Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model
ABSTRACT The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the prime target for vaccine development. The spike protein mediates both binding to host cells and membrane fusion and is also so far the only known viral target of neutralizing antibod...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2021-04-01
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| Series: | mBio |
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| Online Access: | https://journals.asm.org/doi/10.1128/mBio.02648-20 |
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| author | Fatima Amanat Shirin Strohmeier Raveen Rathnasinghe Michael Schotsaert Lynda Coughlan Adolfo García-Sastre Florian Krammer |
| author_facet | Fatima Amanat Shirin Strohmeier Raveen Rathnasinghe Michael Schotsaert Lynda Coughlan Adolfo García-Sastre Florian Krammer |
| author_sort | Fatima Amanat |
| collection | DOAJ |
| description | ABSTRACT The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the prime target for vaccine development. The spike protein mediates both binding to host cells and membrane fusion and is also so far the only known viral target of neutralizing antibodies. Coronavirus spike proteins are large trimers that are relatively unstable, a feature that might be enhanced by the presence of a polybasic cleavage site in SARS-CoV-2 spike. Exchange of K986 and V987 for prolines has been shown to stabilize the trimers of SARS-CoV-1 and the Middle East respiratory syndrome coronavirus spike proteins. Here, we test multiple versions of a soluble spike protein for their immunogenicity and protective effect against SARS-CoV-2 challenge in a mouse model that transiently expresses human angiotensin-converting enzyme 2 via adenovirus transduction. Variants tested include spike proteins with a deleted polybasic cleavage site, proline mutations, or a combination thereof, besides the wild-type protein. While all versions of the protein were able to induce neutralizing antibodies, only the antigen with both a deleted cleavage site and the K986P and V987P (PP) mutations completely protected from challenge in this mouse model. IMPORTANCE A vaccine for SARS-CoV-2 is urgently needed. A better understanding of antigen design and attributes that vaccine candidates need to have to induce protective immunity is of high importance. The data presented here validate the choice of antigens that contain the PP mutations and suggest that deletion of the polybasic cleavage site may lead to a further-optimized design. |
| first_indexed | 2024-12-14T06:26:53Z |
| format | Article |
| id | doaj.art-3115dd1525384801b253562bd43d4a68 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-14T06:26:53Z |
| publishDate | 2021-04-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-3115dd1525384801b253562bd43d4a682022-12-21T23:13:39ZengAmerican Society for MicrobiologymBio2150-75112021-04-0112210.1128/mBio.02648-20Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse ModelFatima Amanat0Shirin Strohmeier1Raveen Rathnasinghe2Michael Schotsaert3Lynda Coughlan4Adolfo García-Sastre5Florian Krammer6Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USAABSTRACT The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the prime target for vaccine development. The spike protein mediates both binding to host cells and membrane fusion and is also so far the only known viral target of neutralizing antibodies. Coronavirus spike proteins are large trimers that are relatively unstable, a feature that might be enhanced by the presence of a polybasic cleavage site in SARS-CoV-2 spike. Exchange of K986 and V987 for prolines has been shown to stabilize the trimers of SARS-CoV-1 and the Middle East respiratory syndrome coronavirus spike proteins. Here, we test multiple versions of a soluble spike protein for their immunogenicity and protective effect against SARS-CoV-2 challenge in a mouse model that transiently expresses human angiotensin-converting enzyme 2 via adenovirus transduction. Variants tested include spike proteins with a deleted polybasic cleavage site, proline mutations, or a combination thereof, besides the wild-type protein. While all versions of the protein were able to induce neutralizing antibodies, only the antigen with both a deleted cleavage site and the K986P and V987P (PP) mutations completely protected from challenge in this mouse model. IMPORTANCE A vaccine for SARS-CoV-2 is urgently needed. A better understanding of antigen design and attributes that vaccine candidates need to have to induce protective immunity is of high importance. The data presented here validate the choice of antigens that contain the PP mutations and suggest that deletion of the polybasic cleavage site may lead to a further-optimized design.https://journals.asm.org/doi/10.1128/mBio.02648-20COVID-19SARS-CoV-2spikevaccine |
| spellingShingle | Fatima Amanat Shirin Strohmeier Raveen Rathnasinghe Michael Schotsaert Lynda Coughlan Adolfo García-Sastre Florian Krammer Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model mBio COVID-19 SARS-CoV-2 spike vaccine |
| title | Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model |
| title_full | Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model |
| title_fullStr | Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model |
| title_full_unstemmed | Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model |
| title_short | Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model |
| title_sort | introduction of two prolines and removal of the polybasic cleavage site lead to higher efficacy of a recombinant spike based sars cov 2 vaccine in the mouse model |
| topic | COVID-19 SARS-CoV-2 spike vaccine |
| url | https://journals.asm.org/doi/10.1128/mBio.02648-20 |
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