Statistics

Multiple Student’s t tests with FDR correction were used to determine the statistical significance for all the results presented, except Supplementary Figure 1 where a Log-rank (Mantel-Cox) test and chi-square test was used and Figure 4A where two-way ANOVA was used. Statistical analysis was conducted using Prism 8.

2H6 TCR transgene strongly reduces antigen presentation by insulin-reactive VH125 B cells and increases anti-inflammatory TGFβ secretion. Splenic B cells were negatively isolated and cultured with bead-purified CD4+ T cells from 8-week-old mice in a 1:1 ratio. Cells were co-cultured with denatured insulin or insulin B chain 12-25 peptide at various concentrations for 48 h prior to supernatant collection and ³H-thymidine addition. Proliferation was measured by ³H-thymidine incorporation and plotted as ΔCPM, where the background CPM (cells without antigen) was subtracted from the CPM of cells with antigen. (A, B) 2H6 CD4+ T cells isolated from 2H6 NOD mice (A) or 2H6 T cells isolated from 2H6VH125 NOD mice (B) co-cultured with B cells from various donors and denatured insulin. (C, D) 2H6 CD4+ T cells isolated from 2H6 NOD mice (C) or 2H6 T cells isolated from 2H6VH125 NOD mice (D) co-cultured with B cells from various donors and insulin B chain 12–25 peptide. (E–H) TGFβ measurements from the supernatants of the respective cultures in (A–D). Data were generated from three independent experiments (n = 2–3 mice/experiment). Data from NOD mice are shown as a non-transgenic mouse comparison. Data were assessed for significance using two-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.0001.