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A gel-free proteomic-based method for the characterization of Bordetella pertussis clinical isolates
Yulanda M Williamson1, Hercules Moura, Kaneatra Simmons
1Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Chamblee, Georgia 30341, USA.
Insights
Researchers analyzed surface proteins of Bordetella pertussis (Bp) to understand whooping cough. Proteomics identified changes in surface proteins, aiding in the development of improved vaccines and control strategies against this contagious respiratory disease.
Area of Science:
- Microbiology
- Immunology
- Proteomics
Background:
- Bordetella pertussis (Bp) causes pertussis (whooping cough), a significant public health concern, particularly in unvaccinated infants and young children.
- Increasing pertussis cases necessitate re-evaluation of current vaccine strategies and understanding of bacterial virulence factors.
- Bacterial outer membrane proteins (surfaceome) are crucial for pathogenesis and host immune responses, making them key targets for vaccine development.
Purpose of the Study:
- To identify and characterize changes in the surface protein expression (surfaceome) of recent Bordetella pertussis isolates.
- To assess the immunogenic potential of outer membrane proteins from different Bp strains using novel antibody capture technology.
- To provide insights into Bp pathogenesis and inform the development of next-generation pertussis vaccines.
Main Methods:
- Proteomic analysis using nano liquid chromatography-electrospray ionization-mass spectrometry (nLC-ESI-MS) to identify surface proteins in three Bp isolates.
- Enrichment of outer membrane protein fractions followed by trypsin digestion and peptide analysis.
- Application of a non-gel based antibody affinity capture technique coupled with MS to identify immunogenic surface proteins.
Main Results:
- Detailed characterization of the surfaceomes of three distinct Bordetella pertussis isolates was achieved.
- Identification of specific outer membrane proteins that are immunogenic and potentially involved in Bp pathogenesis.
- Demonstration of a novel, efficient method for identifying immunogenic bacterial surface proteins.
Conclusions:
- The study elucidates the surface protein landscape of circulating Bp strains, revealing potential targets for improved pertussis vaccines.
- The developed antibody affinity capture MS technique offers a broadly applicable tool for identifying immunogenic surface proteins in pathogenic bacteria.
- Findings contribute to a better understanding of Bp-host interactions and support the ongoing efforts to control pertussis outbreaks.
Abstract:
Bordetella pertussis (Bp) is the etiologic agent of pertussis or whooping cough, a highly contagious respiratory disease occurring primarily in infants and young children. Although vaccine preventable, pertussis cases have increased over the years leading researchers to re-evaluate vaccine control strategies. Since bacterial outer membrane proteins, comprising the surfaceome, often play roles in pathogenesis and antibody-mediated immunity, three recent Bp circulating isolates were examined using proteomics to identify any potential changes in surface protein expression. Fractions enriched for outer membrane proteins were digested with trypsin and the peptides analyzed by nano liquid chromatography-electrospray ionization-mass spectrometry (nLC-ESI-MS), followed by database analysis to elucidate the surfaceomes of our three Bp isolates. Furthermore, a less labor intensive non-gel based antibody affinity capture technology in conjunction with MS was employed to assess each Bp strains' immunogenic outer membrane proteins. This novel technique is generally applicable allowing for the identification of immunogenic surface expressed proteins on pertussis and other pathogenic bacteria.
