Supplementary Figure 15: Diminished tolerance to self-antigen caused by the I325N A20 allele.

InsHEL-transgenic mice or non-transgenic controls, both with wild-type A20 in pancreatic islets, were transplanted with a congenic bone marrow mixture from Tnfaip3^+/+ CD45.1⁺ TCR^3A9-transgenic donors, to serve as an internal control, and CD45.2 TCR^3A9-transgenic donors with either Tnfaip3^I325N/I325N (black circles) or Tnfaip3^+/+ (grey circles) genotype. (a) Frequency and mean of TCR^3A9 positive CD4⁺ T cells among CD45.2⁺ Tnfaip3^+/+ (grey circles) or Tnfaip3^I325N/I325N (black circles) lymphocytes in pancreatic lymph nodes. Test recipients expressed the insHEL antigen in pancreatic islets (insHEL-transgenic), while parallel non-transgenic recipients lacked the autoantigen. (b) Percentage and mean of CD45.2⁺ Tnfaip3^+/+ (grey circles) or Tnfaip3^I325N/I325N (black circles) cells among TCR^3A9+ CD4⁺ T cells in spleen. (c, d) Analysis of insHEL transgenic recipients. Relative mean fluorescence intensity (MFI) of staining for cell surface MHC class II or CD86 on CD45.2⁺ Tnfaip3^+/+ (grey circles) or Tnfaip3^I325N/I325N (black circles) B cells (c) or CD11c⁺ dendritic cells (d), compared to CD45.1⁺ Tnfaip3^+/+ B cells or dendritic cells in the same sample. (e) Frequency of germinal center (GC) B cells among CD45.2+ B cells (black circles) compared to the frequency among CD45.1⁺ Tnfaip3^+/+ B cells in the same animal (linked by lines). Left panel, CD45.2 Tnfaip3^+/+ marrow donor; right panel, CD45.2 Tnfaip3^I325N/I325N marrow donor. In (c-e), each symbol, or pair of symbols joined by a line, represents one chimeric mouse, all analyzed in a single experiment. Statistical analysis by 2-tailed Student’s t-tests: *P < 0.05; **P < 0.01; ***P < 0.001.