Extended Data Fig. 4. (see also Fig. 3). HLA-DR restriction and response to autoantigens of TCC88.
a, Several of the isolated TCCs were stimulated with irradiated BLS cells expressing single HLA-DR molecules of patient 1635WI (BLS-DRB1*03:01, BLS-DRB1*04:02, BLS-DRB3*02:02 and BLS-DRB4*01:01) as APCs and dose titration of SIN3A* peptide (10, 1, 0.1, 0.01 and 0.001 µM). Proliferation was measured using 3H-thymidine incorporation after 3 days. Data represents mean SI of 5 replicate wells ± SEM). Several TCCs did not respond to peptides presented on any of HLA-DR molecules (data not shown). b-c, Median IFN-γ response of three independent experiments (transformed to log base 10 values) of TCC88 stimulated via ps-SCL was measured and depicted for 20 L-amino acids fixed in all positions of a 10-mer peptide. c, IFN-γ values were depicted in a table format (scoring matrix) and represent the relative stimulatory potency of each L-amino acid in different positions of the decamers. d, Relationship between predicted score and stimulatory index (SI) was established by testing potential cognate antigens within the predicted score range of 60–100% of the maximal score with TCC88. e–f, TCC88 was tested with pools of overlapping peptides (including 260 peptides) covering brain, pancreas and skin autoantigens (Supplementary Table 11) including peptide pools covering myelin basic protein (MBP pools), myelin oligodendrocyte glycoprotein (MOG pools), myelin proteolipid protein (PLP pool contains 5 immunodominant PLP peptides), RAS guanyl-releasing protein 2 (RASGRP2 pools),GDP-L-fucose synthase (TSTA3 pools), MELAN-A, tyrosinase (TYR), premelanosome protein (PMEL), insulin, SLC30A8 (ZnT8) and islet amyloid polypeptide (IAPP). TCC88 was stimulated with peptide-pulsed irradiated BLS-DRB3*02:02 cells for 3 days. Proliferation was measured using 3H-thymidine incorporation (data represents mean value of 3 replicate wells ±SEM). f, Response of TCC88 to unmutated SIN3A peptide was compared to SIN3A* at different concentrations including 10, 1, 0.1, 0.01 and 0.001 µM.