A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization Response

<b>Background/Objectives:</b> Human metapneumovirus (hMPV) is a leading cause of respiratory infections in the elderly, with high morbidity and mortality and with no vaccines or specific therapies available. The primary protective antigen of hMPV is the fusion protein, and its prefusion...

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Main Authors: Michael Kishko, Antonia Stuebler, Sukanya Sasmal, Yvonne Chan, Dean Huang, Christopher Reyes, Jasmine Lin, Owen Price, Ana Kume, Katie Zong, Christine Bricault, Judith Alamares-Sapuay, Linong Zhang
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Language:English
Published: MDPI AG 2025-05-01
Series:Vaccines
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Online Access:https://www.mdpi.com/2076-393X/13/5/523
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author Michael Kishko
Antonia Stuebler
Sukanya Sasmal
Yvonne Chan
Dean Huang
Christopher Reyes
Jasmine Lin
Owen Price
Ana Kume
Katie Zong
Christine Bricault
Judith Alamares-Sapuay
Linong Zhang
author_facet Michael Kishko
Antonia Stuebler
Sukanya Sasmal
Yvonne Chan
Dean Huang
Christopher Reyes
Jasmine Lin
Owen Price
Ana Kume
Katie Zong
Christine Bricault
Judith Alamares-Sapuay
Linong Zhang
author_sort Michael Kishko
collection DOAJ
description <b>Background/Objectives:</b> Human metapneumovirus (hMPV) is a leading cause of respiratory infections in the elderly, with high morbidity and mortality and with no vaccines or specific therapies available. The primary protective antigen of hMPV is the fusion protein, and its prefusion conformation (pre-F) is considered the most promising target for vaccine development. <b>Methods:</b> Utilizing computational design strategies focused on intraprotomer interface stabilization, we designed hMPV pre-F recombinant subunit vaccine candidates based on the most prevalent A2 subtype and characterized them in vitro and in vivo, benchmarking to the prototypical hMPV pre-F stabilized by an introduction of a proline at site 185. <b>Results:</b> The top candidate (N46V_T160F) yielded 14.4 mg/L with a melting temperature of 79.3 °C as compared to 5.7 mg/L and 70.4 °C for the benchmark. By employing monoclonal antibody binding to all six antigenic sites of hMPV pre-F, we confirmed this construct retained all pre-F specific antigenic sites and that the key sites Ø and V were stable at 4 °C for up to 6 months. When immunogenicity of N46V_T160F was evaluated in mice, it induced higher binding and neutralizing antibody titers than the benchmark, which stemmed in part from increased levels of site Ø and site II targeting Abs. Further, this A2 based construct induced cross-neutralizing Abs against all four hMPV subtypes. Lastly, our construct exhibited similar immunogenicity as the recently published next-generation hMPV pre-F constructs, DS-CavEs2 and v3B_Δ12_D454C-V458C. <b>Conclusions:</b> N46V_T160F is a promising hMPV vaccine candidate paving the way for further development and optimization.
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spelling doaj-art-4e91f53dcce74a7f8e876c6254076d192025-08-20T02:33:55ZengMDPI AGVaccines2076-393X2025-05-0113552310.3390/vaccines13050523A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization ResponseMichael Kishko0Antonia Stuebler1Sukanya Sasmal2Yvonne Chan3Dean Huang4Christopher Reyes5Jasmine Lin6Owen Price7Ana Kume8Katie Zong9Christine Bricault10Judith Alamares-Sapuay11Linong Zhang12Sanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USASanofi, 200 West Street, Waltham, MA 02451, USA<b>Background/Objectives:</b> Human metapneumovirus (hMPV) is a leading cause of respiratory infections in the elderly, with high morbidity and mortality and with no vaccines or specific therapies available. The primary protective antigen of hMPV is the fusion protein, and its prefusion conformation (pre-F) is considered the most promising target for vaccine development. <b>Methods:</b> Utilizing computational design strategies focused on intraprotomer interface stabilization, we designed hMPV pre-F recombinant subunit vaccine candidates based on the most prevalent A2 subtype and characterized them in vitro and in vivo, benchmarking to the prototypical hMPV pre-F stabilized by an introduction of a proline at site 185. <b>Results:</b> The top candidate (N46V_T160F) yielded 14.4 mg/L with a melting temperature of 79.3 °C as compared to 5.7 mg/L and 70.4 °C for the benchmark. By employing monoclonal antibody binding to all six antigenic sites of hMPV pre-F, we confirmed this construct retained all pre-F specific antigenic sites and that the key sites Ø and V were stable at 4 °C for up to 6 months. When immunogenicity of N46V_T160F was evaluated in mice, it induced higher binding and neutralizing antibody titers than the benchmark, which stemmed in part from increased levels of site Ø and site II targeting Abs. Further, this A2 based construct induced cross-neutralizing Abs against all four hMPV subtypes. Lastly, our construct exhibited similar immunogenicity as the recently published next-generation hMPV pre-F constructs, DS-CavEs2 and v3B_Δ12_D454C-V458C. <b>Conclusions:</b> N46V_T160F is a promising hMPV vaccine candidate paving the way for further development and optimization.https://www.mdpi.com/2076-393X/13/5/523computational designhMPVsubunit vaccineprefusion stabilization
spellingShingle Michael Kishko
Antonia Stuebler
Sukanya Sasmal
Yvonne Chan
Dean Huang
Christopher Reyes
Jasmine Lin
Owen Price
Ana Kume
Katie Zong
Christine Bricault
Judith Alamares-Sapuay
Linong Zhang
A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization Response
Vaccines
computational design
hMPV
subunit vaccine
prefusion stabilization
title A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization Response
title_full A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization Response
title_fullStr A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization Response
title_full_unstemmed A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization Response
title_short A Computationally Designed Prefusion Stabilized Human Metapneumovirus Fusion Protein Vaccine Antigen Elicited a Potent Neutralization Response
title_sort computationally designed prefusion stabilized human metapneumovirus fusion protein vaccine antigen elicited a potent neutralization response
topic computational design
hMPV
subunit vaccine
prefusion stabilization
url https://www.mdpi.com/2076-393X/13/5/523
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