{"doi":"10.1128/jvi.00087-25","title":"Hydrophobic residue substitutions enhance the stability and\n                    <i>in vivo</i>\n                    immunogenicity of respiratory syncytial virus fusion protein","abstract":"<jats:title>ABSTRACT</jats:title>\n                  <jats:sec>\n                    <jats:title/>\n                    <jats:p>Respiratory syncytial virus (RSV) entry into host cells is facilitated by viral fusion, wherein the metastable RSV fusion (F) protein undergoes a conformational change from a prefusion state to a highly stable postfusion structure. The prefusion F elicits a more robust human antibody response than its postfusion F and is a primary target for RSV vaccine development. However, the inherent instability of the prefusion F trimer and its low protein expression level in host cells are a significant challenge for developing a high-potency RSV vaccine. Here, we report that the introduction of four hydrophobic residue substitutions in the RSV F protein resulted in a highly stable prefusion F trimer (pre-F-IFLP). This engineered variant exhibits enhanced expression and stability compared to DS-Cav1, with improved thermal stability, increased resistance to acid and base, and extended storage life. Furthermore, pre-F-IFLP induced neutralizing antibody responses 72-fold higher than those elicited by DS-Cav1 following a second booster immunization and fully protected mice against RSV infection.</jats:p>\n                  </jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE</jats:title>\n                    <jats:p>\n                      In this study, we demonstrate that introducing four hydrophobic residue substitutions into the RSV F protein leads to the generation of a highly stable prefusion F trimer (pre-F-IFLP) with improved expression levels in cultured cells and superior stability compared to DS-Cav1, the first-generation prefusion F-stabilized RSV vaccine. Furthermore, pre-F-IFLP induced significantly higher neutralizing antibody responses than DS-Cav1 following both the first and second booster immunizations and conferred complete protection against RSV infection in a mouse model. These findings present an alternative approach for stabilizing the trimeric prefusion F protein, enhancing its expression, and significantly improving its protective efficacy for the prevention of RSV infection\n                      <jats:italic toggle=\"yes\">in vivo</jats:italic>\n                      .\n                    </jats:p>\n                  </jats:sec>","journal":"Journal of Virology","year":2025,"id":638667,"datarank":0.20794415416798362,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"self_citation_contribution":0.20794415416798362,"citation_network_contribution":0.0,"self_endowment_contribution":0.20794415416798362,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":848422,"name":"Haixia Yang","orcid":"0000-0002-4410-9257","position":1,"is_corresponding":false},{"id":1009804,"name":"Haoyue Zhu","orcid":"0009-0008-6609-398X","position":2,"is_corresponding":false},{"id":268453,"name":"Yun Hu","orcid":"0000-0001-5597-9234","position":3,"is_corresponding":false},{"id":1658995,"name":"Wenling Shen","orcid":null,"position":4,"is_corresponding":false},{"id":1658996,"name":"Huifeng Cheng","orcid":null,"position":5,"is_corresponding":false},{"id":1658997,"name":"Jialiao Cai","orcid":null,"position":6,"is_corresponding":false},{"id":1658998,"name":"Manlan Qiu","orcid":null,"position":7,"is_corresponding":false},{"id":1658999,"name":"Yueyue Li","orcid":null,"position":8,"is_corresponding":false},{"id":1659000,"name":"Yaolan Li","orcid":null,"position":9,"is_corresponding":false},{"id":1614619,"name":"Wencai Ye","orcid":"0000-0002-2810-1001","position":10,"is_corresponding":false},{"id":628578,"name":"Ying Wang","orcid":"0000-0003-4524-1812","position":11,"is_corresponding":false},{"id":603326,"name":"Wei Tang","orcid":"0000-0002-1990-4558","position":12,"is_corresponding":false},{"id":1658994,"name":"Qiaoyun Song","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Hydrophobic residue substitutions enhance the stability and\n                    <i>in vivo</i>\n                    immunogenicity of respiratory syncytial virus fusion protein","abstract":"<jats:title>ABSTRACT</jats:title>\n                  <jats:sec>\n                    <jats:title/>\n                    <jats:p>Respiratory syncytial virus (RSV) entry into host cells is facilitated by viral fusion, wherein the metastable