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Bluetongue virus (BTV), an economically important orbivirus of the Reoviridae family, is a non-enveloped, dsRNA virus that causes a haemorrhagic disease mainly in sheep, but little is known of the cellular immunity elicited against BTV. We observed that vaccination of interferon type I-deficient mice (IFNAR((-/-))), or inoculation of the wild type C57BL/6 strain with BTV-8, induced a strong T cell response. Therefore, we proceeded to identify some of the T cell epitopes targeted by the immune system. We selected, using H-2(b)-binding predictive algorithms, 3 major histocompatibility complex (MHC)-class II-binding peptides and 7 MHC-class I binding peptides from the BTV-8 core protein VP7, as potential T cell epitopes. Peptide binding assays confirmed that all 7 MHC-class I predicted peptides bound MHC-class I molecules. Three MHC-class I and 2 MHC-class II binding peptide consistently elicited peptide-specific IFN-γ production (as measured by ELISPOT assays) in splenocytes from C57BL/6 BTV-8-inoculated mice and IFNAR((-/-))-vaccinated mice. The functionality of these T cells was confirmed by proliferation and cytotoxicity assays. Flow cytometry analysis demonstrated that CD8(+) T cells responded to MHC-class I binding peptides and CD4(+) T cells to MHC-class II binding peptides. Importantly, these 5 epitopes were also able to induced IFN-γ production in sheep inoculated with BTV-8. Taken together, these data demonstrate the activation of BTV-specific T cells during infection and vaccination. The characterisation of these novel T cell epitopes may also provide an opportunity to develop DIVA-compliant vaccination approach to BTV encompassing a broad-spectrum of serotypes.
CD4-Positive T-Lymphocytes, Immunity, Cellular, Sheep, Viral Core Proteins, Histocompatibility Antigens Class I, Histocompatibility Antigens Class II, Epitopes, T-Lymphocyte, CD8-Positive T-Lymphocytes, Lymphocyte Activation, Mice, Inbred C57BL, Interferon-gamma, Mice, Animals, Female, Peptides, Algorithms, Bluetongue virus, Protein Binding
CD4-Positive T-Lymphocytes, Immunity, Cellular, Sheep, Viral Core Proteins, Histocompatibility Antigens Class I, Histocompatibility Antigens Class II, Epitopes, T-Lymphocyte, CD8-Positive T-Lymphocytes, Lymphocyte Activation, Mice, Inbred C57BL, Interferon-gamma, Mice, Animals, Female, Peptides, Algorithms, Bluetongue virus, Protein Binding
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