Rapid assessment of the antigenic integrity of tetrameric HLA complexes by human monoclonal HLA antibodies.
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.
Eijsink
Chantal
C
Leiden University Medical Center, Department of Immunohaematology and Bloodtransfusion, Leiden, The Netherlands.
Kester
Michel G D
MG
Franke
Marry E I
ME
Franken
Kees L M C
KL
Heemskerk
Mirjam H M
MH
Claas
Frans H J
FH
Mulder
Arend
A
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
D000911:Antibodies, Monoclonal; D015234:HLA-A Antigens; D015395:Histocompatibility Antigens Class I; D011137:Polystyrenes