Membrane association of a model CD4⁺ T-cell vaccine antigen confers enhanced yet incomplete protection against murid herpesvirus-4 infection

Vaccination against γ-herpesviruses has proved difficult. CD4⁺ T cells are essential to contain infection, but how best to prime them and whether this can reduce viral loads remain unclear. To address these questions, we used ovalbumin (OVA) as a model antigen, delivering it with murine cytomegalovirus (MCMV) to protect mice against OVA-expressing murine herpesvirus-4 (MuHV-4). Membrane-associated OVA (mOVA) was more effective than soluble OVA, both to prime CD4⁺ T cells and as an effector target. It was also a better target than an OVA epitope limited to infected cells, suggesting that protective CD4⁺ T cells recognize infected cell debris rather than infected cells themselves. While MCMV-mOVA protected acutely against MuHV-4-mOVA, long-term protection was incomplete, even when OVA-specific CD8⁺ T cells and B cells were also primed. Thus, even optimized single-target vaccines may poorly reduce long-term γ-herpesvirus infections.