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  • Title: Isolation and characterization of IFN-gamma mRNA derived from mitogen-induced human splenocytes.
    Author: Devos R, Cheroutre H, Taya Y, Fiers W.
    Journal: J Interferon Res; 1982; 2(3):409-20. PubMed ID: 6182256.
    Abstract:
    Human splenocytes from large cultures (5 1) were stimulated with Staphylococcus enterotoxin A (SEA). After 2.5 d of conditioning, poly A+RNA was isolated from these cultures and its in-vitro translational capacity was evaluated in a wheat germ extract. Injection of the RNA into Xenopus laevis oocytes yielded a low but reproducible amount of IFN activity, which was neutralizable only by an antiserum to gamma interferon (IFN-gamma) and not by antisera to alpha interferon (IFN-alpha) and/or beta interferon (IFN-beta). Formamide sucrose gradient centrifugation analysis indicated that the mRNA which codes for IFN-gamma sediments at around 15 S. The profile, however, suggested a size heterogeneity of IFN-specific mRNA. Electrophoresis of cDNA synthesized on these mRNA fractions indicated that the mRNA coding for gamma IFN has a length of between 900 and 1400 nucleotides.
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