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Effects of Antibodies on L5178Y Mouse Leukemia Cells Cultured in Vitro¹

Department of Microbiology and Immunology, McGill University, Montreal 110, Quebec, Canada

An in vitro system with the use of a rabbit serum medium-adapted subline of L5178Y cells and rabbit antisera was used in the present study.

Heat-inactivated antisera inhibited both uptake and incorporation of glycine-2-¹⁴C into cell protein and killed the cells. These effects became demonstrable only after 6 hr and reached a maximum after 24 to 48 hr. Total cell counts decreased even more slowly and the initial cause of cell death did not seem to result from cell lysis.

Sodium succinate partially protected the cells against killing or inhibition by antisera. Viable cell counts, uptake, and incorporation of glycine-2-¹⁴C increased greatly. Malonate reversed completely the action of succinate. Apparently, the energy derived from the oxidation of succinate is sufficient for many cells to grow and incorporate the amino acid into cell protein. Glucose and nicotinamide also showed similar protective effects.

Antiserum was observed to have a transient stimulatory effect on viable cell counts, uptake, and incorporation of glycine-2-¹⁴C at about 2 hr after the addition of antisera. Whether this is due to stimulation of noncycling G2 cells by antiserum is currently under study.

During the entire experimental period, i.e., up to 4 days, a small portion of the cell population remained viable. Many of these looked much larger and the incorporation rate, but not uptake rate, of glycine-2-¹⁴C/10⁶ viable cells was much higher than that of the controls. Addition of succinate increased the rate still further. Cultures treated with antiserum recovered if fresh medium without antisera was added at 24 to 48 hr.

Possible modes of action of the antisera were discussed.

¹ This investigation was supported by a grant from the Medical Research Council of Canada.

² Holder of a Medical Research Council studentship.

Received 9/ 8/69. Accepted 10/30/69.