Characterization of MCP-1 Transcriptional regulation basic investigation of possible gene therapy for arthritis using animal model
| Project/Area Number |
12670433
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
内科学一般
|
|---|
| Research Institution | Yokohama City University |
|---|
Principal Investigator |
ISHIGATSUBO Yoshiaki Yokohama City Univ., Medicine, Professor, 医学部, 教授 (40137039)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
UEDA Atsuhisa Yokohama City Univ., Medicine, Assistant, 医学部, 助手 (60295483)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2000: ¥2,900,000 (Direct Cost: ¥2,900,000)
|
|---|
| Keywords | MCP-1 / transcription / mRNA stability / adenovirus / B-lymphocytes / 単球遊走因子(MCP-1) / 慢性関節リウマチ / 転写制御 |
|---|
| 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.
|
Report
(3 results)
Research Products
(15 results)
