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. Author manuscript; available in PMC: 2013 Oct 31.
Published in final edited form as: J Immunol Methods. 2012 Jul 16;384(1-2):157–163. doi: 10.1016/j.jim.2012.07.002

Figure 3. Effect of ex vivo stimulation on detection of regulatory T cells.

Figure 3

Lymphocytes isolated from the gastric, splenic and mesenteric lymph node (MLN) compartments were cultured under either indirect (CD3/CD28, PMA/IONO) or direct stimulation (H. pylori lysate) and immunophenotype for CD25+ Foxp3+ regulatory T cells. Representative dot plots demonstrated that ex vivo stimulation yielded a decrease in the percentage of regulatory T cells regardless of the compartment the cells originated from. (A) This observation was quantified, and indicated that ex vivo stimulation significantly abrogates detection of CD4+ CD25+ Foxp3+ regulatory T-cells (B–D). Graphs represent the mean ± standard error of 4 mice pooled per experiment; results representative of 3 independent experiments. Groups were compared using one-way analysis of variance (ANOVA) followed post-hoc by Tukey multiple comparisons tests on log-transformed data. Significant differences are represented by the following *p<0.05, **p<0.01, ***p<0.001.