2v virus and previously circulating human strains, consistent with the fact that early human H3 seasonal strains entered the porcine population in the 1990s and reentered the human population, where they had not been circulating, as H3N2v about a decade later. The data also explain the increased susceptibility to H3N2v viruses in young children, who lack prior exposure to human seasonal strains from the 1990s.
名称: H3v-47 Fab / タイプ: complex / ID: 2 / 親要素: 1 詳細: H3v-47 Fab in complex with A/Minnesota/11/2010 H3N2v HA and FI6v Fab
由来(天然)
生物種: Homo sapiens (ヒト)
組換発現
生物種: Homo sapiens (ヒト) / 組換細胞: human hybridoma cell line
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超分子 #3: FI6v Fab
超分子
名称: FI6v Fab / タイプ: complex / ID: 3 / 親要素: 1 詳細: Negative-stain EM reconstruction of H3v-47 Fab in complex with A/Minnesota/11/2010 H3N2v HA and FI6v Fab
由来(天然)
生物種: Human (ヒト) / 株: H3N2
組換発現
生物種: Spodoptera frugiperda (ツマジロクサヨトウ)
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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
染色
タイプ: NEGATIVE / 材質: Uranyl formate 詳細: The complex was diluted to 2ug/ml, applied to freshly glow discharged 400 mesh carbon coated copper grids, and negatively stained with 2% uranyl formate.
グリッド
モデル: EMS / 材質: COPPER / メッシュ: 400 / 支持フィルム - 材質: CELLULOSE ACETATE / 支持フィルム - トポロジー: CONTINUOUS / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 雰囲気: AIR / 詳細: Carbon was evaporated upon grid prior to use.
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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: TVIPS TEMCAM-F416 (4k x 4k) 平均電子線量: 24.7 e/Å2