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

Journal Article · Tue Nov 01 04:00:00 EST 1994 · Radiation Research

DOI:https://doi.org/10.2307/3578911· OSTI ID:70211

Sandru, G; Greiner, R ^[1]

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1. Univ. of Bern (Switzerland)

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

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
56 BIOLOGY AND MEDICINE
APPLIED STUDIES
ACTINOMYCIN
BIOLOGICAL REPAIR
BLOOD CELLS
CELL CULTURES
CONCANAVALIN A
DYNAMIC FUNCTION STUDIES
HEPARIN
INDUCTION
ION EXCHANGE CHROMATOGRAPHY
IRRADIATION
PHYTOHEMAGGLUTININ
PURIFICATION
PUROMYCIN
RADIATION INJURIES
SECRETION
THYMIDINE