| Project/Area Number |
13670269
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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 |
Bacteriology (including Mycology)
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
UCHIJIMA Masato Dept. of Microbiology, Research Associate, 医学部, 助手 (20252174)
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| Co-Investigator(Kenkyū-buntansha) |
AOSHI Taiki Dept. of Microbiology, Research Associate, 医学部, 助手 (10324344)
NAGATA Toshi Dept. of Microbiology, Associate Professor, 医学部, 助教授 (90275024)
KOIDE Yukio Dept. of Microbiology, Professor, 医学部, 教授 (30126809)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2001: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | CpG-DNA / gene expression / subtraction / immune regulation / 転写調節 |
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| Research Abstract |
Bacterial DNA and immunostimulatory oligodeoxyribonuceotides (ISS-ODN) can induce macrophages, monocytes, and dendritic cells to secrete proinflammatory cytokines such as IL-6, IL-12 and TNF-a, and up-regulate MHC class II, CD40 and CD86 expressions. To further characterize molecular consequences of the recognition of a specific pattern in bacterial DNA, we used a sensitive PCR-based subtraction approach, termed suppression subtractive hybridization (SSH), to isolate genes upregulated in ISS-ODN-stimulated murine spleen cells.
Sequence analysis of 87 clones isolated by SSH revealed that 41 clones were found to correspond exactly to known mouse protein in the BLAST program. Six clones were revealed to have 80-90% homology with known human or rat proteins. Thirty-one clones showed homology with known cDNA whose gene products are not allocated to any functions. Remaining 9 clones showed no significant homology with any sequences in the public database.
Overexpression of NF-kB p105, IFN regulatory factor-1 (IRF-1), proteasome activator (PA) 28b, IRG2 (IFN-IP), and MyD88, which have not previously been reported as overexpressed in the ISS-ODN-stimulated cells, were confirmed by using RT-PCR.
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