Angiostatin gene transfer as an effective treatment strategy in murine collagen‐induced arthritis

Abstract

Objective. To determine the efficacy of local therapy with human angiostatin gene in murine collageninduced arthritis (CIA).Methods. DBA/1 mice were immunized with bovine type II collagen. Before the onset of arthritis, NIH3T3 fibroblasts, transduced with angiostatinexpressing retroviral vectors or control vectors, were transplanted into the knee cavity. The incidence of arthritis in the knee joints was evaluated histologically based on pannus formation and cartilage destruction. Paws were evaluated macroscopically for redness, swelling, and deformities and immunologically for levels of interleukin-1␤. Angiogenesis in paws and knee joints was studied by immunohistochemistry using anti-CD31 antibody and measurement of von Willebrand factor levels.Results. Pannus formation and cartilage erosion were dramatically reduced in knees transplanted with angiostatin-expressing cells. In addition, the onset of CIA in the ipsilateral paws below the knees injected with the angiostatin gene was significantly prevented. Furthermore, angiostatin gene transfer inhibited arthritisassociated angiogenesis.Conclusion. Local production of angiostatin in the knee was able to prevent the onset of CIA not only in the knee injected with genetically engineered cells, but also in the uninjected ipsilateral paw. This suggests that transfer of the angiostatin gene, and potentially also its protein, may provide a new, effective approach to the treatment of rheumatoid arthritis.

Adenoviral delivery of soluble VEGF receptor 1 (sFlt-1) abrogates disease activity in murine collagen-induced arthritis

“…[22][23][24][25][26] Moreover, retrovirally delivered angiostatin also reduced pannus formation and neovascularization. 27 Other studies have reported that endostatin, a 20 kDa fragment of collagen XVIII, can also reduce disease. Intrasynovial injection of human recombinant endostatin reduced both the volume of grafted synovium and the number of vessels in SCID mice engrafted with human RA synovium.…”

Protection against collagen-induced arthritis by intramuscular gene therapy with an expression plasmid for the interleukin-1 receptor antagonist

“…41 On day 21, the animals were boosted with an intradermal injection of 100 mg type II collagen emulsified in Freund's incomplete adjuvant (GIBCO BRL, NY, USA). Gradual onset of arthritis normally starts approximately 4 weeks after initial immunization.…”

Electro-gene therapy of collagen-induced arthritis by using an expression plasmid for the soluble p75 tumor necrosis factor receptor-Fc fusion protein

“…43 On day 21, the animals were boosted with an intradermal injection of 100 mg type II collagen emulsified in Freund's incomplete adjuvant (GIBCO BRL, NY, USA). At 2 days after the boosting, the mice were divided into two groups and individually treated with either 15 mg of plasmid DNA containing cDNA for sTNFR:Fc or control plasmid.…”

“…Levels of vWF were expressed as a percentage, in which the detected vWF was compared with that present in a human plasma calibration standard arbitrarily assigned a value of 100%. 43 The levels of cytokines were normalized to the total amount of protein prepared from tissue lysates, as measured by way of a DC protein assay kit (Bio-Rad Laboratories, Hercules, CA, USA).…”