Figure 4
Inhibition of antigen-specific proliferation of CD4+T cells by 7-Keto cholesterol in dose-dependent fashion. A. Lymph node cells were cultured with c-Ova323–339 stimulatory or c-Ova324–334 control peptide (A & B) or anti-CD3ε monoclonal antibody (C) either in the absence or presence of 7-KC. Non-dividing cells were enumerated by two color staining with CFSE (X-axis) and anti-CD4-PE (Y-axis), a representative experiment is shown (A). Responsiveness to stimulation through the antigen receptor was quantified by enumerating non-proliferating cells (upper right quadrant) in these cultures (B & C). These data were obtained from three independent experiments. Statistical significance between treated and untreated groups was computed by one way ANOVA using JMP program. 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 <0.001) in the presence of either cOva323–339 peptide or anti-CD3ε antibody. To examine statistical significance within a specific antigen receptor response (cOva323–339 and 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).