Peptidomic analysis of type-1 diabetes associated HLA-DQ molecules and impact of HLA-DM editing (APP5P.108)

Abstract

Abstract HLA-DM is critical in editing peptide repertoires presented by APC, thus influencing the selection/activation of CD4+ T cells. We previously reported that type-1 diabetes (T1D) associated HLA-DQ2, 8 & DQ2/8 molecules (DQs) are relatively resistant to DM editing of the CLIP peptides. In this study, we further studied DM sensitivity of T1D-associated DQs by peptidomic analysis. Compared to a T1D-protective DQ6, the percentages of shared peptides among T1D-associated DQs were significantly higher. There were greater numbers of nested peptide sets and long peptides (>20 aa) eluted from T1D-associated DQs. Predicted peptide binding motifs of DQ2, 8 & 2/8 shared charged anchor residues, while hydrophobic anchors were present in DQ6 peptides. Competition binding assays comparing the eluted peptides from DM sensitive, dependent, or resistant groups showed that the binding affinities of the DM sensitive peptides were significantly lower than that of CLIP1 peptide. The increased sDQ8-CLIP1 complex stability, with the intrinsic half-life of 58 h measured by fluorescence polarization assay, was 8 times longer than that of sDQ6-CLIP1 (7 h), further supporting the DM resistance of T1D associated DQs. Collectively, T1D associated DQs share distinct characteristics, including binding of peptides from known T1D auto-antigens. Our new data further support the hypothesis that DM resistance in T1D associated DQs affects the peptide repertoires, thus contributing to the pathogenesis of T1D.