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Surface display of functional fibronectin-binding domains on Staphylococcus carnosus
S Liljeqvist1, F Cano, T N Nguyen
1Department of Biotechnology, Royal Institute of Technology (KTH), Stockholm, Sweden.
FEBS Letters
|April 1, 1999
Summary
Researchers engineered Staphylococcus carnosus to display fibronectin binding domains (FNBDs) on its surface. These engineered bacteria successfully bound fibronectin and enhanced immune responses, showing potential for live bacterial vaccine delivery systems.
Area of Science:
- Microbiology and Immunology
- Bacterial Surface Display
- Vaccine Development
Background:
- Fibronectin binding proteins (FNBPs) are crucial virulence factors for bacteria like Streptococcus dysgalactiae and Staphylococcus aureus.
- Engineering bacterial surfaces to display specific protein domains offers a platform for novel biotechnological applications.
- Developing effective live bacterial vaccine delivery systems requires robust methods for antigen display and immunogenicity.
Purpose of the Study:
- To investigate the surface expression and fibronectin-binding capabilities of different fibronectin binding domains (FNBDs) in Staphylococcus carnosus.
- To evaluate the potential of surface-displayed FNBDs in recombinant bacteria as a component of a live bacterial vaccine delivery system.
- To assess the immunogenicity of a model antigen presented on the surface of engineered bacteria.
Main Methods:
- Construction and surface expression of chimeric proteins containing FNBDs from S. dysgalactiae and S. aureus in Staphylococcus carnosus.
- Demonstration of surface localization of the chimeric proteins using appropriate assays.
- Whole-cell enzyme-linked binding assays to confirm fibronectin binding by surface-displayed FNBDs.
- Intranasal immunization studies in a model system to evaluate antibody responses to an antigen co-displayed on the bacterial surface.
Main Results:
- Successful surface expression and localization of three different FNBDs in Staphylococcus carnosus were confirmed.
- All surface-displayed FNBDs demonstrated significant binding to fibronectin.
- Intranasal immunization with one of the recombinant bacterial constructs led to enhanced antibody responses against a model immunogen.
Conclusions:
- Staphylococcus carnosus can be effectively engineered for surface display of functional fibronectin binding domains.
- Surface-displayed FNBDs retain their fibronectin-binding activity and can contribute to the immunogenicity of co-displayed antigens.
- These findings support the potential of using engineered bacteria displaying specific protein domains as platforms for live bacterial vaccine delivery.