| Project/Area Number |
06660395
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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 |
Applied veterinary science
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
FUJINAGA Toru Hokkaido Univ., Graduate School of Veterinary Medicine, Professor, 大学院獣医学研究科, 教授 (50181376)
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| Co-Investigator(Kenkyū-buntansha) |
OKUMURA Masahiro Hokkaido Univ., Graduate School of Veterinary Medicine, Instructor, 大学院獣医学研究科, 助手 (80260397)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Acute-phase protein / Acute-phas response / Cytokine / Dog / Horse / Sandwitch ELISA / Serum amyloid A protein / Slidereversed passive latex agguruyination test / インターロイキン-6 / 血清アルミロイドA蛋白 / α^1-酸性糖蛋 / インターロイキン-1 / 腫瘍壊死因子(TNF) / C-反応性蛋白 |
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| Research Abstract |
Bioassays for the measurement of inflammatory cytokines : interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF)-like activities in canine sera were conducted using human melanoma A375S1, IL-6 dependent murine hybridoma MG60・BSF2 and WEHI 164 murine sarcoma subclone 28-4 respectively. IL-1-like activity in sera from dogs with endotoxic shock. IL-6-and TNF-like activities were detected in sera from dogs with endotoxic shock and local inflammation.
The changes in serum activites of IL-1, IL-6 and TNF were studied in dogs with acute inflammation. Dogs with local inflammation induced by an intramuacular injection of turpentine oil showed a clinically typical progression in the inflammatory respnse and recovering on day 14 after the treatment. Serum concentration of acute-phase protein increased, while the albumin concentration decreased in dogs during the acute-phase response. Serum cytokine activities changed faster than acute-phase proteins and other reactans after local injury in gogs.
… More
Particulary, serum IL-6-like activity in creased dramatically, therefore, it may have a major role in the acute-phase response. Furthermore, the changes in serum cytokine activities induced by local injuries seems to be different from that induced by endotoxic shock.
The responses of inflammatory cytokines and acute-phase proteins in sera from horses with inflammatory conditions were basically the same as the changes in dogs.
For the measurement of the concentration of serum amyloid A (SAA) protein in horses, a sensitive and highly reproducible sandwick ELISA was established, using an affinity-purified SAA antibody. Results of the ELISA were found to have a high correlation (r=0.95) with those of the single radial immunodiffusion test.
A qualitative latex agglutination test for equine SAA levels was established by mixing serum with a 1% latex suspension containing 0.2mum particles coated with affinity-purified antibody at 250ng/cm^2 latex. The agglutination was performed on a glass slide at room temperature after a 60-minutes incubation. Standardization of the assay employed a SAA-enriched high density lipoprotein as the primary standard. This test is reliable and reproducible. The results correlate with those of the sandwith ELISA (r=0.953). This test proved to be a simple, rapid and practical assay in clinical laboratories for measuring SAA in field horse serum. Less
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