title: Rapid assessment of the antigenic integrity of tetrameric HLA complexes by human monoclonal HLA antibodies.
abstract: The ability of tetrameric major histocompatibility complex (MHC) class I-peptide complexes (tetramers) to detect antigen-specific T lymphocyte responses has yielded significant information about the generation of in vivo immunity in numerous antigenic systems. Here we present a novel method for rapid validation of tetrameric HLA molecules based on the presence of allodeterminants. Human monoclonal antibodies (mAbs) recognizing polymorphic determinants on HLA class I were immobilized on polystyrene microparticles and used to probe the structural integrity of tetrameric HLA class I molecules by flow cytometry. A total of 22 tetramers, based on HLA-A1, A2, A3, A24, B7 and B8 were reactive with their counterpart mAbs, thus confirming their antigenic integrity. A positive outcome of this mAb test ensures that tetrameric HLA class I can be used with greater confidence in subsequent functional assays.
authors.affiliation: Leiden University Medical Center, Department of Immunohaematology and Bloodtransfusion, Leiden, The Netherlands.
authors.forename: Chantal
authors.initials: C
authors.lastname: Eijsink
authors.forename: Michel G D
authors.initials: MG
authors.lastname: Kester
authors.forename: Marry E I
authors.initials: ME
authors.lastname: Franke
authors.forename: Kees L M C
authors.initials: KL
authors.lastname: Franken
authors.forename: Mirjam H M
authors.initials: MH
authors.lastname: Heemskerk
authors.forename: Frans H J
authors.initials: FH
authors.lastname: Claas
authors.forename: Arend
authors.initials: A
authors.lastname: Mulder
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D015234:HLA-A Antigens / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006358:Hot Temperature; D006801:Humans; D008863:Microspheres; D011110:Polymorphism, Genetic; D011137:Polystyrenes / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D015234:HLA-A Antigens; D015395:Histocompatibility Antigens Class I; D011137:Polystyrenes