Figure 7

Effect of 7-KC on the proliferation of CD4⁺T cell and APC co-cultures where only CD4⁺T cells were pre-treated with 7-KC. Purified CD4⁺ T cells were incubated with 7-KC for 30 minutes at 37°C and excess of 7-KC was removed by washing with wash media. CD4⁺ T cells were stained with CFSE prior to being combined with APCs. 7-KC treated and mock-treated (labeled as control) CD4⁺ T cells were cultured with c-Ova_323–339 peptide for 72 hrs and stained with anti-CD4⁺ PE enumeration of CD4⁺ T cells only as described in materials and methods section. Proliferating and non-proliferating cells were enumerated by two color staining with CFSE (X-axis) and anti-CD4-PE (Y-axis) and graphical representation of non-proliferating cells (CFSE^highest) CD4⁺ T cells in cultures is shown. These data were obtained from three independent experiments and each 7-KC concentration was carried out in triplicates. Statistical significance between treated and untreated groups was computed by one way ANOVA using JMP program. Different small alphabet designations above each bar indicate statistically significant differences (p < 0.0221). Different lower case alphabet designations (a, b, c) above each treatment group (control, 35 μM 7KC, 17.5 μM 7KC, mβCD) indicates statistically significant difference (p < 0.0221) in response to either cOva_323–339 peptide or anti-CD3ε antibody. To examine statistical significance within a specific antigen receptor response (cOva_323–339 or anti-CD3ε), each treatment group (e.g., 7KC) is compared with other (all possible combinations). Similar lower case alphabet designation indicates lack of statistical significance (p > 0.05). Double designation (ab) indicates lack of statistical significance with experimental treatments with designations “a” as well as “b”.