Tumoricidal effector mechanisms of murine BCG-activated macrophages. I. Parameters of production and initial characterization of a cytolytic factor serologically related to necrosin.
Previous studies have shown that peritoneal macrophages from mice chronically infected with Bacillus Calmette-Guérin (BCG) become highly cytotoxic for tumor targets upon further in vitro triggering with a variety of agents. In the current studies, achievement of this activated state was characterized by the production and release of a cytotoxin, herein termed cytolytic factor (CF), which appeared in the fluid phase. Production/release of CF by the macrophage required transcription, translation, glycosylation, and an intact secretory apparatus, as evident from inhibition by treatment with actinomycin-D, cycloheximide, tunicamycin, and monensin, respectively, prior to and during triggering with lipopolysaccharide (LPS). CF obtained by culture of BCG-activated macrophages appeared rapidly in the supernatant after triggering. Using the actinomycin-D-treated L-929 or EMT-6 targets in a microassay, CF secreted by macrophages cultured in low molecular weight serum components was detected as an approximately 150-kD component on Sephacryl S-200. CF demonstrated a spectrum of cytotoxic activity against a number of tumor and normal targets in vitro. Its lytic activity appeared equally effective whether the targets were cultured in medium containing fetal calf serum (FCS) or in Neuman-Tytell medium without serum during the assay. CF was moderately sensitive to treatment with TLCK and TAME; however, its activity in serum and apparent molecular weight distinguish it from a moiety obtained from BCG-activated murine macrophages and previously described. A rabbit heteroantiserum raised against highly purified necrosin, a product of the murine macrophage cell line J774.1, was extremely effective in neutralizing the biological activity of CF.
Klostergaard
J
J
Foster
W A
WA
Leroux
M E
ME
D000818:Animals; D001500:BCG Vaccine; D003429:Cross Reactions; D003602:Cytotoxicity, Immunologic; D003603:Cytotoxins / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism; D066298:In Vitro Techniques; D008264:Macrophages / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D008297:Male; D051379:Mice; D009374:Neoplasms, Experimental / Q000276:immunology*; D010447:Peptide Hydrolases / Q000378:metabolism; D011506:Proteins
D001500:BCG Vaccine; D003603:Cytotoxins; D011506:Proteins; D010447:Peptide Hydrolases