Some cytochemical investigations were carried out on the effect of antisera produced in whole cells, free nuclei, and mitochondrial fraction of HeLa cells and upon HeLa cells in tissue culture, in the hope of gaining some critical information for understanding the mechanism of injurious effect upon tumor cells
in vitro.
The antisera affected the cells very actively in a dilution of 1:4 to 1:8 with nutrient medium in the presence of a heat-labile complement-like factor. The HeLa cells exposed to the antisera were damaged within 12 hours through the deformation, pycnosis, and fragmentation of nuclei, and the aggregation of cytoplasmic granules, together with the loss of cytoplasmic basophilia. The reaction rapidly became especially marked on the cell surface. After several cytochemical tests, it was found that there occured a disappearance of ribonucleic acid from the cytoplasm of the affected cells, along with a denaturation or condensation of some proteins.
Several enzymatic studies revealed that the HeLa cells treated with antiserum utilized exogenous succinate, but did not metabolize exogenous glucose, and that peroxidase granules made their appearance in the affected cells, this being the sign of cell damage.
The antisera were found to be antigen-cell specific, so far as the present experiments are concerned. They lost their cytotoxic activity by incubation of the serum at 56° for 30 minutes.
The whole cells, free nuclei, and mitochondrial fraction of HeLa cells were all found to be antigenic and almost equally elicited potent cytotoxic antibodies in heterologous hosts.
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