EFFECT OF ANTIBODY ON HETEROLOGOUS AND HOMOLOGOUS CELLS IN TISSUE CULTURE

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  • 1 January 1963
    • journal article
    • research article
    • Vol. 6  (6) , 569-+
Abstract
In order to compare antibody against heterologous and homologous tissue in the same serum, rabbits were immunized with rat kidney and complete adjuvant. The resulting sera showed antibody against both rat and rabbit kidney. Cultures of rat kidney cells were killed by exposure to these sera. A concentration of 0.3 per cent gamma-globulin (ammonium sulphate fraction) was adequate to kill cultures in the absence of complement, but smaller concentrations were effective when guinea-pig complement was added. The cell surfaces were shown to have taken up antibody by the fluorescent antibody technique. Cytoplasmic staining could only be shown in cells which had previously been injured by freezing and thawing or by fixation. Rabbit kidney cells in culture were unaffected when exposed to whole rabbit anti-rat serum, but were killed and their cell membranes stained on exposure to gamma-globulin derived by (NH4)2SO4-fractionation from such serum and having the same complement-fixation titre as the parent serum. At least 0.6 per cent gamma-globulin had to be added to kill rabbit cell cultures. It was found that normal rabbit serum had a partial protective effect against this antibody. Fractionation of sera by gradient centrifugation or chromatography or DEAE-cellulose showed that while antibody against heterologous tissue was found both in the 7S gamma-globulin and macroglobulin fractions, antibody against homologous tissue was confined to the latter. It is considered that the findings do not support a concept of an in vivo pathogenic role for circulating antibody.