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

Nicholas Svitek, Andreas Martin Hansen, Lucilla Steinaa, Rosemary Saya, Elias Awino, Morten Nielsen, Søren Buus, Vishvanath Nene

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    Abstract

    Peptide-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 beta 2-microglobulin from which p-MHC class I tetramers can be derived in similar to 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 Tp2(29-37) rather than the previously reported Tp2(27-37) epitope is the correct Tp2 epitope presented by BoLA-6*04101. We also verified the prediction by NetMHCpan that the Tp5(87-95) epitope reported as BoLA-T5 restricted can also be presented by BoLA-1*02301, a molecule similar in sequence to BoLA-T5. In addition, Tp5(87-95) 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 Tp5(87-95) epitope and that these BoLA molecules fall into a single functional supertype.
    Original languageEnglish
    JournalVeterinary Research
    Volume45
    Issue number1
    Number of pages16
    ISSN0928-4249
    DOIs
    Publication statusPublished - 2014

    Bibliographical note

    © 2014 Svitek et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.

    Keywords

    • VETERINARY
    • EAST-COAST FEVER
    • THEILERIA-PARVA
    • ANTIGEN PRESENTATION
    • BINDING PREDICTION
    • IMMUNE-RESPONSES
    • CELL RESPONSES
    • MOLECULES
    • SPECIFICITY
    • SEQUENCE
    • PROTEIN

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