Purification of mouse cell interferon induced by isotopically labelled double-stranded f2 phage RNA.
The dynamics of interferon formation by an established cell line of mouse fibroblast (L cells) and by mouse peritoneal leukocytes induced by double-stranded RNA extracted from E. coli f2 phage is described. The L cells produced interferon at a lower rate, the maximum values were obtained at 12 to 20 hours after induction, and the production was ultimately dependent on the established cell line used and on the presence of DEAE-dextran during induction. The mouse peritoneal leukocytes (MPL), on the other hand, did not require DEAE-dextran and the maximum of interferon production was reached between 6 and 12 hours after induction. Both the L cell- and the MPL-interferons were purified and concentrated so that the final specific biologic activity was 100-to 300-fold higher than that of the initial preparations (1 to 5 X 10(6) interferon units per mg protein). Polyacrylamide gel electrophoresis showed similar migration profiles for the preparations of both interferons. The smaller part of the activity was situated in a broader, slow-moving peak and the greater part formed a sharp, high and fast-moving peak. Using 3H uridine-labelled f2 ds-RNA for induction of interferon it was found that one of the radioactivity zones coincided with the fast-moving activity peak of the purified and concentrated interferon.
D000818:Animals; D002850:Chromatography, Gel; D003090:Coliphages; D004591:Electrophoresis, Polyacrylamide Gel; D007369:Interferon Inducers; D007372:Interferons / Q000096:biosynthesis / Q000302:isolation & purification*; D007739:L Cells / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism; D051379:Mice; D010761:Phosphorus Radioisotopes; D012367:RNA, Viral / Q000494:pharmacology*; D014529:Uridine / Q000378:metabolism
D007369:Interferon Inducers; D010761:Phosphorus Radioisotopes; D012367:RNA, Viral; D007372:Interferons; D014529:Uridine