A preclinical study using recombinant anti-folate receptor-beta immunotoxin for the treatment of rheumatoid arthritis
Grant-in-Aid for Scientific Research (C)
|Allocation Type||Single-year Grants |
|Research Institution||Kagoshima University |
MATSUYAMA Takami Kagoshima University, Graduate School of Medical and Dental Sciences, Professor, 大学院医歯学総合研究科, 教授 (30145479)
NAGAI Taku Kagoshima University, Graduate School of Medical and Dental Sciences, Research Associate, 大学院医歯学総合研究科, 助手 (90363647)
田中 将志 鹿児島大学, 大学院・医歯学総合研究科, 助手 (60381167)
松下 格司 鹿児島大学, 医学部・歯学部附属病院, 助手 (00363630)
|Project Period (FY)
2005 – 2006
Completed (Fiscal Year 2006)
|Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥3,000,000 (Direct Cost: ¥3,000,000)
|Keywords||Immunotoxin / Folate receptor / Rheumatoid arthritis / SCID / Macrophages / Activated fibroblast-like cells / Osteoclasts / SCID|
OBJECTIVE: To investigate the effects of the recombinant immunotoxin dsFv anti-folate receptor beta (FRβ) on the activation and proliferation of the cell that function in inflammatory and degradative processes in rheumatoid arthritis synovial tissue.
1) Productior of recombinant type anti-human FRβ immunotoxin: The Ig VH-PE38 fusion protein and the Ig VL protein were produced in Escherichia coli, and then joined with a disulfide bond by engineering cysteine residues in the framework regions of these proteins.
2) In vitro study: The immunotoxin significantly induced the apoptosis of FRβ-transfected macrophages and adherent RA synovial mononuclear cells.
3) In vivo study: Administration of immunotoxin reduced the numbers of CD68^+ macrophages, activated fibroblast-like cells (VCAM-1^+, Angiopoietin-1^+), inflammatory cytokines (TNF-α, IL-6), angiogenesis (CD34^+ cells) in RA synovial tissue engrafted into SCID mice.
1) Tissue localization of FRβ in rhesus monkey: The liver and skin weakly expressed FRβ in rhesus monkey.
2) Productior of rat-anti mouse FRβ monoclonal antibody: Murine FRβ cDNA was transfected into rat mastcytorra, and injected into foot-pad of WKY rats. Nine monoclonal antibodies were obtained. Any monoclonal antibodies were not cross-reacted with mouse FRα.
3) Tissue localization of FRβ in mouse: FRβ were expressed in thioglycolate-induced peritoneal macrophages (++++), liver (+ 〜 ++), lung (+ 〜 ++) and colon (+), suggesting that inflammatory stimulation may up-regulate; FRβ^+ cells in mouse and rhesus monkey.
Conclusion: Immunotoxin would be a promising tool for the treatment of RA. Prospects: Now, investigators are producing anti-mouse FRβ immunotoxin and will examine the effects on murine arthritis model (CIIA-induced arthritis and serum transfer-induced arthritis).
Report (3 results)
Research Products (4 results)