title: The single antigen expressing lines (SALs) concept: an excellent tool for screening for HLA-specific antibodies.
abstract: Definition of the antibody specificity in the serum of patients waiting for a renal transplant or in need for platelet transfusion is a crucial step for finding adequate donors. Confounding factors are the complexity of the serum antibodies and the expression of several, up to six, different human leukocyte antigens (HLA) on peripheral blood lymphocytes used as target cells in the antibody screening. Single antigen-expressing (SAL) cell lines were generated by transfecting human major histocompatibility complex (MHC) class I sequences into K562, an erythroleukemia-derived cell line lacking MHC class I and II expression. Thirty-seven different SALs have been generated so far. In this study, we present the validation of 16 of those SALs by flow cytometry against a panel of 84 human HLA-specific monoclonal antibodies (30 HLA-A [8 IgG/22 IgM], 45 HLA-B [18 IgG/27 IgM], 6 HLA-A, B [3 IgG/3 IgM], and 3 HLA-C [all IgM]) developed in our laboratory. The SALs proved to be suitable tools to determine acceptable mismatches for highly sensitized patients. This concept of transfecting target sequences in immortalized cell lines opens up new avenues in the definition of serum and cellular reactivity for sensitized patients awaiting a suitable organ or blood component.
authors.affiliation: Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Yvonne M
authors.initials: YM
authors.lastname: Zoet
authors.forename: Chantal
authors.initials: C
authors.lastname: Eijsink
authors.forename: Marrie J
authors.initials: MJ
authors.lastname: Kardol
authors.forename: Marry E I
authors.initials: ME
authors.lastname: Franke-van Dijk
authors.forename: G Louis
authors.initials: GL
authors.lastname: Wilson
authors.forename: Roel
authors.initials: R
authors.lastname: de Paus
authors.forename: Eric
authors.initials: E
authors.lastname: Mickelson
authors.forename: Mirjam
authors.initials: M
authors.lastname: Heemskerk
authors.forename: Peter J
authors.initials: PJ
authors.lastname: van den Elsen
authors.forename: Frans H J
authors.initials: FH
authors.lastname: Claas
authors.forename: Arend
authors.initials: A
authors.lastname: Mulder
authors.forename: Ilias I N
authors.initials: II
authors.lastname: Doxiadis
mesh_terms: D000906:Antibodies / Q000097:blood / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D045744:Cell Line, Tumor; D003165:Complement System Proteins / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic; D005260:Female; D005434:Flow Cytometry; D006680:HLA Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology; D015791:HLA-A3 Antigen / Q000235:genetics / Q000276:immunology; D015793:HLA-B7 Antigen / Q000235:genetics / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D020014:K562 Cells; D007963:Leukocytes, Mononuclear / Q000276:immunology; D014162:Transfection
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D006680:HLA Antigens; D015789:HLA-A2 Antigen; D015791:HLA-A3 Antigen; D015793:HLA-B7 Antigen; D015395:Histocompatibility Antigens Class I; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D003165:Complement System Proteins