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Previously, we showed that inactivation of either the manganese- and iron-binding (EfaA) or zinc-binding (AdcA and AdcAII) lipoproteins significantly reduced E. faecalis virulence. Here, we explored whether immunization using a multi-valent approach induces protective immunity against systemic enterococcal infections. We found that multi-antigen antisera raised against EfaA, AdcA, and AdcAII displayed similar capacities to initiate neutrophil-mediated opsonization, like their single-antigen counterparts. Further, these antigen-specific antibodies worked synergistically with calprotectin, a divalent host metal chelator, to inhibit the growth of E. faecalis in laboratory media as well as in human sera. Using the Galleria mellonella invertebrate model and mouse peritonitis model, we showed that passive immunization with multi-antigen antisera conferred robust protection against E. faecalis infection, while the protective effects of single antigen antisera were negligible in G. mellonella, and negligible-to-moderate in the mouse model. Lastly, active immunization with the 3-antigen (trivalent) cocktail significantly protected mice against either lethal or non-lethal E. faecalis infections, with this protection appearing to be far-reaching based on immunization results obtained with contemporary strains of E. faecalis and closely related Enterococcus faecium.<\/jats:p>","DOI":"10.1093\/femsmc\/xtae031","type":"journal-article","created":{"date-parts":[[2024,10,4]],"date-time":"2024-10-04T03:51:00Z","timestamp":1728013860000},"source":"Crossref","is-referenced-by-count":6,"title":["Trivalent immunization with metal-binding proteins confers protection against enterococci in a mouse infection model"],"prefix":"10.1093","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0568-8628","authenticated-orcid":false,"given":"Ling Ning","family":"Lam","sequence":"first","affiliation":[{"name":"Department of Oral Biology, College of Dentistry, University of Florida , Gainesville, FL 32610 ,","place":["United 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