Structural aspects of human leukocyte antigen class I epitopes detected by human monoclonal antibodies.
This study addresses the concept that human leukocyte antigen (HLA) class I-specific alloantibodies are specific for epitopes that correspond to HLAMatchmaker-defined eplets. Eplets are essential parts of so-called structural epitopes that make contact with the 6 complementarity determining regions of an antibody. From published molecular models of crystallized protein antigen-antibody complexes, we have calculated that contact residues on structural HLA epitopes should reside within a 15-Å radius of a mismatched eplet. This study addresses the structural basis of high-frequency HLA class I epitopes reacting with human monoclonal antibodies (mAbs) derived from women sensitized during pregnancy. All mAbs were tested in Luminex assays with single HLA allele panels. The HLAMatchmaker algorithm was used to determine their specificity in context with eplet sharing between the immunizing allele and antibody-reactive alleles. To assess the autoreactive B cell origin of these antibodies, we have applied the recently developed nonself-self paradigm of epitope immunogenicity to analyze residue differences between the immunizer and the alleles of the antibody producer. A total of 9 mAbs were specific for epitopes associated with the 41T, 80NRG, 163LW, 69AA, or 80ERILR eplets. In each case, the immunizing allele had within 15 Å of the mismatched eplet, no residue differences with 1 of the alleles of the antibody producer. This observation is consistent with the concept that these mAbs originated from B cells with self HLA immunoglobulin receptors. Eplet-carrying alleles exhibited different levels of reactivity, which, when compared with the immunizing allele, ranged from high to intermediate to very low. In many cases, lower reactivities were associated with differences from self to nonself residues in surface locations within 15 Å of the specific eplet. Apparently, such locations may serve as critical contact sites for the antibody. In other cases, other residue differences did not appear to affect binding with the antibody, suggesting that these locations do not play a major role in antibody binding. For these mAbs we did not obtain convincing evidence that residue differences in hidden positions below the molecular surface had significant effects on antibody binding. These findings have increased our understanding of the structural basis of the immunogenicity and antigenicity of HLA class I epitopes and provide a basis for interpreting HLA antibody reactivity patterns in Luminex assays with single alleles.
Division of Transplantation Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA 15261, USA. Duquesnoyr@upmc.edu
Rene J
RJ
Duquesnoy
Marilyn
M
Marrari
Arend
A
Mulder
Frans H J
FH
Claas
Justin
J
Mostecki
Ivan
I
Balazs
D000465:Algorithms; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry* / Q000276:immunology; D019295:Computational Biology; D003460:Crystallization; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry* / Q000276:immunology; D005260:Female; D006680:HLA Antigens / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D006825:Hybridomas; D007114:Immunization; D008956:Models, Chemical; D011247:Pregnancy; D011485:Protein Binding; D011487:Protein Conformation; D012984:Software
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D018985:Epitopes, B-Lymphocyte; D006680:HLA Antigens