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Studies of immune responses elicited by bovine viral diarrhea virus (BVDV) vaccines have primarily focused on the characterization of neutralizing B cell and CD4 + T cell epitopes. Despite the availability of commercial vaccines for decades, BVDV prevalence in cattle has remained largely unaffected. There is limited knowledge regarding the role of BVDV-specific CD8 + T cells in immune protection, and indirect evidence suggests that they play a crucial role during BVDV infection. In this study, the presence of BVDV-specific CD8 + T cells that are highly cross-reactive in cattle was demonstrated. Most importantly, novel potent IFN-γ-inducing CD8 + T cell epitopes were identified from different regions of BVDV polyprotein. Eight CD8 + T cell epitopes were identified from the following structural BVDV Ags: E rns , E1, and E2 glycoproteins. In addition, from nonstructural BVDV Ags N pro , NS2-3, NS4A-B, and NS5A-B, 20 CD8 + T cell epitopes were identified. The majority of these IFN-γ-inducing CD8 + T cell epitopes were found to be highly conserved among more than 200 strains from BVDV-1 and -2 genotypes. These conserved epitopes were also validated as cross-reactive because they induced high recall IFN-γ + CD8 + T cell responses ex vivo in purified bovine CD8 + T cells isolated from BVDV-1- and -2-immunized cattle. Altogether, 28 bovine MHC class I-binding epitopes were identified from key BVDV Ags that can elicit broadly reactive CD8 + T cells against diverse BVDV strains. The data presented in this study will lay the groundwork for the development of a contemporary CD8 + T cell-based BVDV vaccine capable of addressing BVDV heterogeneity more effectively than current vaccines.

Novel Potent IFN-γ–Inducing CD8+ T Cell Epitopes Conserved among Diverse Bovine Viral Diarrhea Virus Strains