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Virus Res. 2015 Dec 2;210:149-53. doi: 10.1016/j.virusres.2015.08.005. Epub 2015 Aug 10.

Identification of CD8 T cell epitopes in VP2 and NS1 proteins of African horse sickness virus in IFNAR(-/-) mice.

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Centro de Investigación en Sanidad Animal, INIA-CISA, Valdeolmos, 28130 Madrid, Spain.
The Pirbright Institute, Ash Road, Pirbright, Surrey GU24 0NF, UK.
Centro de Investigación en Sanidad Animal, INIA-CISA, Valdeolmos, 28130 Madrid, Spain. Electronic address:


African horse sickness virus (AHSV) is an Orbivirus of the family Reoviridae that causes severe pathology in equids. Previous work in our laboratory showed the presence of AHSV-specific CD8(+) T cells in mice immunized with recombinant Modified Vaccinia Ankara (rMVA) expressing VP2 and NS1 proteins. In the present work, we selected potential CD8 T cell epitopes (MHC-class I binding peptides) for the 129 mouse strain from the VP2 and NS1 proteins of AHSV-4, using a combination of four epitope prediction algorithms (SYFPEITHI, BYMAS, NetMHC I and NetMHCpan). ELISPOT and Intracellular Cytokine Staining (ICS) analysis showed that the VP2-720 (MSLLNFGAV), VP2-1044 (YTFGNKFLL), and NS1-83 (CVIKNADYV) peptides elicited IFN-γ production in splenocytes of MVA-VP2 and MVA-NS1 immunized mice and were identified as CD8(+) T cell epitopes. In addition, these three MHC-class I-binding peptides induced the expression of CD107a in CD8(+) T cells, an indirect marker of cytotoxic activity. Importantly, VP2-1044 and NS1-83 epitopes are conserved among all nine AHSV serotypes. These data demonstrate the activation of AHSV specific T-cell epitopes during vaccination with rMVAs expressing VP2 and NS1. Furthermore, the characterization of these CD8(+) T-cell epitopes provides information useful for the design of novel marker multiserotype vaccines against AHSV.


African horse sickness virus; CD8(+) T cell epitopes; IFNAR(-/-) mice; MVA-NS1; MVA-VP2

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