Resumen: Biofilm formation is a general strategy for bacterial pathogens to withstand host defense mechanisms. In this study, we found that serum proteases inhibit biofilm formation by Neisseria meningitidis, Neisseria gonorrhoeae, Haemophilus influenzae, and Bordetella pertussis. Confocal laser-scanning microscopy analysis revealed that these proteins reduce the biomass and alter the architecture of meningococcal biofilms. To understand the underlying mechanism, the serum was fractionated through size-exclusion chromatography and anion-exchange chromatography, and the composition of the fractions that retained anti-biofilm activity against N. meningitidis was analyzed by intensity-based absolute quantification mass spectrometry. Among the identified serum proteins, plasma kallikrein (PKLK), FXIIa, and plasmin were found to cleave neisserial heparin-binding antigen and the alpha-peptide of IgA protease on the meningococcal cell surface, resulting in the release of positively charged polypeptides implicated in biofilm formation by binding extracellular DNA. Further experiments also revealed that plasmin and PKLK inhibited biofilm formation of B. pertussis by cleaving filamentous hemagglutinin. We conclude that the proteolytic activity of serum proteases toward bacterial adhesins involved in biofilm formation could constitute a defense mechanism for the clearance of pathogens. Idioma: Inglés DOI: 10.1080/21505594.2021.2003115 Año: 2021 Publicado en: Virulence 12, 1 (2021), 2902-2917 ISSN: 2150-5594 Factor impacto JCR: 5.428 (2021) Categ. JCR: IMMUNOLOGY rank: 65 / 163 = 0.399 (2021) - Q2 - T2 Categ. JCR: MICROBIOLOGY rank: 43 / 138 = 0.312 (2021) - Q2 - T1 Categ. JCR: INFECTIOUS DISEASES rank: 36 / 96 = 0.375 (2021) - Q2 - T2 Factor impacto CITESCORE: 7.1 - Medicine (Q1) - Immunology and Microbiology (Q2)