HLA-B7 beta-pleated sheet-derived synthetic peptides are immunodominant T-cell epitopes regulating alloresponses.
Chronic rejection of transplanted allografts is the major cause of graft loss after clinical solid organ transplantation. Recent data link the indirect presentation of allopeptides to chronic graft loss; thus, identification of immunodominant epitopes on major histocompatibility complex (MHC) antigens could significantly contribute to establishing novel ways for monitoring and managing chronic rejection. Here, we show that synthetic allo-MHC-derived peptides covering the polymorphic region 56 to 120 of HLA-B7 modulate alloresponses. In particular, the 2 beta-pleated sheet-derived peptides covering residues 91 to 105 and 96 to 120, respectively, but not sequences from the alpha1 helix, were presented by autologous peripheral blood lymphocytes to induce T-cell proliferation. In addition, the 2 beta-pleated sheet-derived peptides and the alpha1-derived peptide residues 60 to 75 abrogated lysis of HLA-B7 target cells by anti-HLA-B7 cytotoxic T lymphocytes (CTLs). Although most residues between 91 and 120 are normally not directly accessible to T cells, our results indicate that peptides derived from the lower surface of the peptide-binding groove of HLA-B7 are immunodominant in HLA-B7 alloresponses. To characterize the binding and stability of allopeptides to T cells, the 62-70 peptide-derived from the 60-75 allopeptide that blocked cytotoxicity of anti-HLA-B7 CTL-was synthesized and coupled with fluorescein isothiocyanate. The peptide specifically labeled anti-B7 CTL, but not anti-HLA-A2 CTL as measured by flow cytometry. Peptide binding to CTL was specific at 4 degrees C and remained stable for 12 hours, whereas it remained stable for less than 2 hours at 37 degrees C. These studies allow the identification of HLA-B7 T-cell epitopes and reveal for the first time a novel, previously unrecognized application of synthetic HLA-derived allopeptides to visualize alloreactive T cells.
Freese
Anja
A
Department of Internal Medicine, University of Iowa Hospitals and Clinics and VA Medical Center, Iowa City 52242, USA.
Zavazava
Nicholas
N
D000595:Amino Acid Sequence; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D015793:HLA-B7 Antigen / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes; D008285:Major Histocompatibility Complex / Q000276:immunology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D013696:Temperature
D018984:Epitopes, T-Lymphocyte; D015793:HLA-B7 Antigen; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D016650:Fluorescein-5-isothiocyanate