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Updated: Apr 4, 2026
Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses
Published on: April 4, 2019
A highly stable prefusion RSV F vaccine derived from structural analysis of the fusion mechanism
Anders Krarup1, Daphné Truan1, Polina Furmanova-Hollenstein1
1Janssen Infectious Diseases and Vaccines, Archimedesweg 4-6, Leiden 2333 CN, The Netherlands.
Insights
New strategies stabilize the respiratory syncytial virus (RSV) fusion protein (F) in its prefusion conformation. This stabilized RSV F antigen elicits neutralizing antibodies and provides complete protection against viral challenge in animal models.
Area of Science:
- Virology
- Immunology
- Structural Biology
Background:
- Respiratory syncytial virus (RSV) is a major cause of infant hospitalizations due to acute lower respiratory tract infections.
- The RSV fusion (F) protein is a key target for vaccines, but stabilizing its prefusion conformation is challenging.
Purpose of the Study:
- To develop alternative strategies for stabilizing the RSV F protein in its prefusion conformation.
- To identify mutations that enhance the stability and expression of the prefusion RSV F antigen.
Main Methods:
- Mutagenesis of the RSV F protein to prevent hinge movements and eliminate proteolytic exposure.
- Expression and purification of stabilized prefusion RSV F variants.
- Immunization of cotton rats and assessment of neutralizing antibody responses and protection against viral challenge.
- Structural and biochemical analyses of the stabilized variants.
Main Results:
- Identified unique mutations that effectively stabilize the prefusion conformation of RSV F.
- Achieved dramatically increased expression levels of the stabilized prefusion RSV F.
- Demonstrated that the stabilized prefusion RSV F elicits potent neutralizing antibodies in cotton rats.
- Showed complete protection against RSV viral challenge in immunized cotton rats.
- Structural analysis provided insights into the function of the p27 segment.
Conclusions:
- The developed strategies yield highly stable prefusion RSV F antigens with improved immunogenicity.
- These stabilized antigens represent promising candidates for effective RSV vaccines.
- Further understanding of RSV F structure and function was gained through the analysis of p27.
Abstract:
Respiratory syncytial virus (RSV) causes acute lower respiratory tract infections and is the leading cause of infant hospitalizations. Recently, a promising vaccine antigen based on the RSV fusion protein (RSV F) stabilized in the native prefusion conformation has been described. Here we report alternative strategies to arrest RSV F in the prefusion conformation based on the prevention of hinge movements in the first refolding region and the elimination of proteolytic exposure of the fusion peptide. A limited number of unique mutations are identified that stabilize the prefusion conformation of RSV F and dramatically increase expression levels. This highly stable prefusion RSV F elicits neutralizing antibodies in cotton rats and induces complete protection against viral challenge. Moreover, the structural and biochemical analysis of the prefusion variants suggests a function for p27, the excised segment that precedes the fusion peptide in the polypeptide chain.
