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基本情報
登録情報 | データベース: EMDB / ID: EMD-6077 | |||||||||
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タイトル | Characterization of a broadly neutralizing monoclonal antibody that targets the fusion domain of group 2 influenza A virus hemagglutinin | |||||||||
![]() | Negative stain reconstruction of Fab 9H10 bound to hemagglutinin (H3) from Influenza A/Hong Kong/1/1968 (H3N2), initiated with common lines, refined with projection matching against single particles, under C3 symmetry | |||||||||
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![]() | group 2 influenza A virus / broadly neutralizing antibody | |||||||||
生物種 | ![]() ![]() ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / ネガティブ染色法 / 解像度: 24.6 Å | |||||||||
![]() | Tan GS / Lee PS / Hoffman RMB / Mazel-Sanchez B / Krammer F / Leon PE / Ward AB / Wilson IA / Palese P | |||||||||
![]() | ![]() タイトル: Characterization of a broadly neutralizing monoclonal antibody that targets the fusion domain of group 2 influenza A virus hemagglutinin. 著者: Gene S Tan / Peter S Lee / Ryan M B Hoffman / Beryl Mazel-Sanchez / Florian Krammer / Paul E Leon / Andrew B Ward / Ian A Wilson / Peter Palese / ![]() 要旨: Due to continuous changes to its antigenic regions, influenza viruses can evade immune detection and cause a significant amount of morbidity and mortality around the world. Influenza vaccinations can ...Due to continuous changes to its antigenic regions, influenza viruses can evade immune detection and cause a significant amount of morbidity and mortality around the world. Influenza vaccinations can protect against disease but must be annually reformulated to match the current circulating strains. In the development of a broad-spectrum influenza vaccine, the elucidation of conserved epitopes is paramount. To this end, we designed an immunization strategy in mice to boost the humoral response against conserved regions of the hemagglutinin (HA) glycoprotein. Of note, generation and identification of broadly neutralizing antibodies that target group 2 HAs are rare and thus far have yielded only a few monoclonal antibodies (MAbs). Here, we demonstrate that mouse MAb 9H10 has broad and potent in vitro neutralizing activity against H3 and H10 group 2 influenza A subtypes. In the mouse model, MAb 9H10 protects mice against two divergent mouse-adapted H3N2 strains, in both pre- and postexposure administration regimens. In vitro and cell-free assays suggest that MAb 9H10 inhibits viral replication by blocking HA-dependent fusion of the viral and endosomal membranes early in the replication cycle and by disrupting viral particle egress in the late stage of infection. Interestingly, electron microscopy reconstructions of MAb 9H10 bound to the HA reveal that it binds a similar binding footprint to MAbs CR8020 and CR8043. IMPORTANCE: The influenza hemagglutinin is the major antigenic target of the humoral immune response. However, due to continuous antigenic changes that occur on the surface of this glycoprotein, ...IMPORTANCE: The influenza hemagglutinin is the major antigenic target of the humoral immune response. However, due to continuous antigenic changes that occur on the surface of this glycoprotein, influenza viruses can escape the immune system and cause significant disease to the host. Toward the development of broad-spectrum therapeutics and vaccines against influenza virus, elucidation of conserved regions of influenza viruses is crucial. Thus, defining these types of epitopes through the generation and characterization of broadly neutralizing monoclonal antibodies (MAbs) can greatly assist others in highlighting conserved regions of hemagglutinin. Here, we demonstrate that MAb 9H10 that targets the hemagglutinin stalk has broadly neutralizing activity against group 2 influenza A viruses in vitro and in vivo. | |||||||||
履歴 |
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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マップデータ | ![]() | 959.2 KB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 10.8 KB 10.8 KB | 表示 表示 | ![]() |
画像 | ![]() | 79.3 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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リンク
EMDBのページ | ![]() ![]() |
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マップ
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注釈 | Negative stain reconstruction of Fab 9H10 bound to hemagglutinin (H3) from Influenza A/Hong Kong/1/1968 (H3N2), initiated with common lines, refined with projection matching against single particles, under C3 symmetry | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 6.15 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
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試料の構成要素
-全体 : Fab 9H10 bound to hemagglutinin (H3) from Influenza A/Hong Kong/1...
全体 | 名称: Fab 9H10 bound to hemagglutinin (H3) from Influenza A/Hong Kong/1/1968 (H3N2) |
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要素 |
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-超分子 #1000: Fab 9H10 bound to hemagglutinin (H3) from Influenza A/Hong Kong/1...
超分子 | 名称: Fab 9H10 bound to hemagglutinin (H3) from Influenza A/Hong Kong/1/1968 (H3N2) タイプ: sample / ID: 1000 集合状態: one hemagglutinin homotrimer binds to three 9H10 Fabs Number unique components: 2 |
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分子量 | 理論値: 300 KDa |
-分子 #1: hemagglutinin
分子 | 名称: hemagglutinin / タイプ: protein_or_peptide / ID: 1 / コピー数: 3 / 集合状態: trimer / 組換発現: Yes / データベース: NCBI |
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由来(天然) | 生物種: ![]() ![]() 株: A/Hong Kong/1/1968 (H3N2) / 別称: flu virus |
分子量 | 理論値: 150 KDa |
-分子 #2: Fab of monoclonal antibody 9H10
分子 | 名称: Fab of monoclonal antibody 9H10 / タイプ: protein_or_peptide / ID: 2 / Name.synonym: antigen-binding fragment / コピー数: 3 / 集合状態: monomer / 組換発現: Yes / データベース: NCBI |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 50 KDa |
-実験情報
-構造解析
手法 | ネガティブ染色法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.01 mg/mL |
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緩衝液 | pH: 7.4 / 詳細: 150 mM NaCl, 20 mM Tris |
染色 | タイプ: NEGATIVE 詳細: Protein adsorbed for 20 seconds, blotted, stained with 2% uranyl formate for 20 seconds, and blotted again. |
グリッド | 詳細: 400 mesh copper grid coated with nitrocellulose and a thin layer of carbon |
凍結 | 凍結剤: NONE / 装置: OTHER |
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電子顕微鏡法
顕微鏡 | FEI TECNAI SPIRIT |
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温度 | 平均: 298 K |
アライメント法 | Legacy - 非点収差: Objective lens astigmatism monitored/corrected using continuously acquired FFTs at 52000X. |
日付 | 2014年2月1日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: TVIPS TEMCAM-F416 (4k x 4k) 実像数: 283 |
Tilt angle max | 0 |
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 / 最小 デフォーカス(公称値): 1.0 µm / 倍率(公称値): 52000 |
試料ステージ | 試料ホルダーモデル: SIDE ENTRY, EUCENTRIC / Tilt angle min: -55 |
実験機器 | ![]() モデル: Tecnai Spirit / 画像提供: FEI Company |
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画像解析
詳細 | Initial model determined by common lines from reference-free class averages. Final model refined against single particles using projection matching. |
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最終 再構成 | アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 24.6 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: EMAN2, Sparx / 使用した粒子像数: 4791 |