Recombinant respiratory syncytial virus (RSV) A2-line19F strain with a point mutation and a protein encoding plasmid for expressing protein with same mutation for use in vaccine research for RSV.
- Mutated fusion protein shows a greater than 6-fold increase in fusion activity over line19 F protein.
- Mutated recombinant virus shows greater thermostabitily compared to parent line.
RSV is a ubiquitous virus worldwide and is the most common cause of bronchiolitis in children under the age of 1. The CDC estimates that almost all children will have had an RSV infection by age 2, with 2% requiring hospitalization. Severe infection rates are also increasing in the elderly population. Currently, treatment options for severe cases are limited, and a vaccine has yet to be developed. Research on RSV is impeded by its poor growth in tissue culture, lengthy replication cycle, and virion instability. Further development of research tools for RSV would increase the potential for vaccine development.
A recombinant respiratory syncytial virus (RSV) was generated from the A2-line 19F strain. This recombinant virus contains a point mutation at position F357 where the lysine residue was changed to a threonine (virus name- A2-line 19F-K357T). A protein expression plasmid encoding for the line19F protein with the same point mutation was also generated (protein name- line19F-K357T). The mutated protein has been shown to have higher fusion activity than the native line19 F protein, and the mutated virus has increase thermostablitiy, an important consideration for vaccine design and development.
Mutated virus and plasmid have been generated and preliminary fusion activity tested using dual split protein fusion assay in 293T cells.