Abstract
Bluetongue virus (BTV) is a non-enveloped dsRNA virus that causes a haemorrhagic disease mainly in sheep. It is an economically important Orbivirus of the Reoviridae family. In order to estimate the importance of T cell responses during BTV infection, it is essential to identify the epitopes targeted by the immune system. In the present work, we selected potential T cell epitopes (3 MHC-class II-binding and 8 MHC-class I binding peptides) for the C57BL/6 mouse strain from the BTV-8 non-structural protein NS1, using H2b-binding predictive algorithms. Peptide binding assays confirmed all MHC-class I predicted peptides bound MHC-class I molecules. The immunogenicity of these 11 predicted peptides was then determined using splenocytes from BTV-8-inoculated C57BL/6 mice. Four MHC-class I binding peptides elicited specific IFN-γ production and generated cytotoxic T lymphocytes (CTL) in BTV-8 infected mice. CTL specific for 2 of these peptides were also able to recognise target cells infected with different BTV serotypes. Similarly, using a combination of IFN-γ ELISPOT, intracellular cytokine staining and proliferation assays, two MHC-class II peptides were identified as CD4+ T cell epitopes in BTV-8 infected mice. Importantly, two peptides were also consistently immunogenic in sheep infected with BTV-8 using IFN-γ ELISPOT assays. Both of these peptides stimulated CD4+ T cells that cross-reacted with other BTV serotypes. The characterisation of these T cell epitopes can help develop vaccines protecting against a broad spectrum of BTV serotypes and differentiate infected from vaccinated animals.
Highlights
Bluetongue virus (BTV) is the prototype member of the Orbivirus genus of the Reoviridae family, transmitted to the vertebrate host by biting midges [1]
Infection with BTV-8 induces T cell responses to other BTV serotypes in mice and sheep Splenocytes from BTV-8 inoculated mice were cultured in the presence of inactivated virus
In the present work, we described cytotoxic T lymphocytes (CTL) and T helper epitopes from the non-structural protein 1 (NS1) protein of BTV-8 in mice and sheep
Summary
Bluetongue virus (BTV) is the prototype member of the Orbivirus genus of the Reoviridae family, transmitted to the vertebrate host by biting midges [1]. A long-lasting immunity is developed in animals that recover from bluetongue where both neutralising antibodies [10] and cytotoxic T lymphocytes (CTL) [11,12] are involved in this protective immunity. The variability of the outer capsid of this virus represents one of the major challenges for the development of a vaccine capable of protecting animals against multiple serotypes. BTV vaccination and infection in sheep induces CTLs cross-reactive to multiple serotypes [11,16,17,18]. Based on this observation, vaccination designed to elicit T cell responses can potentially protect animals against several BTV serotypes
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