Purpose <p>Respiratory syncytial virus (RSV) is a major pathogen affecting newborns and young toddlers, leading to approximately 30 million infections and 3 million hospitalizations annually. Due to limited treatment options, new prophylactic vaccines, such as ArexvyTM and AbrysvoTM, have been approved with some limitations. Similar to humoral immune response, CD8+T cells play a crucial role in controlling RSV infection. To activate CD8+T cells, this study focused on predicting potent CD8 + T cell epitopes against RSV using a combined in silico and in vivo approach.</p> Methods <p>This study utilized sequence- and structure-based methods to predict potent CD8 T-cell epitopes from the RSV proteome. IEDB-AR, NetCTLpan1.1, NetMHCcons1.1, NetMHC4, and NetMHC Pan4.1 were used to identify plausible CTL epitopes. Molecular docking analysis was conducted to characterize the molecular interactions of the predicted peptides with the murine H2-Kd allele. Subsequently, the immunological efficacy of the predicted CTL epitopes was evaluated in a BALB/c mouse model.</p> Results <p>In silico approaches revealed 21 CTL epitopes restricted by the murine H2-Kd allele, which were derived from the entire RSV proteome. Molecular docking studies revealed that 10 of the 21 epitopes showed favorable binding interactions with H2-Kd. Of these 10 epitopes, four epitopes were selected based on their affinity interactions for further experimental validation. The preclinical mice study demonstrated a significant elevation of IFN-γ and IL-2 cytokine responses, implying a potent CTL response to these four epitopes.</p> Conclusions <p>In conclusion, the application of a hybrid approach for the prediction of immunogenic epitopes offers valuable insights for rational vaccine development.</p> Graphical Abstract <p>Graphical representation of designing of vaccine model for respiratory syncytial virus</p> <p></p>