title: Detection of anti-HLA antibodies with flow cytometry in needle core biopsies of renal transplants recipients with chronic allograft nephropathy.
abstract: The aim of this study was to assess the feasibility of detecting anti-HLA antibodies in eluates from needle core biopsies of renal transplants with chronic allograft nephropathy. Two methods of screening, the enzyme-linked immunosorbent assay (ELISA) and flow cytometry (FlowPRA) were compared. Twenty renal transplants with CAN were removed after irreversible graft failure. To assess the feasibility of detecting anti-HLA antibodies in small samples, needle core biopsies were sampled at the same place as surgical samples and at a second cortical area. Antibodies were eluted with an acid elution kit and anti-class I and class II IgG HLA antibodies detected using ELISA and flow cytometry. Flow cytometry was found to be more sensitive than ELISA for detecting anti-HLA antibodies in eluates from renal transplants with CAN (95% vs. 75% of positive cases). Detection of anti-HLA antibodies showed good agreement between surgical samples and needle core biopsies performed at the same place for anti-class I (80% vs. 65%, r=0.724 P<0.01) and anti-class II HLA antibodies (70% vs. 55%, r=0.827 P<0.01). In addition, differences in the detection of anti-class I HLA antibodies in needle core biopsies sampled at different sites suggests that immunization to class I donor antigen could be underestimated in needle core biopsy samples. These data indicate that anti-HLA antibodies can be detected in needle core biopsies from renal transplants. Provided further evaluation is done, elution might be a complementary method to detect anti-HLA antibodies when they are bound to the transplant.
authors.affiliation: Department of Pathology, School of Medicine, Dijon, France. laurent.martin@u-bourgogne.fr
authors.forename: Laurent
authors.initials: L
authors.lastname: Martin
authors.forename: Fredy
authors.initials: F
authors.lastname: Guignier
authors.forename: Olivier
authors.initials: O
authors.lastname: Bocrie
authors.forename: Philippe
authors.initials: P
authors.lastname: D'Athis
authors.forename: David
authors.initials: D
authors.lastname: Rageot
authors.forename: Gérard
authors.initials: G
authors.lastname: Rifle
authors.forename: Eve
authors.initials: E
authors.lastname: Justrabo
authors.forename: Christiane
authors.initials: C
authors.lastname: Mousson
mesh_terms: D000906:Antibodies / Q000032:analysis*; D001707:Biopsy, Needle; D002908:Chronic Disease; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D005240:Feasibility Studies; D005434:Flow Cytometry / Q000592:standards*; D006680:HLA Antigens / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D007668:Kidney / Q000276:immunology* / Q000473:pathology; D007674:Kidney Diseases / Q000209:etiology* / Q000276:immunology*; D016030:Kidney Transplantation / Q000009:adverse effects*; D012680:Sensitivity and Specificity; D014184:Transplantation, Homologous
chemical_list: D000906:Antibodies; D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II