Basic studies for development of inhibitor to target nuclear inflammatory cytokine related with sepsis
| Project/Area Number |
18590100
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Drug development chemistry
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| Research Institution | Kagoshima University |
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Principal Investigator |
ITO Yuji Kagoshima University, Faculty of Engineering, Associate Professor (60223195)
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| Co-Investigator(Kenkyū-buntansha) |
SUGIMURA Kazuhisa Kagoshima University, Faculty of Engineering, Professor (80127240)
ABEYAMA Kazuhiro Kagoshima University, Graduate School of Medical and Dental Sciences, Research Assistant Professor (30284897)
HASHIGUCHI Shuhei Kagoshima University, Faculty of Engineering, Assistant Professor (40295275)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,830,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥330,000)
Fiscal Year 2007: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | HMGB-1 / human antibody phage library / sepsis / THP-1 / IL-8 / inflammation / 抗体医薬 / 阻害剤 / ペプチド |
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| Research Abstract |
The purpose of this search is the design of antibody or peptide inhibitor against HMGB-1(High Mobility Group Box Protein-1) which is considered to be a factor causing the sepsis to elucidate the role of the HMGB-1 in inflammation, exacerbation and lethality in sepsis. By using antibody phage display library constipated by us, we isolated two single chain Fv (scFv) antibodies specific to HMGB-1. These antibodies bound to B-domain of HMGB-1 with Kd values of 30 and 47nM, and one of these scFv formed the dimer called diabody. We further estimated the inflammation-blocking activity of the isolated scFv by using IL-8 releasing assay on THP-1 cell which was stimulated with LPS. Unexpectedly, these scFv could not inhibit the IL-8 release but the diabody rather enhanced it. These results indicate that the isolated HMGB-1-specific two scFv don't possesses the inhibitory activity to repress the inflammation which is estimated by IL-8 release from THP-1 and that the diabody has the potential to enhance the IL-8 release probably through the dimerization of HMGB-1, although its mechanism has been still unknown. The isolation of the HMGB-1-specific peptide by the conventional biopanning from the random peptide library was proven to be very difficult, because of the non-specific binding of the phages to HMGB-1.
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Report
(3 results)
Research Products
(24 results)
