Figure 6
Inhibition of IFN-γ, IL-2, IL-6 and TNF-α cytokine release from MLRs by Didox and Trimidox. Major antigen mismatch (filled square): Briefly, 2 × 105 T-cells were purified from spleens of C57BL6 mice (responder cells) which were mixed with 4 × 105 non T-cells (stimulator cells) obtained from BALB/c mice and exposed to 3000 rads of γ-radiation. Minor antigen mismatch (open square): Briefly, 2 × 105 T-cells were purified from the spleens of B10.D2 mice (responder cells) which were mixed with 4 × 105 non T-cells (stimulator cells) obtained from BALB/c mice and previously exposed to 3000 rads of γ-radiation. Either PBS or several concentrations of Didox or Trimidox (25 μM- 100 μM) was added to RPMI 1640 growth media supplemented with 10% fetal bovine serum, 2-mercaptoethanol (50 μM)and 1% penicillin streptomycin. The plates were then incubated at 37C and 5% CO2 for 6 days, after which the levels of IFN-γ, IL-2 and IL-6 were determined using the Searchlight multiplex assay system. NC: Normal control, AC: Activated control. The results shown represent data obtained in triplicate from two independent experiments (n = 3). * p < 0.05, ** p < 0.001 indicates a significant difference compared to activated control (AC), ANOVA + the Bonferroni test.