Serum-mediated modification of proliferation in factor-dependent macrophage cell lines.
Previously we described that bacterial lipopolysaccharide (LPS) promoted DNA synthesis and supported the cell viability in the factor-dependent macrophage cell lines BDM-1 and BDM-1W3 in the absence of colony-stimulating factor (CSF). To further examine this phenomenon, in the present study we examined the effects of serum on CSF-dependent proliferation and LPS-induced DNA synthesis in BDM-1 and BDM-1W3 cells. Fetal calf serum (FCS) was required for CSF-dependent proliferation in BDM-1 and BDM-1W3 cells. FCS was also required for LPS-induced DNA synthesis in BDM-1W3 cells. However, at concentrations higher than 0.2%, FCS inhibited LPS-induced DNA synthesis in BDM-1W3 cells in a dose-dependent manner. To obtain the inhibitory activity in FCS (FCS-In) for LPS-induced DNA synthesis, FCS was fractionated by gel filtration chromatography using Sephacryl S-200, chromatography on DEAE-Sephacel, and affinity chromatography on heparin-Sepharose. FCS-In was eluted in the void volume peak from a Sephacryl S-200 column, indicating that FCS-In has a molecular weight of more than 250,000. The molecular weight of FCS-In was apparently 270,000 as determined by SDS-polyacrylamide gel electrophoresis (PAGE) under non-reducing conditions. Upon reduction, four components became detectable with apparent molecular weights of 170,000, 110,000, 67,000, and 30,000. The inhibitory activity in FCS-In material was inactivated by heat and trypsin treatment. The partially purified FCS-In inhibited LPS-induced DNA synthesis in BDM-1W3 cells, but did not inhibit the proliferation of BDM-1W3 cells induced by IL-3, granulocyte-macrophage CSF (GM-CSF), or macrophage CSF (M-CSF).(ABSTRACT TRUNCATED AT 250 WORDS)
Ohki
K
K
Department of Microbiology, Saga Medical School, Japan.
Kohashi
O
O
D000818:Animals; D001769:Blood; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D002555:Cerebrospinal Fluid / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid; D003470:Culture Media; D004247:DNA / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D008970:Molecular Weight
D003470:Culture Media; D008070:Lipopolysaccharides; D004247:DNA