Use of “one-pot, mix-and-read” peptide-MHC class I tetramers and predictive algorithms to improve detection of cytotoxic T lymphocyte responses in cattle

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dc.creatorSvitek, Nicholas
dc.creatorHansen, A.M.
dc.creatorSteinaa, Lucilla
dc.creatorSaya, R.
dc.creatorAwino, Elias
dc.creatorNielsen, Morten
dc.creatorBuus, Soren
dc.creatorNene, Vishvanath M.
dc.date2014
dc.date2014-06-24T10:27:53Z
dc.date2014-06-24T10:27:53Z
dc.date.accessioned2026-06-27T17:18:39Z
dc.descriptionPeptide-major histocompatibility complex (p-MHC) class I tetramer complexes have facilitated the early detection and functional characterisation of epitope specific CD8+ cytotoxic T lymphocytes (CTL). Here, we report on the generation of seven recombinant bovine leukocyte antigens (BoLA) and recombinant bovine β2-microglobulin from which p-MHC class I tetramers can be derived in ~48 h. We validated a set of p-MHC class I tetramers against a panel of CTL lines specific to seven epitopes on five different antigens of Theileria parva, a protozoan pathogen causing the lethal bovine disease East Coast fever. One of the p-MHC class I tetramers was tested in ex vivo assays and we detected T. parva specific CTL in peripheral blood of cattle at day 15-17 post-immunization with a live parasite vaccine. The algorithm NetMHCpan predicted alternative epitope sequences for some of the T. parva CTL epitopes. Using an ELISA assay to measure peptide-BoLA monomer formation and p-MHC class I tetramers of new specificity, we demonstrate that a predicted alternative epitope Tp229-37 rather than the previously reported Tp227-37 epitope is the correct Tp2 epitope presented by BoLA-6*04101. We also verified the prediction by NetMHCpan that the Tp587-95 epitope reported as BoLA-T5 restricted can also be presented by BoLA-1*02301, a molecule similar in sequence to BoLA-T5. In addition, Tp587-95 specific bovine CTL were simultaneously stained by Tp5-BoLA-1*02301 and Tp5-BoLA-T5 tetramers suggesting that one T cell receptor can bind to two different BoLA MHC class I molecules presenting the Tp587-95 epitope and that these BoLA molecules fall into a single functional supertype.
dc.identifierhttps://hdl.handle.net/10568/41612
dc.identifier.urihttp://hdl.handle.net/123456789/148470
dc.languageen
dc.publisherSpringer
dc.rightsOpen Access
dc.sourceSvitek, N., Hansen, A.M., Steinaa, L., Saya, R., Awino, E., Nielsen, M., Buus, S. and Nene, V. 20014. Use of “one-pot, mix-and-read” peptide-MHC class I tetramers and predictive algorithms to improve detection of cytotoxic T lymphocyte responses in cattle. Veterinary Research 45:50.
dc.subjectcattle
dc.subjectvaccination
dc.subjectdiagnosis
dc.subjectlymphocytes
dc.titleUse of “one-pot, mix-and-read” peptide-MHC class I tetramers and predictive algorithms to improve detection of cytotoxic T lymphocyte responses in cattle
dc.typeJournal Article

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Instituto Internacional de Investigaciones Pecuarias (ILRI)