Figure 6
Inhibitory effects of 7-Keto cholesterol on antigen-specific CD4+T cell proliferation occur within first two hours of antigen receptor engagement. Lymph node cells were cultured with c-Ova323–339 peptide (A & B) or anti-CD3ε monoclonal antibody (C & D). 7-KC was added to the cultures either at 5 min (A & C) or at 24 hrs (B & D) after establishing cultures. Proliferating and non-proliferating cells were enumerated by two color staining with CFSE and anti-CD4-PE, and graphical representation of non-proliferating cells (CFSEhighest) CD4+ T cells in cultures is shown. These data were obtained from three independent experiments and for each time point was collected in triplicate. Statistical significance between treated and untreated groups was computed by one way ANOVA using the JMP program. Different lower case alphabet designations (a, b) above each treatment group (control, 35 μM 7KC, 17.5 μM 7KC, mβCD) indicates statistically significant difference (p <0.0001) with in each timed addition of 7KC and agonist used for T cell response (cOva323-339 5 minute/24 hr or anti-CD3ε 5 minute/24 hr). To examine statistical significance within a specific antigen receptor response (cOva323–339 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).