Immunodominance of Adenovirus-Derived CD8 ⁺ T Cell Epitopes Interferes with the Induction of Transgene-Specific Immunity in Adenovirus-Based Immunization

Abstract

Adenovirus (Ad)-based immunization is a popular approach in vaccine development, and Ad-based vectors are renowned for their potential to induce strong CD8⁺ T cell responses to the encoded transgene. Surprisingly, we previously found in the mouse Friend retrovirus (FV) model that Ad-based immunization did not induce CD8⁺ T cell responses to the FV Leader-Gag-derived immunodominant epitope GagL_85-93 We show now that induction of GagL_85-93-specific CD8⁺ T cells was highly effective when leader-Gag was delivered by plasmid DNA immunization, implying a role for Ad-derived epitopes in mediating unresponsiveness. By immunizing with DNA constructs encoding strings of GagL_85-93 and the two Ad-derived epitopes DNA-binding protein_418-426 (DBP_418-426) and hexon_486-494, we confirmed that Ad epitopes prevent induction of GagL_85-93-specific CD8⁺ T cells. Interestingly, while DBP_418-426 did not interfere with GagL_85-93-specific CD8⁺ T cell induction, the H-2D^d-restricted hexon_486-494 suppressed the CD8⁺ T cell response to the H-2D^b-restricted GagL_85-93 strongly in H-2^b/d mice but not in H-2^b/b mice. This finding indicates that competition occurs at the level of responding CD8⁺ T cells, and we could indeed demonstrate that coimmunization with an interleukin 2 (IL-2)-encoding plasmid restored GagL_85-93-specific CD8⁺ T cell responses to epitope strings in the presence of hexon_486-494 IL-2 codelivery did not restore GagL_85-93 responsiveness in Ad-based immunization, however, likely due to the presence of further epitopes in the Ad vector. Our findings show that seemingly immunodominant transgene epitopes can be dominated by Ad-derived epitopes. These findings underline the importance of thorough characterization of vaccine vectors, and modifications of vectors or immunogens may be required to prevent impaired transgene-specific immune responses.<b>IMPORTANCE</b> Ad-based vectors are widely used in experimental preclinical and clinical immunization studies against numerous infectious agents, such as human immunodeficiency virus, Ebola virus, <i>Plasmodium falciparum</i>, or <i>Mycobacterium tuberculosis</i> Preexisting immunity to Ad-based vectors is widely recognized as a hindrance to the widespread use of Ad-based vectors for immunizations in humans; however, our data show that an immune response to Ad-derived T cell epitopes can also result in loss or impairment of transgene-specific immune responses in prenaive vaccinees due to immune competition. Our results highlight that seemingly immunodominant epitopes may be affected by dominance of vector-derived epitopes, and modifications of the vector design or the immunogens employed in immunization may lead to more effective vaccines.