title: Evaluation of a new flow cytometry crossmatch procedure for simultaneous detection of cytotoxicity and antibody binding.
abstract: In this study we have evaluated an alternative 96-well format flow cytometry based (FCtox) method which enable simultaneous detection of cytotoxicity and human leukocyte antigen (HLA) antibody binding. Comparable results were obtained in side-by-side comparisons with conventional complement-dependent cytotoxicity (CDC) and flow cytometric crossmatch (FCXM) in terms of sensitivity and specificity. There was 91 and 93% agreement between results obtained by FCtox and CDC for T and B cells, respectively. In addition, comparable results were obtained with FCtox IgG and FCXM IgG for both T and B cells. Furthermore, compared with a recently developed and highly sensitive Luminex based C1q assay we obtained close to 90% method agreement with the FCtox assay. Our alternative cytotoxicity and IgG binding assay which exhibit low intra-and inter-assay variation will improve the workflow and speed up the pre-transplant testing and also allow continuous monitoring of assay performance and proper quality assurance.
authors.affiliation: Department of Laboratory Medicine, Division of Clinical Immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden. mats.alheim@karolinska.se
authors.forename: M
authors.initials: M
authors.lastname: Alheim
authors.forename: P K
authors.initials: PK
authors.lastname: Paul
authors.forename: D-M
authors.initials: DM
authors.lastname: Hauzenberger
authors.forename: A-C
authors.initials: AC
authors.lastname: Wikström
mesh_terms: D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods* / Q000592:standards; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D008297:Male; D015588:Observer Variation; D011485:Protein Binding; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology
chemical_list: D007074:Immunoglobulin G
keywords: complement-dependent cytotoxicity; crossmatching; flow cytometry; human leukocyte antigen antibodies; kidney transplantation