Figure 7. T-cells from combination therapy treated mice produce IL-4 in response to proinsulin peptide stimulation.

At 60-days post anti-CD20 mono- (MT, open bars) or combination therapy (CT, filled black bars), CD4+ T-cells were purified from treated non-diabetic NOD mice. Simultaneously, T-depleted splenocytes (TDS) were obtained from 8–10 week old NOD mice and used as APCs. 250,000 purified CD4+ T-cells were incubated with proinsulin peptide or a mutated insulin B:9–23 (B16:A) peptide in the presence of 5×10⁴ APCs. Following 3-day incubation, IL-4 production was determined by ELISpot assay as previously described [34]. Background (media) subtracted spot numbers are shown on the Y-axis. Increased IL-4 production in response to proinsulin peptide was seen in combination therapy treated mice. Representative means ± SEM data from one of two independent experiments with similar results are shown. Statistical analysis was performed using unpaired students t-test using Graphpad Prism software.