title: Enzyme-linked immunosorbent spot assay analysis of peripheral blood lymphocyte reactivity to donor HLA-DR peptides: potential novel assay for prediction of outcomes for renal transplant recipients.
abstract: Chronic allograft dysfunction, which is the most common cause of late allograft failure, is in part caused by an ongoing immune response orchestrated by T lymphocytes primed by the indirect pathway of allorecognition. The low frequencies of such T cells have made it difficult to study indirect alloreactivity by using currently available assays. The development of a sensitive, clinically useful method of measuring indirect alloreactivity among human renal transplant recipients was thus attempted. Furthermore, in a pilot immunologic study, the contribution of the indirect pathway was studied in two groups of renal transplant recipients, i.e., patients with no prior acute rejection episodes and stable renal function ("stable" patients) and patients with at least one previous episode of biopsy-proven acute rejection, who were thus at risk for the development of chronic rejection ("high-risk" patients). The frequencies of type 1 T helper (interferon-gamma-producing) and type 2 T helper (interleukin-5- and -10-producing) peripheral blood lymphocytes reactive with a panel of synthetic peptides (corresponding to sequences from donor HLA-DR molecules) were determined for renal transplant recipients and normal control subjects by using an enzyme-linked immunosorbent spot assay (ELISPOT). Among recipients of DR-mismatched allografts, a cut-off value of 60 interferon-gamma spots/10(6) cells significantly (P = 0.02) separated stable patients (creatinine concentration, 1.1 +/- 0.3 mg/dl) from high-risk patients (creatinine concentration, 2.3 +/- 1.7 mg/dl). This is the first demonstration that the enzyme-linked immunosorbent spot assay can be used to monitor indirect alloreactivity to donor HLA-DR peptides among renal transplant recipients. These data provide the rationale for the prospective study of indirect alloreactivity among transplant recipients, to allow predictions of which patients would be at risk for the development of chronic rejection and thus allow appropriate planning of future interventions.
authors.affiliation: *Laboratory of Immunogenetics and Transplantation, Brigham and Women's Hospital, Nephrology Division, Children's Hospital, Harvard Medical School, Boston, Massachusetts, and Cellular and Molecular Immunology Laboratory, Schepens Eye Research Institute, and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.
authors.forename: Nader
authors.initials: N
authors.lastname: Najafian
authors.affiliation: *Laboratory of Immunogenetics and Transplantation, Brigham and Women's Hospital, Nephrology Division, Children's Hospital, Harvard Medical School, Boston, Massachusetts, and Cellular and Molecular Immunology Laboratory, Schepens Eye Research Institute, and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.
authors.forename: Alan D
authors.initials: AD
authors.lastname: Salama
authors.affiliation: *Laboratory of Immunogenetics and Transplantation, Brigham and Women's Hospital, Nephrology Division, Children's Hospital, Harvard Medical School, Boston, Massachusetts, and Cellular and Molecular Immunology Laboratory, Schepens Eye Research Institute, and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.
authors.forename: Eugenia V
authors.initials: EV
authors.lastname: Fedoseyeva
authors.affiliation: *Laboratory of Immunogenetics and Transplantation, Brigham and Women's Hospital, Nephrology Division, Children's Hospital, Harvard Medical School, Boston, Massachusetts, and Cellular and Molecular Immunology Laboratory, Schepens Eye Research Institute, and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.
authors.forename: Gilles
authors.initials: G
authors.lastname: Benichou
authors.affiliation: *Laboratory of Immunogenetics and Transplantation, Brigham and Women's Hospital, Nephrology Division, Children's Hospital, Harvard Medical School, Boston, Massachusetts, and Cellular and Molecular Immunology Laboratory, Schepens Eye Research Institute, and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.
authors.forename: Mohamed H
authors.initials: MH
authors.lastname: Sayegh
mesh_terms: D001773:Blood Cells / Q000276:immunology*; D002908:Chronic Disease; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006084:Graft Rejection / Q000209:etiology; D006684:HLA-DR Antigens / Q000276:immunology*; D006650:Histocompatibility Testing; D006801:Humans; D016030:Kidney Transplantation / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D010865:Pilot Projects; D011379:Prognosis; D012016:Reference Values; D015203:Reproducibility of Results; D012307:Risk Factors; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology; D014019:Tissue Donors; D016896:Treatment Outcome
chemical_list: D006684:HLA-DR Antigens; D010446:Peptide Fragments