RSV fusion (F) protein undergoes a conformational change from a prefusion state to a highly stable postfusion structure. The prefusion F elicits a more robust human antibody response than its postfusion F and is a primary target for RSV vaccine development. However, the inherent instability of the prefusion F trimer and its low protein expression level in host cells are a significant challenge for developing a high-potency RSV vaccine. Here, we report that the introduction of four hydrophobic residue substitutions in the RSV F protein resulted in a highly stable prefusion F trimer (pre-F-IFLP). This engineered variant exhibits enhanced expression and stability compared to DS-Cav1, with improved thermal stability, increased resistance to acid and base, and extended storage life. Furthermore, pre-F-IFLP induced neutralizing antibody responses 72-fold higher than those elicited by DS-Cav1 following a second booster immunization and fully protected mice against RSV infection.</jats:p>\n                  </jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE</jats:title>\n                    <jats:p>\n                      In this study, we demonstrate that introducing four hydrophobic residue substitutions into the RSV F protein leads to the generation of a highly stable prefusion F trimer (pre-F-IFLP) with improved expression levels in cultured cells and superior stability compared to DS-Cav1, the first-generation prefusion F-stabilized RSV vaccine. Furthermore, pre-F-IFLP induced significantly higher neutralizing antibody responses than DS-Cav1 following both the first and second booster immunizations and conferred complete protection against RSV infection in a mouse model. These findings present an alternative approach for stabilizing the trimeric prefusion F protein, enhancing its expression, and significantly improving its protective efficacy for the prevention of RSV infection\n                      <jats:italic toggle=\"yes\">in vivo</jats:italic>\n                      .\n                    </jats:p>\n                  </jats:sec>","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"40434102","pmcid":null,"openalex_id":null,"authors":[],"funders":[{"funder_name":"National Natural Science Foundation of China","grant_id":"82293681(82293680)","title":null},{"funder_name":"National Natural Science Foundation of China","grant_id":"82321004","title":null},{"funder_name":"National Natural Science Foundation of China","grant_id":"82003610","title":null},{"funder_name":"Guangdong Basic and Applied Basic Research Foundation","grant_id":"2022B1515120015, 2024B1515040014","title":null},{"funder_name":"Guangdong Major Project of Basic and Applied Basic Research","grant_id":"2023B0303000026","title":null},{"funder_name":"Natural Science Foundation of Guangdong Province","grant_id":"2025A1515011828, 2023A1515011113","title":null},{"funder_name":"Fundamental Research Funds for the Central Universities","grant_id":"21624224","title":null},{"funder_name":"Guangzhou Basic and Applied Basic Research Subject","grant_id":"2024A04J4171","title":null}],"total_grants":8,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://doi.org/10.1128/jvi.00087-25","host_type":"GOLD"},{"url":"https://doi.org/10.1128/jvi.00087-25","host_type":"publisher"},{"url":"https://journals.asm.org/doi/pdf/10.1128/jvi.00087-25","host_type":"publisher"},{"url":"https://doaj.org/article/c5140d3870a942b9882a9c38e67b2de7","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/12172430","host_type":"repository"}],"fields_of_study":["Medicine","Chemistry","Animals","Viral Fusion Proteins","Respiratory Syncytial Virus Infections","Mice","Antibodies, Viral","Antibodies, Neutralizing","Humans","Respiratory Syncytial Virus Vaccines","Respiratory Syncytial Virus, Human","Amino Acid Substitution","Mice, Inbred BALB C","Hydrophobic and Hydrophilic Interactions","Protein Stability","Female","Immunogenicity, Vaccine"],"mesh_terms":["Animals","Viral Fusion Proteins","Respiratory Syncytial Virus Infections","Mice","Antibodies, Viral","Antibodies, Neutralizing","Humans","Respiratory Syncytial Virus Vaccines","Respiratory Syncytial Virus, Human","Amino Acid Substitution","Mice, Inbred BALB C","Hydrophobic and Hydrophilic Interactions","Protein Stability","Female","Immunogenicity, Vaccine"],"keywords":["prefusion F","respiratory syncytial virus","vaccine","viral fusion"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-06T21:09:33.756029Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}