2001 Fiscal Year Final Research Report Summary
Characterization of MCP-1 Transcriptional regulation basic investigation of possible gene therapy for arthritis using animal model
| Project/Area Number |
12670433
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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 |
内科学一般
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| Research Institution | Yokohama City University |
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Principal Investigator |
ISHIGATSUBO Yoshiaki Yokohama City Univ., Medicine, Professor, 医学部, 教授 (40137039)
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| Co-Investigator(Kenkyū-buntansha) |
UEDA Atsuhisa Yokohama City Univ., Medicine, Assistant, 医学部, 助手 (60295483)
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| Project Period (FY) |
2000 – 2001
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| Keywords | MCP-1 / transcription / mRNA stability / adenovirus / B-lymphocytes |
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| Research Abstract |
We previously established that the nuclear factors Sp1 and NF-kB are critically involved in regulating human MCP-1 gene transcription. NF-kB is constitutively activated in B lymphocytes, raising the possibility that MCP-1 is also constitutively expressed by these cells. PCR analysis of resting peripheral blood B cells, however, detected no evidence of MCP-1 mRNA production. In contrast, human B cells incubated for 5 days with a combination of IL-10 plus anti-CD40, responded to LPS and TPA stimulation by significantly increasing MCP-1 mRNA levels. Under the same conditions, NF-kB was not up-regulated, suggesting that the increase in MCP-1 mRNA did not reflect increased rates of gene transcription. Additional studies to identify the mechanism underlying the accumulation of MCP-1 mRNA were performed using the human RPM1 8226 B cell line, which mirrored the response of peripheral blood B cells. LPS and TPA strongly up-regulated MCP-1 mRNA expression in RPM1 8226 cells. Of interest, this treatment did not alter the binding of NF-kB to the MCP-1 5'-flanking sequence, but did prolong the half-life of MCP-1 mRNA by up to 15-fold. These findings indicate that changes in MCP-1 mRNA expression are dependent on post-transcriptional regulation. Current efforts are directed towards investigating the control of MCP-1 expression in vivo. Towards that end, we constructed a recombinant adenovirus encoding expression cassete containing murine MCP-1. This adenovirus has been shown to efficiently infect target cells, which up-regulate expression of the exogenous MCP-1 present in the recombinant virus. Future studies will solve a role of MCP-1 on the pathogenesis of Rheumatoid arthritis.
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Research Products
(2 results)
