Naturally processed peptides spanning the HPA-1a polymorphism are efficiently generated and displayed from platelet glycoprotein by HLA-DRB3*0101–positive antigen-presenting cells

Abstract

AbstractIn neonatal alloimmune thrombocytopenia, almost all human platelet antigen (HPA)–1b1b mothers who produce anti–HPA-1a antibody through carrying an HPA-1a fetus are human histocompatibility leukocyte antigen (HLA)–DRB3*0101 positive. It is predicted that the HPA-1a Leu33 polymorphism forms part of an HLA-DRB3*0101–restricted T-helper epitope, and acts as an anchor residue for binding this class II molecule. However, it is not known whether any corresponding peptides are naturally processed and presented from platelet glycoprotein. In this study, peptides displayed by a homozygous HLA-DRB3*0101 antigen-presenting cell line were identified after pulsing with recombinant HPA-1a (Leu33 plexin-semaphorin-integrin domain). The peptides were eluted from HLA-DR molecules, fractionated by high performance liquid chromatography, and analyzed by tandem mass spectrometry. A “nested set” of naturally presented HPA-1a–derived peptides, each containing the Trp25-Leu33 core epitope, was identified, with the most abundant member being the 16-mer Met22-Arg37. These peptides may provide the basis for novel treatments to tolerize the corresponding T-helper response in women at risk of neonatal alloimmune thrombocytopenia.