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Table 2 T hybridoma response to single site (alanine or aspartic acid) substituted altered peptide ligands of HC gp-39(263–275)

From: Identification of an altered peptide ligand based on the endogenously presented, rheumatoid arthritis-associated, human cartilage glycoprotein-39(263–275) epitope: an MHC anchor variant peptide for immune modulation

     Hybridoma response (SI)
Compound HC gp-39 Sequence HLA-DR binding IC50 (μM) 5G11 8B12 14G11 20H5
1 263–275 RSFTLASSETGVG 0.08 17 39 30 5
10 R263A1 ASFTLASSETGVG 0.10 23 42 28 5
11 S264A2 RAFTLASSETGVG 0.10 21 43 32 5
12 F265A3 RSATLASSETGVG >35 2 6 2 4
13 T266A4 RSFALASSETGVG 0.06 7 14 1 5
14 L267A5 RSFTAASSETGVG 0.08 4 1 1 4
15 A268D6 RSFTLDSSETGVG 0.24 1 1 1 2
16 S269A7 RSFTLAASETGVG 0.05 2 1 1 2
17 S270A8 RSFTLASAETGVG 0.05 21 29 27 5
18 E271A9 RSFTLASSATGVG 0.04 1 1 1 2
19 T272A10 RSFTLASSEAGVG 0.04 1 4 1 1
20 G273A11 RSFTLASSETAVG 0.05 7 32 6 5
21 V274A12 RSFTLASSETGAG 0.06 26 40 30 5
22 G275A13 RSFTLASSETGVA 0.05 17 30 20 5
23 A3/A8 RSATLASAETGVG >36 ND ND ND ND
  1. Binding of peptides to HLA-DR4 (B1*0401) was determined in a competition binding assay. IC50 = 50% inhibitory concentration. IC50 < 1 μM = high affinity binder; IC50 between 1 and 10 μM = good binder; IC50 between 10 and 100 μM = intermediate binder. The HLA-DRB1*0401 expressing B lymphoblastoid cell line was used as a source of antigen-presenting cells. Background values for hybridomas 5G11, 8B12, 14G11 and 20H5 were 17,238, 8,773, 10,131 and 290,666 mean fluorescence units/counts, respectively. The standard deviation of measurements did not exceed 15%. Hybridoma IL-2 response is expressed as stimulation index (SI) calculated as the ratio of mean fluorescence counts of antigen stimulated cultures and control cultures. Values in bold are considered positive (SI > 2). Data are representative of at least three independent experiments. ND, not determined.