An mRNA-Lipid Nanoparticle Platform Encoding the Conserved Outer Membrane Protein BamA Elicits Broadly Cross-Reactive Systemic and Mucosal Antibodies Against Antimicrobial-Resistant Neisseria gonorrhoeae

Abstract

Neisseria gonorrhoeae (Ng) is the causative agent of gonorrhea, and the global spread of antimicrobial-resistant strains makes vaccine development a public health priority. Although messenger RNA (mRNA) vaccines have transformed protection against viral diseases, the platform remains in its infancy against pathogenic bacteria. Here, we evaluated the immunogenicity and protective efficacy of an mRNA-lipid nanoparticle (LNP) vaccine encoding the highly conserved outer membrane antigen BamA in the female mouse model of lower genital tract infection. We delivered BamA mRNA-LNPs via intramuscular (IM) or intranasal (IN) routes, with or without CpG ODN 2395, and measured antigen-specific antibody responses in serum and vaginal lavage samples. Both routes elicited robust BamA-specific antibodies that recognized diverse Ng isolates, including ceftriaxone-resistant strains. However, neither route accelerated bacterial clearance nor reduced bioburden, nor did either generate serum bactericidal activity. These findings show that BamA mRNA-LNPs are immunogenic but, as formulated, are not protective, and they pave the way for modifications to the construct, adjuvant, and route. To our knowledge, this is the first evaluation of an mRNA vaccine against Ng, establishing the platform as an amenable approach for gonococcal antigen testing. ImportanceNeisseria gonorrhoeae is a high-priority public health threat, and no licensed gonococcal vaccine exists. mRNA-lipid nanoparticle vaccines have transformed antiviral immunization but remain largely untested against bacterial pathogens. This study is the first to evaluate an mRNA vaccine against N. gonorrhoeae, using the conserved, essential outer membrane antigen BamA. We show that BamA mRNA-LNPs delivered by intramuscular or intranasal routes elicit robust, broadly cross-reactive antibodies that recognize diverse isolates, including ceftriaxone-resistant strains, even though the tested formulations did not confer protection in mice. By establishing mRNA-LNPs as a viable platform for gonococcal antigen testing, this work lays the foundation for optimizing constructs, adjuvants, and routes for developing mRNA-based vaccines against gonorrhea and other antimicrobial-resistant bacteria.