pactamycin and Mast-Cell-Sarcoma

Target cells (P-815 mastocytoma cells) infected with ectromelia virus became susceptible to lysis by H-2 compatible specific effector T cells within one hour of exposure of the cells to virus. This is long before viral progeny are produced and shed from the cell. Polyacrylamide gel electrophoresis (PAGE) profiles of the plasma membranes from infected and uninfected P-815 cells pulsed with 35S-methionine for one or a few hours after infection with virus were very complex and showed no consistent differences. P-815 cells, infected with ectromelia virus in the presence of an inhibitor of protein synthesis, pactamycin, slowly became susceptible to cell mediate lysis when the pactamycin was removed. The number of polypeptide species synthesized under these conditions was reduced to only three, of molecular weights between 10,000-50,000 daltons. Specific, newly synthesized membrane components recognized by mouse convalescent sera were isolated by immune complexing and examined by PAGE. Six polypeptide bands were seen, the major one correlating with one observed in the pactamycin experiment. The results suggested that the convalescent serum recognized both viral and host cell coded antigens. The significance of these findings is discussed.

Topics: Animals; Antibodies, Viral; Antigens; Cell Line; Cell Membrane; Cytotoxicity Tests, Immunologic; Depression, Chemical; Ectromelia virus; Ectromelia, Infectious; Mast-Cell Sarcoma; Membrane Proteins; Methionine; Mice; Pactamycin; T-Lymphocytes