Prification of an Antitumor Factor Induced in Hepatocytes by Interferon and cDNA cloning of the factor
| Project/Area Number |
04680178
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Osaka Bioscience Institute |
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Principal Investigator |
YASUI Hiroaki Researcher, Departiment of Cell Biology, Osaka Bioscience Institute, 第4研究部, 研究員 (80230209)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIDA Ryotaro Head, Department of Cell Biology, Osaka Bioscience Institute, 第4研究部, 部長 (10124760)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Interferon / Hepatocyte / Tumor cell / インターフェロン / 肝細胞 / Hepatocytes(肝細胞) |
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| Research Abstract |
We have reported previously that IFn-alpha/beta-reated fepatocytes in culture release a soluble factor(s) which suppress the multiplication of an IFN-alpha/beta-resistant clone of Friend leukemia cells (FLCs). To characterize the factor(s) further, we first examined the possibility that products of non-parenchymal cells (NPCs) included in small number in the hepatocyte cultures were involved in the inhibitory activity. We prepared cultures of purified adherent NPCs, mostly Kupffer cells and sinusoidal endothelial cells, and culture supernatants of NPCs pretreated with IFN-alpha/beta were tested for the inhibitory activity for FLC multiplication. IFN did not induce any inhibitory activity in NPC cultures, while LPS-stimulated NPCs cultivated in parallel released several inhibitory factors inculding TNF-alpha. To explore the possibility that IFN augmented the release of hepatocyte cytosolic proteins including arginase, we compared the inhibitory activity in culture supernatant of IFN-treated hepatocytes with that found in hepatocyte extract by anion exchange chromatography. The IFN-induced inhibitory activity was eluted at relatively high salt concentration as a single peak, while the inhibitory activity in hepatocyte extract was co-eluted with arginase at low salt concentration. To purify the factor further, eluates containing the inhibitory activity were concentrated, and the concentrate was subjected to gel filtration chromatography. The inhibitory activity specifically induced by IFN was eluted at around 40 KDa as a single peak. Thus, partial purification of the inhibitor specifically induced by IFN-alpha/beta in hepatocyte cultures was achieved.
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Report
(3 results)
Research Products
(5 results)
