Figure 3: NOD mice present B and T autoimmune responses against mIL-2.
(a,b) The number of anti-mIL-2 secreting cells in fresh splenocytes and bone marrow cells (plasma cells, a) or in 5-days CpG pre-activated splenocytes (memory B cells, b) from 10–18 week-old female B6 or pre-diabetic NOD mice was quantified by ELISPOT. (c) IFN-γ production by 10–18-week-old female B6 (n=3) or pre-diabetic NOD (n=7) splenocytes was quantified in culture supernatants by CBA after 72 h of stimulation with DMSO, mIL-2 peptides that gave a positive response in the initial screen (3 and 10 μM of each peptide), P31 peptide (3 and 10 μM) or aCD3-CD28 coated beads (ratio 1bead:1cell). (d) mIL-2 amino acid sequence. The regions corresponding to peptides that gave positive results in the initial screen are underlined and the immunodominant peptides (mIL-210-24 and mIL-267-81) are in bold. The mIL-2 signal peptide is indicated by a rectangle. (a–c) Symbols represent individual mice and horizontal bars are the medians. Data are cumulative of at least two independent experiments. (a,b) *P<0.05 (Kruskal-Wallis test with Dunn’s multiple comparisons test).
