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An analysis gate was set on the lymphocyte population based on the forward and side scatter plot. Firstly, CD3+-stained cells falling within the gated area were identified. The gated population was further analyzed for the identification of CD3+CD4+ (CD4+) or CD3+CD8+ (CD8+). Previous studies have shown that the stimulation with PMA and calcium ionophore downregulates surface CD4 expression on T cells, which may make it difficult to identify CD4+ T cells . Thus, the CD4+ population was also analyzed by gating on the CD3+CD8- population. Cells positive for IL-10 were expressed as a percentage of each subset. The cut-off level for definition of positive cells was set so that less than 2% of isotype antibody-stained cells were positive. Then, the frequency of true-positive cells was obtained by subtracting the value of isotype control from the value of sample stained with anti-IL-10 mAb. Statistical analyis Statistical analysis was completed using StatView J4.02 (Abacus Concepts Inc, USA). Data were expressed as medians and interquartile ranges. When multiple comparisons were made among groups, significant differences were assessed using the Kruskal-Wallis test, followed by the Mann-Whitney U test. Wilcoxon signed rank test was used for within-group comparisons. Values of P less than 0.05 were accepted as statistically significant.