Identification of CD8 T cell epitopes in VP2 and NS1 proteins of African horse sickness virus in IFNAR(-/-) mice.
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.
Centro de Investigación en Sanidad Animal, INIA-CISA, Valdeolmos, 28130 Madrid, Spain.
Francisco
F
de la Poza
Centro de Investigación en Sanidad Animal, INIA-CISA, Valdeolmos, 28130 Madrid, Spain.
Alejandro
A
Marín-López
The Pirbright Institute, Ash Road, Pirbright, Surrey GU24 0NF, UK.
Javier
J
Castillo-Olivares
The Pirbright Institute, Ash Road, Pirbright, Surrey GU24 0NF, UK.
Eva
E
Calvo-Pinilla
Centro de Investigación en Sanidad Animal, INIA-CISA, Valdeolmos, 28130 Madrid, Spain. Electronic address: ortego@inia.es.
Javier
J
Ortego
D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D036022:Capsid Proteins / Q000276:immunology*; D019295:Computational Biology; D058501:Enzyme-Linked Immunospot Assay; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000032:analysis*; D007371:Interferon-gamma / Q000032:analysis; D051907:Lysosomal Membrane Proteins / Q000032:analysis; D051379:Mice; D018345:Mice, Knockout; D053633:Receptor, Interferon alpha-beta / Q000172:deficiency*; D013194:Staining and Labeling; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000008:administration & dosage; D052118:Lysosomal-Associated Membrane Protein 1
D036022:Capsid Proteins; D018984:Epitopes, T-Lymphocyte; D007371:Interferon-gamma; D051907:Lysosomal Membrane Proteins; D053633:Receptor, Interferon alpha-beta; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C052255:Lamp1 protein, mouse; D052118:Lysosomal-Associated Membrane Protein 1; C089209:NS1 protein, African horse sickness virus; C442519:VP2 protein, African horse sickness virus
African horse sickness virus; CD8(+) T cell epitopes; IFNAR(-/-) mice; MVA-NS1; MVA-VP2