title: Clinical cytometry and progress in HLA antibody detection.
abstract: For most solid organ and selected stem cell transplants, antibodies against mismatched HLA antigens can lead to early and late graft failure. In recognition of the clinical significance of these antibodies, HLA antibody identification is one of the most critical functions of histocompatibility laboratories. Early methods employed cumbersome and insensitive complement-dependent cytotoxicity assays with a visual read-out. A little over 20 years ago flow cytometry entered the realm of antibody detection with the introduction of the flow cytometric crossmatch. Cytometry's increased sensitivity and objectivity quickly earned it popularity as a preferred crossmatch method especially for sensitized recipients. Although a sensitive method, the flow crossmatch was criticized as being "too sensitive" as false positive reactions were a know drawback. In part, the shortcomings of the flow crossmatch were due to the lack of corresponding sensitive and specific HLA antibody screening assays. However, in the mid 1990s, solid phase assays, capable of utilizing standard flow cytometers, were developed. These assays used microparticles coated with purified HLA molecules. Hence, the era of solid-phase, microparticle technology for HLA antibody detection was born permitting the sensitive and specific detection of HLA antibody. It was now possible to provide better correlation between HLA antibody detection and the flow cytometric crossmatch. This flow-based technology was soon followed by adaptation to the Luminex platform permitting a mutltiplexed approach for the identification and characterization of HLA antibodies. It is hoped that these technologies will ultimately lead to the identification of parameters that best correlate with and/or predict transplant outcomes.
authors: Department of Pathology, Emory University, Atlanta, Georgia, USA.
authors: Robert A
authors: RA
authors: Bray
authors: Christine
authors: C
authors: Tarsitani
authors: Howard M
authors: HM
authors: Gebel
authors: Jar-How
authors: JH
authors: Lee
mesh_terms: D000906:Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology; D003165:Complement System Proteins / Q000737:chemistry / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods; D005434:Flow Cytometry / Q000379:methods*; D006084:Graft Rejection / Q000276:immunology / Q000517:prevention & control*; D006680:HLA Antigens / Q000737:chemistry* / Q000276:immunology; D006648:Histocompatibility / Q000276:immunology*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007118:Immunoassay; D008214:Lymphocytes / Q000737:chemistry / Q000166:cytology* / Q000276:immunology; D016377:Organ Transplantation; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000906:Antibodies; D006680:HLA Antigens; D011994:Recombinant Proteins; D003165:Complement System Proteins