Interferon induced in human leukocytes by mitogens: production, partial purification and characterization.
Interferon was induced in leukocyte suspensions from human buffy coats by exposure to phytohemagglutinin P, concanavalin A (Con A) and staphylococcal enterotoxin A, under a variety of cell culture conditions. Con A was found to rapidly (within 12 h) induce high yields of antiviral activity (1.5 units/1000 cells). Lesser yields were obtained with the other two mitogens studied. The interferon was partially purified to a spec. act. around 10(5.3) units/mg protein, by batch adsorption on controlled-pore glass (CPG) beads and desorption by ethylene glycol. This material was characterized as containing mainly gamma-type interferon. Specifically on gel filtration, a fraction of 45000 daltons was obtained which could account for virtually all antiviral activity present in the starting material. Furthermore, the ethylene glycol-eluted antiviral activity was acid-labile, serologically distinct from alpha and beta-type interferon and strictly species-specific (no activity detectable on any of the available cell species sensitive to alpha and beta-type interferon). The crude culture supernatant also contained some antiviral activity which resembled beta-type interferon in that it adsorbed to CPG, could be desorbed by pH 2 buffer, was acid-resistant and could be neutralized by a specific anti-fibroblast interferon antiserum. The CPG/ethylene glycol-purified gamma-type interefron preparation was found to inhibit the growth of certain lymphoblastoid cells (Daudi and Molt-4). It also potentiated natural killer activity of fresh donor lymphocytes. In both respects, the gamma-type interferon preparation was not significantly more active than preparations of alpha and beta-interferon of similar antiviral potency.
D055598:Chemical Phenomena; D002621:Chemistry; D003208:Concanavalin A / Q000494:pharmacology; D005026:Ethylene Glycols; D005898:Glass; D006801:Humans; D007369:Interferon Inducers / Q000494:pharmacology*; D007372:Interferons / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D007962:Leukocytes; D008934:Mitogens / Q000494:pharmacology*
D005026:Ethylene Glycols; D007369:Interferon Inducers; D008934:Mitogens; D003208:Concanavalin A; D007372:Interferons