title: Chemokine receptor 7 (CCR7)-expression and IFNγ production define vaccine-specific canine T-cell subsets.
abstract: Canines suffer from and serve as strong translational animals models for many immunological disorders and infectious diseases. Routine vaccination has been a mainstay of protecting dogs through the stimulation of robust antibody responses and expansion of memory T-cell populations. Commercially available reagents and described techniques are limited for identifying and characterizing canine T-cell subsets and evaluating T-cell-specific effector function. To define reagents for delineating naïve versus activated T-cells and identify antigen-specific T-cells, we tested anti-human and anti-bovine T-cell specific cell surface marker reagents for cross-reactivity with canine peripheral blood mononuclear cells (PBMCs. Both CD4(+) and CD8(+) T-cells from healthy canine donors showed reactivity to CCL19-Ig, a CCR7 ligand, and coexpression with CD62L. An in vitro stimulation with concanavalin A validated downregulation of CCR7 and CD62L expression on stimulated healthy control PBMCs, consistent with an activated T-cell phenotype. Anti-IFNγ antibodies identified antigen-specific IFNγ-producing CD4(+) and CD8(+) T-cells upon in vitro vaccine antigen PBMC stimulation. PBMC isolation within 24h of sample collection allowed for efficienT-cell recovery and accurate T-cell effector function characterization. These data provide a reagent and techniques platform via flow cytometry for identifying canine T-cell subsets and characterizing circulating antigen-specific canine T-cells for potential use in diagnostic and field settings.
authors.affiliation: Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA; Department of Infectious Diseases, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Ashley N
authors.initials: AN
authors.lastname: Hartley
authors.affiliation: Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, 30602, USA; Department of Cellular Biology, University of Georgia, Athens, GA, 30602, USA. Electronic address: tarleton@uga.edu.
authors.forename: Rick L
authors.initials: RL
authors.lastname: Tarleton
mesh_terms: D000818:Animals; D004285:Dogs; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis*; D019041:L-Selectin / Q000032:analysis; D054400:Receptors, CCR7 / Q000032:analysis*; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D014612:Vaccines / Q000276:immunology*
chemical_list: D054400:Receptors, CCR7; D014612:Vaccines; D019041:L-Selectin; D007371:Interferon-gamma
keywords: CCR7; CD62L; Dog; Flow cytometry; T-cell; Vaccine