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This phenomenon can be exemplified by COVID-19 vaccines. COVID-19 vaccines, including mRNA platforms, use the SARS-CoV-2 spike protein as an immunogen to induce the production of neutralizing antibodies. The spike protein binds the ACE2 (angiotensin-converting enzyme 2) receptor on human cells, mediating viral entry and infection. ACE2 is widely expressed across multiple tissues and is a key component of the renin\u2013angiotensin\u2013aldosterone system (RAAS) that acts as a homeostatic regulator of systemic and local blood flow, blood pressure, cardiac function, fluid balance and immunity. Some studies have proposed the interaction between the spike protein and ACE2 as a possible contributing factor to rare adverse effects observed following COVID-19 vaccination, including myocarditis, pericarditis, thrombosis, and reported alterations in blood pressure, though these mechanisms remain to be fully elucidated. Objectives: As a proof-of-concept approach in vaccine antigen development, we engineered SARS-CoV-2 spike mutants with impaired binding to the host receptor ACE2. Methods: By rational design, we produced and validated in vitro and in vivo spike point mutants that do not effectively bind ACE2. Results: The engineered spike mutants do not effectively bind the human entry receptor ACE2 while retaining the immunogenic properties equal to or better than the wild type spike and thus generate a protective response in animals when used as a vaccination agent. Conclusions: By establishing a straightforward molecular strategy for rational vaccine design, this work demonstrates the feasibility of limiting specific antigen\u2013host receptor interactions while maintaining immunogenicity. This approach may be applicable to future vaccination strategies where antigen interaction with host cells could potentially interfere with physiological pathways.<\/jats:p>","DOI":"10.3390\/vaccines14070568","type":"journal-article","created":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T00:55:11Z","timestamp":1782780911000},"page":"568","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rationally Modified SARS-CoV-2 Spike Protein Impairs ACE2 Binding While Preserving Immunogenicity in Mice"],"prefix":"10.3390","volume":"14","author":[{"given":"Elia","family":"Tamagnini","sequence":"first","affiliation":[{"name":"Institute for Research in Biomedicine, Universit\u00e0 della Svizzera italiana (USI), 6500 Bellinzona, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca","family":"Simonelli","sequence":"additional","affiliation":[{"name":"Institute for Research in Biomedicine, Universit\u00e0 della Svizzera italiana (USI), 6500 Bellinzona, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4279-7092","authenticated-orcid":false,"given":"Martin","family":"Palus","sequence":"additional","affiliation":[{"name":"Laboratory of Arbovirology, Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, CZ-37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"},{"name":"Laboratory of Emerging Viral Infections, Veterinary Research Institute, CZ-62100 Brno, Czech Republic"},{"name":"Department of Experimental Biology, Faculty of Science, Masaryk University, CZ-62500 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-2146-2237","authenticated-orcid":false,"given":"Tanja Rezzonico","family":"Jost","sequence":"additional","affiliation":[{"name":"Institute of Oncology Research, 6500 Bellinzona, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6369-5442","authenticated-orcid":false,"given":"Edoardo","family":"Lazzarini","sequence":"additional","affiliation":[{"name":"Institute for Translational Research (IRT), Faculty of Biomedical Sciences, Universit\u00e0 Svizzera Italiana (USI), Ente Ospedaliero Cantonale (EOC), 6500 Bellinzona, Switzerland"},{"name":"Cardiovascular Theranostics Group, Istituto Cardiocentro Ticino, Ente Ospedaliero Cantonale (EOC), 6900 Lugano, Switzerland"},{"name":"Euler Institute, Faculty of Biomedical Sciences, Universit\u00e0 della Svizzera Italiana (USI), 6900 Lugano, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5297-0234","authenticated-orcid":false,"given":"Davide","family":"Mangani","sequence":"additional","affiliation":[{"name":"Institute for Research in Biomedicine, Universit\u00e0 della Svizzera italiana (USI), 6500 Bellinzona, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0469-4604","authenticated-orcid":false,"given":"V\u00e1clav","family":"H\u00f6nig","sequence":"additional","affiliation":[{"name":"Laboratory of Arbovirology, Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, CZ-37005 \u010cesk\u00e9 Bud\u011bjovice, Czech Republic"},{"name":"Laboratory of Emerging Viral Infections, Veterinary Research Institute, CZ-62100 Brno, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-0276-9065","authenticated-orcid":false,"given":"Mark\u00e9ta","family":"Dvo\u0159\u00e1kov\u00e1","sequence":"additional","affiliation":[{"name":"Laboratory of Arbovirology, Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, CZ-37005 \u010cesk\u00e9 Bud\u011bjovice, Czech 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Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3348-1712","authenticated-orcid":false,"given":"Fabio","family":"Grassi","sequence":"additional","affiliation":[{"name":"National Institute for Molecular Genetics \u201cRomeo ed Enrica Invernizzi\u201d INGM, 20122 Milan, Italy"},{"name":"Department of Medical Biotechnology and Translational Medicine, University of Milan, 20129 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5827-0439","authenticated-orcid":false,"given":"Lucio","family":"Barile","sequence":"additional","affiliation":[{"name":"Institute for Translational Research (IRT), Faculty of Biomedical Sciences, Universit\u00e0 Svizzera Italiana (USI), Ente Ospedaliero Cantonale (EOC), 6500 Bellinzona, Switzerland"},{"name":"Cardiovascular Theranostics Group, Istituto Cardiocentro Ticino, Ente Ospedaliero Cantonale (EOC), 6900 Lugano, Switzerland"},{"name":"Euler 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