Fig. 2

LTC treatment of dog macrophages stimulates TNFα production and upregulation of MHCII expression. Whole blood was obtained from 3 dogs and processed to generate PBMCs, as noted in Fig. 1. Monocytes were enriched by plastic adherence of PBMC to triplicate wells of a 24-well plates and differentiated to macrophages by incubation in M-CSF, as noted in Methods. Macrophages were ether untreated or treated for 18 h with either 1 ul/ml, 5 ul/ml, 10 ul/ml of LTC or 10 ng/ml canine IFNg or 500 ng/ml LPS. Secretion of TNFα in supernatants was assessed by canine TNFα ELISA (panel a). Expression of MHCII was assessed by flow cytometry (panel b), as noted in Methods. Data were analyzed using one-way ANOVA with multiple means comparisons. (*, P ≤ 0.05, **, P ≤ 0.01, P ≤ 0.001). These results are representative of a total of 3 separate and independent experiments