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. 2000;2(4):293-302.
doi: 10.1186/ar104. Epub 2000 May 24.

Gene therapy for established murine collagen-induced arthritis by local and systemic adenovirus-mediated delivery of interleukin-4

S H Kim  1 , C H Evans, S Kim, T Oligino, S C Ghivizzani, P D Robbins
Affiliations

Gene therapy for established murine collagen-induced arthritis by local and systemic adenovirus-mediated delivery of interleukin-4

S H Kim et al. Arthritis Res. 2000.

Abstract

To determine whether IL-4 is therapeutic in treating established experimental arthritis, a recombinant adenovirus carrying the gene that encodes murine IL-4 (Ad-mIL-4) was used for periarticular injection into the ankle joints into mice with established collagen-induced arthritis (CIA). Periarticular injection of Ad-mIL-4 resulted in a reduction in the severity of arthritis and joint swelling compared with saline- and adenoviral control groups. Local expression of IL-4 also reduced macroscopic signs of joint inflammation and bone erosion. Moreover, injection of Ad-mIL-4 into the hind ankle joints resulted in a decrease in disease severity in the untreated front paws. Systemic delivery of murine IL-4 by intravenous injection of Ad-mIL-4 resulted in a significant reduction in the severity of early-stage arthritis.

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Figures

Figure 1
Figure 1
Local delivery of Ad-mIL-4. Three days after lipopolysaccharide treatment collagen-immunized mice were injected periarticularly with 5×108 particles of Ad-mIL-4, Ad-eGFP, or saline. (a) Arthritis index. Subsequently, severity of disease was assessed every other day using an established macroscopic scoring system ranging from 0 to 4: 0, normal; 1, detectable arthritis with erythma; 2, significant swelling and redness; 3, severe swelling and redness from joint to digit; and 4, maximal swelling with ankylosis or necrosis. The average macroscopic score was expressed as a cumulative value for all paws, with a maximum possible score of 16 per mouse (n = 20). *P <0.01. (b) Paw swelling. The thickness of each paw was also evaluated using a spring-load caliper. The paw swelling for each mouse was calculated by adding the four thicknesses of the individual paws. *P <0.1;**P <0.05. (c) Number of arthritic paws. At the same time, the total number of arthritic paws were determined for each experimental group (n = 10, maximum 40 per group). *P <0.01.
Figure 2
Figure 2
Histological analysis of the effect of local Ad-mIL-4 treatment in CIA. (a) Ankle joints of mice were isolated from CIA and same-aged normal DBA mice 28 days after adenovirus injection. Ankle joint tissues were stained with hematoxylin and eosin and showed 100× magnification. Particles of Ad-IL-4 or Ad-eGFP (5×108) were injected into ankle joints of mice with established CIA. (b) The joint tissue sections were evaluated in a blinded manner and scored as follows: 1, synovial cell proliferation, synovial hypertrophy with villus formation and/or fibrin deposition; 2, inflammation, synovitis and/or generalized inflammation; 3, cartilage disruption, chondrocyte degeneration and/or ruffling of cartilage surface and/or dystrophic cartilage; and 4, joint destruction, cartilage erosion with abundant inflammation and pannus formation with bone erosion. A total of five joints per group were evaluated by at least two individuals in a blinded manner.
Figure 3
Figure 3
Periarticular injection of Ad-mIL-4 into the hind leg ankle joints conferred a therapeutic effect in the untreated, front paws. The arthritic mice injected in the hind leg ankle joints with Ad-mIL-4, Ad-eGFP or saline were evaluated for the severity of arthritis in either the hind, treated paws (a) or front, untreated paws (b). The macroscopic score (mean ± standard deviation) is expressed as a cumulative value for the two paws, with a maximum possible score of 8.
Figure 4
Figure 4
Induction of IL-10 by Ad-mIL-4 administration. Particles of Ad-IL-4, Ad-IL-10, or Ad-eGFP (5×108) were injected into the ankle joints of naïve DBA mice. The animals were killed at the indicated time points and joint tissue homogenized. The lysate was then monitored for (a) murine IL-4 and (b) murine IL-10 using the appropriate ELISA.
Figure 5
Figure 5
Levels of expression of IL-4 and IL-10 in immunized mice after periarticular injection of diferent doses of Ad-mIL-4. Increasing doses of Ad-mIL-4 were injected periarticularly into mice with established arthritis. The animals were killed at day 35, and the joint tissue homogenized and assayed for levels of (a) IL-4 and (b) IL-10. The doses of Ad-mIL-4 used were as follows: low, 5×106; mid, 5×107; and high, 5×108.
Figure 6
Figure 6
Systemic delivery of Ad-mIL-4. Adenoviruses were injected into the tail vein 24 h after lipopolysaccharide treatment. (a) Arthritis index. Subsequently, severity of disease was assessed every other day using an established macroscopic scoring system ranging from 0 to 4: 0, normal; 1, detectable arthritis with erythma; 2, significant swelling and redness; 3, severe swelling and redness from joint to digit; and 4, maximal swelling with ankylosis or necrosis. +The average macroscopic score was expressed as a cumulative value for all paws, with a maximum possible score of 16 per mouse (n =10). *P <0.001;**P <0.0005. (b) Paw swelling. The thickness of each paw was also evaluated using a spring-load caliper. The paw swelling for each mouse was calculated by adding the four thicknesses of the individual paws. *P <0.05; **P <0.001. (c) Number of arthritis paws. The total number of arthritic paws were determined for each experimental group (maximum 40 per group). *P <0.001.
Figure 7
Figure 7
Serum IL-4 and IL-10 levels following systemic Ad-mIL-4 delivery Ad-mIL-4 was delivered to mice by tail vein injection 30 days after immunization. A week and 30 days after Ad-mIL-4 and Ad-eGFP injection, the mice were bled and serum levels of cytokines measured by ELISA. (a) IL-4 secretion on day 7, (b) IL-10 secretion on day 7, (c) IL-4 secretion on day 30, and (d) IL-10 secretion on day 30.
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References

    1. Evans CH, Robbins PD. Gene therapy for arthritis. J Rheum. 1994;21:779–782. - PubMed
    1. Robbins PD, Evans CH. Prospects for treating autoimmune and inflammatory disease by gene therapy. Gene Ther. 1996;3:187–189. - PubMed
    1. Suzuki H, Sugiyama E, Tunru IS, et al. Suppressive effect of interleukin-4 (IL-4) on IL-6 production by adherent rheumatoid synovial cells. Clin Immunol Immunopathol. 1993;66:67–72. - PubMed
    1. van Roon JA, van Roy JL, Gmelig-Meyling FH, Lafeber FP, Bijlsma JW. Prevention and reversal of cartilage degradation in rheumatoid arthritis by interleukin-10 and interleukin-4. Arthritis Rheum . 1996;39:829–835. - PubMed
    1. Fiorentino DF, Bond MW, Mosmann TR. Two types of mouse T helper cell. IV. Th2 clones secrete a factor that inhibits cytokine production but Th1 clones. J Exp Med. 1989;170:2081–2095. - PMC - PubMed

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