2016 ACVIM Forum Research Abstract Program

PBMCs from the same 10 horses were cultured with each of the following, no stimulation, conA, and levamisole with and without conA. To determine proliferation of each specific subset, cells were labelled with a fluorescent dye (CellTrace Violet, ThermoFisher Scientific, C34557). Proliferation of each subset was determined based on dye dilution using flow cytometry (FacsAria Flow Cytometer). To determine the ability of levamisole to upregulate or alter the immune response, immune subsets were identified (CD4, CD8, CD21, CD172a, CD14) using fluorescent labelled antibodies. Activation was assessed for macrophages and DCs using MHC class II and CD86 expression. Induction of T-regs was based on CD4, foxp3 expression. Specific immune phenotypes were determined based on intracellular cytokine expression of specific subsets (M1 DC1 CD4 Th1 CD8: IFN-gamma) versus (M2 DC2 CD4 Th2: IL4) versus (CD4 T-reg: IL-10). Significant differences in response were determined using a mixed model ANOVA with significance set at P < 0.05.