| Project/Area Number |
16591002
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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 |
膠原病・アレルギー・感染症内科学
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| Research Institution | University of Occupational and Environmental Health |
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Principal Investigator |
SAITO Kazuyoshi University of Occupational and Environmental Health, School of Medicine, associate professor, 医学部, 助教授 (30279327)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Yoshiya University of Occupational and Environmental Health, School of Medicine, professor, 医学部, 教授 (30248562)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2005: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | regulatory T lymphocyte / autoimmune / FOXP-3 / CD4 / CD25 / FOXP-3 |
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| Research Abstract |
Recent studies have emphasized the importance of T cells with regulatory/suppressor properties in controlling autoimmune diseases. CD4(+) CD25(+) regulatory T(T(reg)) cells play a critical role in the maintenance of peripheral tolerance and the prevention of autoimmunity. We developed anti-FOXP3 polyclonal and employed western blotting analysis and flowcytometric analysis using mononuclear cells from autoimmune diseases.
1.We have explored the characteristics of CD4(+) CD25(+) T(reg) cells or FOXP3 positive cells in patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The frequency and phenotype of CD4(+) CD25(+) T cells as well as FOXP3 positive cells in peripheral blood (PB) from patients with RA, SLE and PBMC from normal controls were analyzed. An decreased frequency of CD4+ T cells expressing CD25 or FOP3 were detected in PB from patients with RA and SLE compared with that in healthy donor.
2.We transfected FOXP3 expression vectors to COST cells by lipofection and peripheral lymphocytes using nucleofector. Then expression of FOXP3 protein in both cells were confirmed by western blotting as well as by FACScan analysis.
3.We also investigate the effect of cytokines or viral protein including human T lymphocytic leukemia virus tax p40 and cytomegalovirus early antigen-on expression of FOXP3 gene. We co-transfecto FOXP3 expression vector and luciferase reporter vector, which was ligated with 3 times tandem repeats of FOXP3 binding sequences to peripheral lymphocytes using nucleofector. Co-transfection of FOXP3 did not induce luciferase activities even in the presence of cytokines including IL-1,2,4,6 and TNF. Co-transfection of Tax p40 or CMV E1+2 expression vector did not increase of luciferase activities.
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