title: Relatively high levels of anti-HLA antibodies (measured by Luminex Single-Antigen bead assay) are required to mediate inhibition of lymphocyte proliferation induced by sera from alloimmunized renal patients.
abstract: BACKGROUND
  Rejection of transplanted organs is caused by alloimmune responses, primarily against HLA molecules. Anti-donor HLA antibodies are associated with antibody-mediated rejection (AMR) and poor graft outcome. Because of clinical interest in detecting these antibodies, new technologies have recently been introduced to increase the sensitivity of detection. The Luminex Single-Antigen (LSA) bead assay may yield new information, but it must be validated against biological and clinical data.
  
  
  METHODS
  Based on previously published data regarding the in vitro effects of anti-HLA antibodies on lymphocytes, we measured the effect on lymphocytes of sera from patients on the transplant waiting list who had high titers of anti-HLA antibodies. Anti-CD3-mediated lymphocyte activation was studied in the presence of whole serum from these patients. Changes in lymphocyte proliferation, measured by carboxyfluorescein succinimidyl ester (CFSE) labeling, were detected, and these changes correlated with the level of anti-HLA antibodies.
  
  
  RESULTS
  Whole serum containing anti-HLA antibodies inhibited lymphocyte proliferation; this effect correlated with the level of antibodies, as measured by LSA. This inhibitory effect was HLA-specific, as shown by adsorption experiments. We also found that relatively high levels of anti-HLA antibodies were necessary to induce changes in an in vitro model of lymphocyte proliferation.
  
  
  CONCLUSIONS
  Our results demonstrate the clinical utility of detecting anti-HLA antibodies by LSA.
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Juan José
authors.initials: JJ
authors.lastname: Mata Molanes
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Leire
authors.initials: L
authors.lastname: Burgos Rodríguez
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Delgado García
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Pérez-Robles
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Moreno Narro
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Paula
authors.initials: P
authors.lastname: Arana Berganza
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Silvia
authors.initials: S
authors.lastname: Chocarro de Miguel
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Juana
authors.initials: J
authors.lastname: Merino Roncal
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Moreno Parado
authors.affiliation: Department of Immunology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Sánchez-Ibarrola
mesh_terms: D049109:Cell Proliferation; D006084:Graft Rejection / Q000276:immunology*; D006680:HLA Antigens / Q000276:immunology*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007518:Isoantibodies / Q000276:immunology*; D016030:Kidney Transplantation; D008213:Lymphocyte Activation / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology* / Q000502:physiology
chemical_list: D006680:HLA Antigens; D007518:Isoantibodies