Multimeric, Multivalent Fusion Carrier Proteins for Site-Selective Glycoconjugate Vaccines Simultaneously Targeting Staphylococcus aureus and Pseudomonas aeruginosa

Abstract

Staphylococcus aureus and Pseudomonas aeruginosa are major antimicrobial-resistant pathogens that often synergize in polymicrobial infections, such as chronic wound infections. These notorious and increasingly resistant bacteria contribute significantly to reduced antibiotic efficacy. Despite their substantial clinical burden, the urgent need to combat bacterial resistance and extensive research efforts, no vaccines currently exist for either bacterium. Glycoconjugate vaccines, which extend the range of suitable vaccine antigens to bacterial carbohydrates, could play a major role in this emergence. This study introduces a multiepitope vaccine conjugating S. aureus capsular polysaccharide serotype 8 to a chimeric protein fusing Hla and PcrV, two potent cytotoxins from S. aureus and P. aeruginosa, respectively. A conjugation strategy based on selective targeting of a purposefully introduced histidine tag was developed to preserve the structure and antigenicity of epitopes from the two proteins, leveraging their dual role as carrier and antigen. This multivalent, multimeric and multipathogen construct successfully elicited antibodies against all three antigens as well as functional protection. This proof-of-concept highlights the potential for advanced vaccines targeting polymicrobial infections and bacteria with complex pathogenesis calling for multivalent formulations. It also points out the power of site-selective conjugation as tool for vaccine manufacturing.