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Enhancement in irradiated mononuclear cells in culture of mitogen-induced incorporation of [{sup 3}H]thymidine by homologous conditioned medium

Journal Article · · Radiation Research
DOI:https://doi.org/10.2307/3578911· OSTI ID:70211

Incorporation of [{sup 3}H]thymidine in irradiated peripheral blood mononuclear cell cultures irradiated in vitro was stimulated significantly by either concanavalin A or phytohemagglutinin only in the presence of homologous conditioned medium. Production of this activity by mononuclear cells was enhanced by irradiation and/or pulsed exposure to puromycin but was abolished by actinomycin D. Addition of anti-interleukin 1 or anti-interleukin 2 monoclonal antibodies to the conditioned medium before assay did not influence the stimulatory action. A similar significant stimulation of mononuclear cell cultures irradiated with 6 Gy by concanavalin A was obtained when purified preparations of homologous conditioned medium were used in the assay. Purification was done by ultrafiltration and concentration, heparin agarose chromatography, ammonium sulfate precipitation, concanavalin A agarose chromatography, DEAE-ion exchange chromatography and HPLC gel filtration chromatography. With SDS-PAGE and silver staining, the active HPLC fraction gave one band of 50 kDa, suggesting that this protein is responsible for the co-stimulatory effect of homologous conditioned medium for both mitogen-induced irradiated and nonirradiated mononuclear cell cultures. 42 refs., 9 figs., 3 tabs.

Sponsoring Organization:
USDOE
OSTI ID:
70211
Journal Information:
Radiation Research, Journal Name: Radiation Research Journal Issue: 2 Vol. 140; ISSN RAREAE; ISSN 0033-7587
Country of Publication:
United States
Language:
English