Designs and Characterization of Subunit Ebola GP Vaccine Candidates: Implications for Immunogenicity.
Designs and Characterization of Subunit Ebola GP Vaccine Candidates: Implications for Immunogenicity.
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DOI：
10.3389/fimmu.2020.586595
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发表时间：
2020
影响因子：
7.3
通讯作者：
Spertini F
中科院分区：
文献类型：
--
作者：
Agnolon V;Kiseljak D;Wurm MJ;Wurm FM;Foissard C;Gallais F;Wehrle S;Muñoz-Fontela C;Bellanger L;Correia BE;Corradin G;Spertini F
埃博拉病毒（EBOV）幸存者的体液应答主要针对表面糖蛋白GP，抗GP中和抗体与针对EBOV感染的保护有关。为了通过疫苗接种引发保护性中和抗体，需要抗原的天然样构象。因此，我们在CHO细胞中工程化并表达了来自EBOV（扎伊尔种埃博拉病毒，Mayinga变体）的几种GP变体，包括可溶性GP ΔTM、粘蛋白样结构域缺失的GP ΔTM-ΔMUC以及两种具有C末端三聚基序的GP ΔTM-ΔMUC变体，以有利于其天然三聚体构象。包含三聚基序导致蛋白质模拟GP亚稳三聚体并显示出增加的稳定性。粘蛋白样结构域似乎不是保留GP蛋白的天然构象的关键，其去除暴露了几个中和表位，特别是在三聚体中。可溶性GP变体在假型转导测定中抑制mAb中和活性，进一步证实了蛋白质的结构完整性。有趣的是，三聚体GP，一种天然的GP复合物，对EBOV疾病幸存者中自然感染产生的抗体显示出更强的亲和力，而不是对接受ChAd 3-EBOZ疫苗的志愿者中产生的抗体。这些结果支持了我们的假设，即当使用GP抗原的天然样构象时，优先诱导中和抗体。我们开发的可溶性三聚体重组GP蛋白代表了开发针对EBOV和其他丝状病毒的预防性疫苗的新颖且有前途的策略。
The humoral responses of Ebola virus (EBOV) survivors mainly target the surface glycoprotein GP, and anti-GP neutralizing antibodies have been associated with protection against EBOV infection. In order to elicit protective neutralizing antibodies through vaccination a native-like conformation of the antigen is required. We therefore engineered and expressed in CHO cells several GP variants from EBOV (species Zaire ebolavirus, Mayinga variant), including a soluble GP ΔTM, a mucin-like domain-deleted GP ΔTM-ΔMUC, as well as two GP ΔTM-ΔMUC variants with C-terminal trimerization motifs in order to favor their native trimeric conformation. Inclusion of the trimerization motifs resulted in proteins mimicking GP metastable trimer and showing increased stability. The mucin-like domain appeared not to be critical for the retention of the native conformation of the GP protein, and its removal unmasked several neutralizing epitopes, especially in the trimers. The soluble GP variants inhibited mAbs neutralizing activity in a pseudotype transduction assay, further confirming the proteins’ structural integrity. Interestingly, the trimeric GPs, a native-like GP complex, showed stronger affinity for antibodies raised by natural infection in EBOV disease survivors rather than for antibodies raised in volunteers that received the ChAd3-EBOZ vaccine. These results support our hypothesis that neutralizing antibodies are preferentially induced when using a native-like conformation of the GP antigen. The soluble trimeric recombinant GP proteins we developed represent a novel and promising strategy to develop prophylactic vaccines against EBOV and other filoviruses.
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影响因子：
64.5
作者：
Flyak AI;Shen X;Murin CD;Turner HL;David JA;Fusco ML;Lampley R;Kose N;Ilinykh PA;Kuzmina N;Branchizio A;King H;Brown L;Bryan C;Davidson E;Doranz BJ;Slaughter JC;Sapparapu G;Klages C;Ksiazek TG;Saphire EO;Ward AB;Bukreyev A;Crowe JE Jr
通讯作者：
Crowe JE Jr
影响因子：
5.5
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影响因子：
2.9
作者：
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通讯作者：
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影响因子：
16.6
作者：
Janus BM;van Dyk N;Zhao X;Howell KA;Soto C;Aman MJ;Li Y;Fuerst TR;Ofek G
通讯作者：
Ofek G
影响因子：
14.9
作者：
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通讯作者：
Kardos J