Blood, 1960, Vol. 16, No. 5, pp. 1523-1534.
© 1960 American Society of Hematology, Inc.
Detection of Leukocyte Antibodies by Means of I131-Labelled,
Purified Anti—Human-Globulin Antibody. Problems of
Nonspecific Adsorption of Globulin by Leukocytes
ROBERT E. ANDERSON 1 and
ROY L. WALFORD 1
1 Department of Pathology, University of California School of Medicine, Los
Angeles, Calif.
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.
Submitted on April 28, 1960
Accepted on July 3, 1960
