Acid treatment of lymphocytes selectively decreases the expression of HLA class I antigens: a method to confirm that a positive clinical crossmatch test was due to class I antibodies.
A rapid and reliable method for eliminating HLA class I antigens from the surface of lymphocytes without damaging the cells is described. Lymphocytes were exposed to an acid solution (pH 3.0) which selectively destroys the antigenicity of HLA class I antigens. Alloantisera containing multispecific HLA class I antibodies reacted with phosphate-buffered saline (PBS)-treated, but not with acid-treated, lymphocytes. Specificity controls included: antibodies against HLA class II antigens and CD3, CD4, CD8; markers expressed on T cells and CD19, CD23; markers expressed on B cells. No change in lymphocyte reactivity to any of these surface antigens or to autoantibodies was observed. The viability of acid-treated lymphocytes was regularly around 90%. We propose that acid-treated lymphocytes are suitable targets for determination of the sole presence of class I antibodies in crossmatch sera of patients awaiting organ transplants.
Sumitran-Karuppan
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S
Division of Immunology, Microbiology, Pathology and Infectious Diseases F-79, Karolinska Institute, Huddinge Hospital, Sweden.
Möller
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D000143:Acids / Q000494:pharmacology*; D019343:Citric Acid / Q000494:pharmacology; D015536:Down-Regulation / Q000276:immunology*; D006680:HLA Antigens / Q000096:biosynthesis / Q000187:drug effects* / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis / Q000187:drug effects* / Q000276:immunology*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007518:Isoantibodies / Q000276:immunology*; D010710:Phosphates / Q000494:pharmacology
D000143:Acids; D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D007518:Isoantibodies; D010710:Phosphates; D019343:Citric Acid