The Peptide Repertoire of HLA‐B27 may include Ligands with Lysine at P2 Anchor Position

Abstract The HLA‐B*27 peptidome has drawn significant attention due to the genetic association between some of the HLA‐B*27 alleles and the inflammatory rheumatic disease ankylosing spondylitis (AS), for which a comprehensive biological explanation is still lacking. This study aims to expand the known limits of the HLA‐B*27 peptidome to facilitate selection and testing of new peptides, possibly involved in the disease. The HLA peptidomes of HeLa and C1R cell lines stably transfected with the AS‐associated HLA‐B*27:05 allele, the nonassociated HLA‐B*27:09 allele, or their cysteine 67 to serine mutants (C67S), are analyzed on a very large scale. In addition, the peptidomes of HLA‐B*27:05 and HLA‐B*27:05‐C67S are analyzed from the spleens of rats transgenic for these alleles. The results indicate that C67S mutation increases the percentage of peptides with glutamine or lysine at their P2 position (P2‐Lys), in both HLA‐B*27:05 and HLA‐B*27:09. Furthermore, a small fraction of HLA‐B*27 peptides contains lysine at their second position (P2), in addition to the more commonly found peptides with arginine (P2‐Arg) or the less common glutamine (P2‐Gln) located at this anchor position. Overall these data indicate that peptides with P2‐Lys should be considered as real ligands of HLA‐B*27 molecules and taken into account while looking for putative peptides implicated in the AS.