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Further Aspects of Microgranulocytotoxicity
Author(s) -
Drew S. I.,
Carter B. M.,
Guidera D.,
Lee K. E.,
Sasaki M.,
Terasaki P. I.,
Gale R. P.
Publication year - 1979
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1046/j.1537-2995.1979.19479250181.x
Subject(s) - cytotoxicity , granulocyte , serology , antibody , ficoll , immunology , in vitro , incubation , chemistry , cytotoxic t cell , lysis , sensitization , hemolysis , antigen , immune system , papain , microbiology and biotechnology , biology , biochemistry , enzyme , peripheral blood mononuclear cell
Some aspects influencing the serologic outcome of complement‐dependent granulocyte cytotoxicity are presented. Ficoll‐Hypaque (density 1.060) gradient centrif‐ugation with hypotonic lysis of red blood cells yielded populations of granulocytes with greater than 90 per cent purity. Granulocytes exposed to papain (0.01%) at 24 C for 12 minutes showed enhanced serologic reactivity compared with untreated cells. In addition, enhanced cytotoxicity was promoted by 5 C (cold) as opposed to 22 C (warm) incubation of granulocytes and antibody in the first stage of the microcytotoxicity assay and emphasizes the temperature dependence of the serologic reactions. Neutrophil‐specific leukoagglutinins were not cytotoxic under optimal in vitro conditions. Conversely, a significant proportion of antibodies selected for cytotoxicity failed to agglutinate granulocytes. Cold reactive granulo‐cytotoxins are complement dependent, IgM in nature, and 2‐mercaptoethanol sensitive and do not appear to be immune complexes. By optimizing reaction conditions, granulocyte microcytotoxicity is a valuable addition to in vitro assays detecting immune sensitization against granulocyte surface antigens.