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Title: Differences in leukocyte differentiation molecule abundances on domestic sheep (Ovis aries) and bighorn sheep (Ovis canadensis) neutrophils identified by flow cytometry

Author
item Highland, Margaret
item Schneider, David
item White, Stephen
item MADSEN-BOUTERSE, SALLY - Washington State University
item Knowles Jr, Donald
item DAVIS, WILLIAM - Washington State University

Submitted to: Comparative Immunology Microbiology and Infectious Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/13/2016
Publication Date: 4/14/2016
Citation: Highland, M.A., Schneider, D.A., White, S.N., Madsen-Bouterse, S.A., Knowles Jr, D.P., Davis, W.C. 2016. Differences in leukocyte differentiation molecule abundances on domestic sheep (Ovis aries) and bighorn sheep (Ovis canadensis) neutrophils identified by flow cytometry . Comparative Immunology Microbiology and Infectious Diseases. doi: 10.1016/j.cimid.2016.04.006.

Interpretive Summary: Bighorn and domestic sheep pneumonia is production and population limiting. Previous work has indicated bighorn sheep are more susceptible to certain pathogens and therefore pneumonia compared to domestic sheep. An important part of the innate immune system is the neutrophil and the neutrophil is important in pneumonia of bighorn and domestic sheep. This work tested for a difference in certain neutrophil markers that predict responses to pathogens. A difference in markers was found between the two species. Additional work will determine the importance of these markers in response to specific pathogens.

Technical Abstract: Although both domestic sheep (DS) and bighorn sheep (BHS) are affected by similar respiratory bacterial pathogens, experimental and field data indicate BHS are more susceptible to pneumonia. Cross-reactive monoclonal antibodies (mAbs) for use in flow cytometry (FC) are valuable reagents for interspecies comparative immune system analyses. This study describes cross-reactive mAbs that recognize leukocyte differentiation molecules (LDMs) and major histocompatibility complex antigens on DS and BHS leukocytes. Characterization of multichannel eosinophil autofluorescence in this study permitted cell-type specific gating of granulocytes for evaluating LDMs, specifically on neutrophils, by single-label FC. Evaluation of relative abundances of LDMs by flow cytometry revealed greater CD11a, CD11b, CD18 (ß2 integrins) and CD 172a (SIRPa) on DS neutrophils and greater CD14 (lipopolysaccharide receptor) on BHS neutrophils. Greater CD25 (IL-2) was identified on BHS lymphocytes following Concavalin A stimulation. While DS and BHS have similar total peripheral blood leukocyte counts, BHS have proportionately more neutrophils.