| Project/Area Number |
02670152
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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 |
Experimental pathology
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| Research Institution | YAMAGUCHI UNIVERSITY |
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Principal Investigator |
UCHINO Fumiya (1991-1992) YAMAGUCHI UNIV. SCH. OF MED. PROFESSOR, 医学部, 教授 (20034902)
横田 忠明 (1990) 山口大学, 医学部, 講師 (40144927)
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| Co-Investigator(Kenkyū-buntansha) |
KAWANO Hiroo YAMAGUCHI UNIV. SCH. OF MED. RESEARCH ASSOCIATE, 医学部, 助手 (40234091)
ISHIHARA Tokuhiro YAMAGUCHI UNIV. SCH. OF MED. ASSOCIATE PROF., 医学部, 助教授 (70089910)
内野 文彌 山口大学, 医学部, 教授 (20034902)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1992: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1990: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Amyloid enhancing Factor / Ubiquitin / Experimental Amyloidosis / Neutrophil / Immunoelectron Microscopy / Ubiquitin, / アミロイド-シス / AEF / アミロイド / アミロイドーシス / amyloid enhancing factor / 好中球 |
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| Research Abstract |
Amyloid enhancing factor (AEF) is one of the factors in amyloidogenesis, besides the increase of amyloid precursor protein in a serum.
We present the following results;
1. We extracted AEF from organs of amyloidotic mice and the highest AEF activity was found in a spleen. On a histochemical study using anti-AEF antiserum, derived from the crude AEF, AEF is localized on neutrophils of spleen and peripheral blood.
Electron microscopy reveals that AEF is not distributed in granules but in the cytosol of a neutrophil. Furthermore collected neutrophils are separated into granular and extra-granular fractions, with adding a degranulating agent, and only the granular fraction has AEF activity on a trial study on mice.
2. AEF is purified with a DEAE ion exchange column and a reverse phase HPLC from neutrophils of spleen and peripheral blood treated with amyloidotic stimuli. AEF activity is in a fraction eluted with 0.17M saline on DEAE column and in a peak of retention time 23 min. on HPLC. SDS-page gives single band, about 15 KDa. considering with a result from a Gas-chromatography AEF is a glycoprotein weighing 15 KDa.
3. Kupffer cells, cultured in the media with AEF, have many cell processes and swell, and on electron microscopic observation they indicate degenerations of organelle.
These results suggest that AEF may cause damage on macrophages and then amyloid precursor protein increased with amyloidotic stimuli, which is not able to be degradated, is polymerized and changes into fibrils.
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