Detection of Leukocyte Antibodies by Means of I131-Labelled, Purified Anti—Human-Globulin Antibody. Problems of Nonspecific Adsorption of Globulin by Leukocytes

Abstract

Abstract Initial studies of a quantitative method for detecting sensitization of white blood cells by antileukocyte antibodies are presented. The method entails isolating I131-labelled, anti—human-globulin antibody (prepared in rabbits) by adsorption and elution from insoluble antigen, globulinazobenzylcellulose. The degree of adsorption of the labelled antibody by presumably sensitized as opposed to nonsensitized leukocytes is measured by radioactive isotope counting technics. The vexing problem of nonspecific adsorption of human or rabbit globulin by leukocytes is considered, and procedures for partially circumventing this problem are set forth. Seligmann's solution with normal rabbit serum added to a final concentration of 10 per cent appeared to be the most effective washing fluid in most instances for removing nonspecifically adsorbed globulins from leukocytes. The use of purified anti—human-globulin antibody, as opposed to whole Coombs serum, also very favorably affected the ratio between immunologically specific adsorption and nonspecific adsorption. Results of the over-all I131 method as applied to selected positive and negative human sera are detailed.