Projects per year
Abstract / Description of output
The development of methods to detect cytokine expression by T cell subsets in ruminants is fundamental to strategic development of new livestock vaccines for prevention of infectious diseases. It has been possible to detect T cell expression of IFN-γ, IL-4 and IL-10 in ruminants for many years but methods to detect expression of IL-17A are relatively limited. To address this gap in capability we have cloned bovine and ovine IL-17A cDNAs and expressed biologically-active recombinant proteins in Chinese Hamster Ovary (CHO) cells. We used the transfected CHO cells to screen commercially-available antibodies for their ability to detect IL-17A expression intracellularly and in culture supernates. We demonstrate that an ELISA for bovine IL-17A detects native ovine IL-17A. Moreover, the constituent polyclonal antibodies (pabs) in the ELISA were used to enumerate peripheral blood mononuclear cells (PBMC) expressing IL-17A from cattle and sheep by ELISpot. We identified two monoclonal antibodies (mabs) that detect recombinant intracellular IL-17A in CHO cells by flow cytometry. One of these mabs was used to detect native intracellular IL-17A expression in PBMC in conjunction with cell surface phenotyping mabs (CD4+ve, CD8+ve and Workshop Cluster 1 [WC-1]+ve gamma-delta [γδ]) we show that distinct T cell subsets in cattle and sheep can express IL-17A following activation. These novel techniques provide a solid basis to investigate IL-17A expression and define specific CD4+ve T cell subset activation in ruminants.
Original language | English |
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Article number | 20 |
Journal | Veterinary Research |
Volume | 48 |
Issue number | 1 |
Early online date | 8 Apr 2017 |
DOIs | |
Publication status | E-pub ahead of print - 8 Apr 2017 |
Keywords / Materials (for Non-textual outputs)
- ovine
- bovine
- IL-17A
- T cell subsets
- intracellular cytokine staining
- ELISA
- ELISpot
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Dive into the research topics of 'Enhancing the toolbox to study IL-17A in cattle and sheep'. Together they form a unique fingerprint.Projects
- 2 Finished
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Innate immunity and endemic diseases in livestock species
Collie, D., Beard, P., Bishop, S., Bronsvoort, M., Burt, D., Fitzgerald, R., Freeman, T., Gally, D., Gill, A., Glass, E., Hocking, P., Hope, J., Hume, D., Kaiser, P., Mabbott, N., McLachlan, G., Morrison, L., Stevens, J., Stevens, M. & Watson, M.
1/04/12 → 31/03/17
Project: Research
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The route to identification of the
Glass, E., Hope, J. & Entrican, G.
1/02/12 → 31/01/15
Project: Research