ジャーナル: Biotechnol Bioeng / 年: 2023 タイトル: Mimotope discovery as a tool to design a vaccine against Zika and dengue viruses. 著者: Esmeralda Cuevas-Juárez / Arturo Liñan-Torres / Carolina Hernández / Mykhailo Kopylov / Clint S Potter / Bridget Carragher / Octavio T Ramírez / Laura A Palomares / 要旨: Vaccine development against dengue virus is challenging because of the antibody-dependent enhancement of infection (ADE), which causes severe disease. Consecutive infections by Zika (ZIKV) and/or ...Vaccine development against dengue virus is challenging because of the antibody-dependent enhancement of infection (ADE), which causes severe disease. Consecutive infections by Zika (ZIKV) and/or dengue viruses (DENV), or vaccination can predispose to ADE. Current vaccines and vaccine candidates contain the complete envelope viral protein, with epitopes that can raise antibodies causing ADE. We used the envelope dimer epitope (EDE), which induces neutralizing antibodies that do not elicit ADE, to design a vaccine against both flaviviruses. However, EDE is a discontinuous quaternary epitope that cannot be isolated from the E protein without other epitopes. Utilizing phage display, we selected three peptides that mimic the EDE. Free mimotopes were disordered and did not elicit an immune response. After their display on adeno-associated virus (AAV) capsids (VLP), they recovered their structure and were recognized by an EDE-specific antibody. Characterization by cryo-EM and enzyme-linked immunosorbent assay confirmed the correct display of a mimotope on the surface of the AAV VLP and its recognition by the specific antibody. Immunization with the AAV VLP displaying one of the mimotopes induced antibodies that recognized ZIKV and DENV. This work provides the basis for developing a Zika and dengue virus vaccine candidate that will not induce ADE.
名称: adeno-associated virus 2 / タイプ: virus / ID: 1 / 親要素: 0 詳細: Produced using the Baculovirus expression vector system and the particles were purified from the cellular pellet using an iodixanol gradient NCBI-ID: 10804 / 生物種: adeno-associated virus 2 / ウイルスタイプ: VIRUS-LIKE PARTICLE / ウイルス・単離状態: SEROTYPE / ウイルス・エンベロープ: No / ウイルス・中空状態: Yes
-
実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
濃度
1.0 mg/mL
緩衝液
pH: 7.5
凍結
凍結剤: ETHANE / チャンバー内湿度: 90 % / 装置: LEICA EM GP
-
電子顕微鏡法
顕微鏡
TFS KRIOS
撮影
フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 撮影したグリッド数: 1 / 実像数: 3727 / 平均露光時間: 2.0 sec. / 平均電子線量: 47.77 e/Å2
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
C2レンズ絞り径: 100.0 µm / 照射モード: OTHER / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 2.5 µm / 最小 デフォーカス(公称値): 0.8 µm