Based on the magnitude, virus-specific CD8⁺ cytotoxic T lymphocyte (CTL) responses directed against virus-derived peptide-MHC I complexes can be grouped as immunodominant and subdominant. Such immunodominance hierarchies are governed by multiple factors, including naïve T cell precursor frequency, proliferative capacity and clonotypic diversity of antigen (Ag)-specific CTLs. However, how these factors determine together inter/intra-individual immunodominance remains elusive. In this study, we dissected CTL immunodominance hierarchy for universally conserved influenza (IAV)-specific epitopes restricted by various HLAs by defining the magnitude of CTL memory populations relative to the HLA-A*02:01 responders. Our data from 67 donors covering the HLA Class I types presenting universally conserved IAV-specific peptides, HLA-A*02:01, A*03:01, B*27:05, B*57:01, B*18:01 and B*08:01, revealed that CTLs directed against HLA-A*0201:M1₅₈ were immunodominant. Conversely, in A*02:01⁺B*27:05⁺ heterozygous individuals, HLA-B*27:05-NP₃₈₃ elicited higher magnitude CTL responses_. Subsequently, we further dissected the basis of B*27:05 immundomination over A*02:01 by analyses of T cell receptor (TCRab) repertoire, TCR avidity and kinetics of killing and proliferation. Our data show that following in vitro peptide stimulation, B*27:05-NP₃₈₃ CTLs proliferate more rapidly acquiring higher levels of polyfunctionality than A*02:01-M1₅₈ specific CTLs. Furthermore, ex vivo single-cell paired CDR3ab analysis revealed that public A*02:01-M1₅₈ TCRs characterized by Vb19/Va27 profiles in A*02:01 individuals are significantly reduced/absent in A*02:01⁺B*27:05⁺ individuals. Clonal analysis of M1₅₈ dominant public TCRs suggested B*27:05-mediated removal of self-reactive TCRs and is under investigation. Overall, our study provides insights into the importance of human IAV-specific immunodominance at quantitative and qualitative levels to modulate antiviral T cell responses for vaccine development.