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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
veterinary
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, Volume 45, No. 1, Article 50, Year 2014
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Description
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 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 Tp2 27-37 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. © 2014 Svitek et al.; licensee BioMed Central Ltd.
Authors & Co-Authors
Svitek, Nicholas
Kenya, Nairobi
International Livestock Research Institute Nairobi
Hansen, Andreas Martin
Denmark, Copenhagen
Københavns Universitet
Steinaa, Lucilla
Kenya, Nairobi
International Livestock Research Institute Nairobi
Saya, Rosemary M.
Kenya, Nairobi
International Livestock Research Institute Nairobi
Awino, Elias
Kenya, Nairobi
International Livestock Research Institute Nairobi
Nielsen, Morten
Denmark, Lyngby
Technical University of Denmark
Argentina, San Martin
Universidad Nacional de San Martín
Buus, Soren
Denmark, Copenhagen
Københavns Universitet
Nene, Vishvanath M.
Kenya, Nairobi
International Livestock Research Institute Nairobi
Statistics
Citations: 30
Authors: 8
Affiliations: 4
Identifiers
Doi:
10.1186/1297-9716-45-50
ISSN:
09284249
e-ISSN:
12979716
Research Areas
Maternal And Child Health