Establishment of a human T-cell hybridoma that produces human macrophage activating factor for superoxide production and translation of messenger RNA of the factor in Xenopus laevis oocyte.
Human monoblast-like histiocytic lymphoma cell line U937 was induced by a macrophage activating factor for O2- production (MAF-O) to produce O2- in response to phorbol myristate acetate stimulation. A MAF-O-producing human T-cell hybridoma, F4-29-4, was established which was also found to produce macrophage activating factor for glucose consumption (MAF-G) and colony stimulating factor (CSF) when assayed against mouse bone marrow cells. MAF-O could be successfully separated from CSF but not from MAF-G by phenyl-Sepharose CL-4B chromatography (Phenyl-EG-fraction). To differentiate MAF-O from MAF-G and to explore a route for large scale production of MAF-O and its structural elucidation, total messenger RNA was extracted from a human T-cell hybridoma, clone F4-29-4. This messenger RNA was fractionated on 5-30% sucrose gradient and each fraction was microinjected into Xenopus laevis oocytes. MAF-O activity was found in the supernatant of oocytes injected with messenger RNA sedimentated at about 13.0S, while MAF-G messenger RNA was found to be about 10.5S. The MAF-O activity, synthesized from the injected messenger RNA, was not neutralized with an excess amount of anti-human IFN-gamma anti-serum, suggesting that MAF-O is antigenically different from human IFN-gamma.
D000818:Animals; D002847:Chromatography, Agarose; D003115:Colony-Stimulating Factors / Q000096:biosynthesis; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007371:Interferon-gamma / Q000276:immunology; D007700:Kinetics; D008222:Lymphokines / Q000096:biosynthesis*; D008262:Macrophage Activation; D016215:Macrophage-Activating Factors; D009865:Oocytes / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D013481:Superoxides / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology; D014982:Xenopus laevis
D003115:Colony-Stimulating Factors; D008222:Lymphokines; D016215:Macrophage-Activating Factors; D012333:RNA, Messenger; D013481:Superoxides; D007371:Interferon-gamma