|ELNAGGAR, MAHMOUD - Washington State University|
|ABDELLRAZEQ, GABER - Washington State University|
|HULUBEI, VICTORIA - Washington State University|
|DAVIS, WILLIAM - Washington State University|
Submitted to: Developmental and Comparative Immunology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/3/2018
Publication Date: 4/5/2018
Citation: Elnaggar, M.M., Abdellrazeq, G.S., Dassanayake, R.P., Fry, L.M., Hulubei, V., Davis, W.C. 2018. Characterization of alpha beta and gamma delta T cell subsets expressing IL-17A in ruminants and swine. Developmental and Comparative Immunology. 85:115-124. https://doi.org/10.1016/j.dci.2018.04.003.
Interpretive Summary: Studies on the immune response in species other than humans and mice remain constrained by gaps in the monoclonal antibody reagents available for research, especially for cattle, sheep, goats, and swine. An objective of our ongoing research program has been to address this problem by developing monoclonal antibody where there is a critical need. In this context, we have developed a monoclonal antibody clone against a small protein called interleukin-17A (a type of a cytokine). This small protein promotes inflammation and is secreted by certain white blood cells in the blood upon stimulation with various stimuli including pathogens. This monoclonal antibody recognizes well conserved region in this small protein of cattle, sheep, goats, and pigs.
Technical Abstract: As part of our ongoing program to expand reagents available for research in cattle, we developed a monoclonal antibody (mAb) to bovine IL-17A, a multifunctional cytokine centrally involved in regulating innate and adaptive immune responses. Initial comparative studies demonstrated the mAb recognizes a conserved epitope expressed on orthologues of IL-17A in sheep, goats and pigs. Comparative flow cytometric analyses of lymphocyte subsets stimulated with Phorbol 12-myristate 13-acetate (PMA) and ionomycin revealed differences in the proportion of IL-17A+ CD4, CD8, and gamma delta T cells. Results in cattle showed the largest proportion of IL-17A+ cells were CD4+ followed by gamma delta+ and CD8+ T cells. Further analysis revealed the gamma delta T cell subset was comprised of WC1.1+, WC1.2+, and WC1- subsets. Analysis of the CD8+ subset revealed it was comprised of alpha beta and gamma delta T cell subsets. Results in sheep showed the largest proportion IL-17A+ cells were CD4+ followed by CD8+ alpha beta T cells and WC1+ gamma delta T cells. Results in goats showed the proportion of the IL-17A+ CD4, CD8, and gamma delta T cell subsets were similar. In contrast, a large proportion of the C8+ subset were gamma delta T cells. The majority of the IL-17A+ gamma delta T cells were WC1+. Results in swine showed the proportions of the IL-17A+ CD4, CD8, and gamma delta T cell subsets were similar. Comparison of expression of IL-17A with IFN-' revealed subsets co-expressed IL-17A and IFN-gamma in cattle, sheep, and goats. The new mAb expands opportunities for research in ruminants and swine.