Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus.
The Pneumoviridae family of viruses includes human metapneumovirus (HMPV) and respiratory syncytial virus (RSV). The closely related Paramyxoviridae family includes parainfluenza viruses (PIVs). These three viral pathogens cause acute respiratory tract infections with substantial disease burden in t...
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Public Library of Science (PLoS)
2023-09-01
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| Series: | PLoS Pathogens |
| Online Access: | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1011584&type=printable |
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| author | Li Ou Steven J Chen I-Ting Teng Lijuan Yang Baoshan Zhang Tongqing Zhou Andrea Biju Cheng Cheng Wing-Pui Kong Nicholas C Morano Erik-Stephane D Stancofski John-Paul Todd Yaroslav Tsybovsky Shuishu Wang Cheng-Yan Zheng John R Mascola Lawrence Shapiro Ruth A Woodward Ursula J Buchholz Peter D Kwong |
| author_facet | Li Ou Steven J Chen I-Ting Teng Lijuan Yang Baoshan Zhang Tongqing Zhou Andrea Biju Cheng Cheng Wing-Pui Kong Nicholas C Morano Erik-Stephane D Stancofski John-Paul Todd Yaroslav Tsybovsky Shuishu Wang Cheng-Yan Zheng John R Mascola Lawrence Shapiro Ruth A Woodward Ursula J Buchholz Peter D Kwong |
| author_sort | Li Ou |
| collection | DOAJ |
| description | The Pneumoviridae family of viruses includes human metapneumovirus (HMPV) and respiratory syncytial virus (RSV). The closely related Paramyxoviridae family includes parainfluenza viruses (PIVs). These three viral pathogens cause acute respiratory tract infections with substantial disease burden in the young, the elderly, and the immune-compromised. While promising subunit vaccines are being developed with prefusion-stabilized forms of the fusion glycoproteins (Fs) of RSV and PIVs, for which neutralizing titers elicited by the prefusion (pre-F) conformation of F are much higher than for the postfusion (post-F) conformation, with HMPV, pre-F and post-F immunogens described thus far elicit similar neutralizing responses, and it has been unclear which conformation, pre-F or post-F, would be the most effective HMPV F-vaccine immunogen. Here, we investigate the impact of further stabilizing HMPV F in the pre-F state. We replaced the furin-cleavage site with a flexible linker, creating a single chain F that yielded increased amounts of pre-F stabilized trimers, enabling the generation and assessment of F trimers stabilized by multiple disulfide bonds. Introduced prolines could increase both expression yields and antigenic recognition by the pre-F specific antibody, MPE8. The cryo-EM structure of a triple disulfide-stabilized pre-F trimer with the variable region of antibody MPE8 at 3.25-Å resolution confirmed the formation of designed disulfides and provided structural details on the MPE8 interface. Immunogenicity assessments in naïve mice showed the triple disulfide-stabilized pre-F trimer could elicit high titer neutralization, >10-fold higher than elicited by post-F. Immunogenicity assessments in pre-exposed rhesus macaques showed the triple disulfide-stabilized pre-F could recall high neutralizing titers after a single immunization, with little discrimination in the recall response between pre-F and post-F immunogens. However, the triple disulfide-stabilized pre-F adsorbed HMPV-directed responses from commercially available pooled human immunoglobulin more fully than post-F. Collectively, these results suggest single-chain triple disulfide-stabilized pre-F trimers to be promising HMPV-vaccine antigens. |
| format | Article |
| id | doaj-art-fd9c2d280940481c856c94d7c813316a |
| institution | Kabale University |
| issn | 1553-7366 1553-7374 |
| language | English |
| publishDate | 2023-09-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj-art-fd9c2d280940481c856c94d7c813316a2025-08-20T03:25:46ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742023-09-01199e101158410.1371/journal.ppat.1011584Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus.Li OuSteven J ChenI-Ting TengLijuan YangBaoshan ZhangTongqing ZhouAndrea BijuCheng ChengWing-Pui KongNicholas C MoranoErik-Stephane D StancofskiJohn-Paul ToddYaroslav TsybovskyShuishu WangCheng-Yan ZhengJohn R MascolaLawrence ShapiroRuth A WoodwardUrsula J BuchholzPeter D KwongThe Pneumoviridae family of viruses includes human metapneumovirus (HMPV) and respiratory syncytial virus (RSV). The closely related Paramyxoviridae family includes parainfluenza viruses (PIVs). These three viral pathogens cause acute respiratory tract infections with substantial disease burden in the young, the elderly, and the immune-compromised. While promising subunit vaccines are being developed with prefusion-stabilized forms of the fusion glycoproteins (Fs) of RSV and PIVs, for which neutralizing titers elicited by the prefusion (pre-F) conformation of F are much higher than for the postfusion (post-F) conformation, with HMPV, pre-F and post-F immunogens described thus far elicit similar neutralizing responses, and it has been unclear which conformation, pre-F or post-F, would be the most effective HMPV F-vaccine immunogen. Here, we investigate the impact of further stabilizing HMPV F in the pre-F state. We replaced the furin-cleavage site with a flexible linker, creating a single chain F that yielded increased amounts of pre-F stabilized trimers, enabling the generation and assessment of F trimers stabilized by multiple disulfide bonds. Introduced prolines could increase both expression yields and antigenic recognition by the pre-F specific antibody, MPE8. The cryo-EM structure of a triple disulfide-stabilized pre-F trimer with the variable region of antibody MPE8 at 3.25-Å resolution confirmed the formation of designed disulfides and provided structural details on the MPE8 interface. Immunogenicity assessments in naïve mice showed the triple disulfide-stabilized pre-F trimer could elicit high titer neutralization, >10-fold higher than elicited by post-F. Immunogenicity assessments in pre-exposed rhesus macaques showed the triple disulfide-stabilized pre-F could recall high neutralizing titers after a single immunization, with little discrimination in the recall response between pre-F and post-F immunogens. However, the triple disulfide-stabilized pre-F adsorbed HMPV-directed responses from commercially available pooled human immunoglobulin more fully than post-F. Collectively, these results suggest single-chain triple disulfide-stabilized pre-F trimers to be promising HMPV-vaccine antigens.https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1011584&type=printable |
| spellingShingle | Li Ou Steven J Chen I-Ting Teng Lijuan Yang Baoshan Zhang Tongqing Zhou Andrea Biju Cheng Cheng Wing-Pui Kong Nicholas C Morano Erik-Stephane D Stancofski John-Paul Todd Yaroslav Tsybovsky Shuishu Wang Cheng-Yan Zheng John R Mascola Lawrence Shapiro Ruth A Woodward Ursula J Buchholz Peter D Kwong Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus. PLoS Pathogens |
| title | Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus. |
| title_full | Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus. |
| title_fullStr | Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus. |
| title_full_unstemmed | Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus. |
| title_short | Structure-based design of a single-chain triple-disulfide-stabilized fusion-glycoprotein trimer that elicits high-titer neutralizing responses against human metapneumovirus. |
| title_sort | structure based design of a single chain triple disulfide stabilized fusion glycoprotein trimer that elicits high titer neutralizing responses against human metapneumovirus |
| url | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1011584&type=printable |
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