Cytotoxic Effects of Antisera Against Huma Epithelial Cells Grown in Tissue Culture

Studies on Immune Cellular Injury

“…The cytotoxicity of human serum could not be demonstrated following heating for 30 minutes at 56 ° or in the absence of antibody. These qualitative experiments therefore confirmed and extended previous observations(2)(3)(4)(5)(6)(7)(9)(10)(11)(12)(13) and provided the basis for subsequent experiments on the nature of the normal, heat-labile, human serum factors required for immune cytotoxicity.…”

“…In certain previous studies in which both antibody and normal serum were required for cytotoxicity, identification of the normal serum constituents with complement was made primarily on the basis of heat-lability (56 ° , 30 minutes) (2)(3)(4)(5)(6)(7)(9)(10)(11)(12)(13). One report showed further that complement-fixed normal guinea pig serum, as well as heated normal guinea pig serum, was without cytotoxic activity on chick embryo hearts sensitized with specific guinea pig antibody (42).…”

“…In this connection, it is of interest that Lumsden (39), Bolande and Todd (41), and others (13,40) have found that normal serums, in the absence of demonstrable antibody, have cytotoxic activity for certain established cell lines in tissue culture. On the other hand, numerous investigators have demonstrated the necessity for specific cytotoxic antibodies (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(36)(37)(38)(39)(40). Ross has delineated five criteria for establishing the participation of specific anticeUular antibodies in cytotoxicity (43).…”

“…The conclusions drawn from the data presented have been based in most instances on two independent criteria of cytotoxicity: metabolism-inhibition and uptake of trypan blue. Both of these techniques have been widely used as indicators of cell viability under well defined experimental conditions (2,5,6,8,(14)(15)(16)(44)(45)(46). Uptake of trypan blue proved to be a rapid, convenient, and reproducible procedure for measuring the cytotoxic activity of normal human serum on human amnion cells sensitized with rabbit antibody.…”

Cultural Characterization of Animal Cells

“…Immunologic characterization of cells.-Although immunologic reac tions of cells have been measured by various forms of the cytotoxicity test (3,96,122,134,198), direct (37,123 ) and indirect (115, 23 1) hemaggluti nation, mixed-cell agglutination (62), complement-fixation (186,313), macroscopic agglutination (185 ), and metabolism inhibition (228,325), these techniques are limited by the requirement for large numbers of cells, dispersed cells, or fract i onation of cells for antigens. Until genetically pure cultures can be obtained readily, it is desirable to characterize cells either in colonies or from single colonies.…”