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. 2003 Aug;62(8):707–714. doi: 10.1136/ard.62.8.707

TNF receptor gene therapy results in suppression of IgG2a anticollagen antibody in collagen induced arthritis

P Mukherjee 1, B Wu 1, L Mayton 1, S Kim 1, P Robbins 1, P Wooley 1
PMCID: PMC1754640  PMID: 12860724

Abstract

Background: Therapeutic strategies to block tumour necrosis factor α (TNFα) activity in experimental autoimmune arthritis models and rheumatoid arthritis (RA) have proved highly successful, and provide sustained beneficial effects.

Objective: To examine whether TNFα inhibition has immunological activity beyond the reduction of inflammation in collagen induced arthritis (CIA), an established experimental model of RA.

Methods: Arthritic DBA/1 mice received single periarticular injections of retroviral constructs encoding human TNF receptor (TNF-R) into the affected arthritic paw, at the onset of arthritis. Severity of arthritis, antibodies to collagen type II (CII), and extent of pathological joint damage of arthritic paws were compared between TNF-R and media treated (control) animals 3, 7, 14, 21, and 49 days after disease onset.

Results: Severity of CIA was significantly decreased in TNF-R treated animals compared with controls, 14–34 days after disease onset. Joint destruction was reduced in TNF-R injected joints and in the uninjected contralateral and ipsilateral paws of TNF-R treated animals. Seven days after disease onset, TNF-R treated mice had lower levels of inflammatory Th1 driven IgG2a antibodies to CII (p<0.05) than controls. This altered the anticollagen IgG2a:IgG1 ratio towards Th2 driven IgG1.

Conclusions: Local TNF-R gene therapy in CIA appears to have systemic effects on the anti-CII antibodies. The overall influence of TNF-R gene therapy is that it inhibits the progression of CIA mainly by suppressing the inflammatory Th1 response rather than by stimulating a Th2 response. Therefore, periarticular TNF-R gene therapy may have excellent therapeutic potential in RA.

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Figure 1.

Figure 1

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Effect of a single periarticular delivery of retrovirus mediated MOIN-sTNF-Rc-Ig on the clinical severity of CIA. At arthritis onset 100 µl of media alone (n=24) or 1.6x107 pfu/ml of MOIN-sTNF-Rc-Ig in 100 µl of media (n=22) was injected periarticularly into the affected arthritic paw. Disease activity represented by (A) the mean number of paws affected and (B) the mean paw score was visually scored up to 49 days after disease onset. Data are expressed as mean for each group. SEM <5% of mean (not shown). Statistically significant differences between the two groups is indicated: *p<0.05; #p<0.02). The figure shows representative results from two different individual experiments, which gave similar results.

Figure 2.

Figure 2

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Histopathology of injected, contralateral, and ipsilateral joints. Histological evaluation of control injected (A), contralateral (C), and ipsilateral (E) and MOIN-sTNF-Rc-Ig injected (B), contralateral (D), and ipsilateral (F) arthritic joints were conducted at 49 days after disease onset and at the start of treatment. Media injected control joints show severe synovial inflammation with clear pannus attachment and erosions extending into the subchondral bone, indicating destructive erosive disease. Similar pathological changes are seen in the control arthritic contralateral and ipsilateral joints. Injected, contralateral, and ipsilateral arthritic joints of MOIN-sTNF-Rc-Ig treated mice show a lesser degree of both synovial inflammation and destructive disease, indicating a relatively normal joint architecture. Haematoxylin and eosin staining (magnification x100) used for all sections. JS, joint space; SCB, subchondral bone; CPJ, cartilage/pannus junction; Syn, synovium; P, pannus.

Figure 3.

Figure 3

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Histological scoring of arthritic joints. Arthritic joints were analysed by histology using haematoxylin and eosin staining for extent of joint damage and assigned a histological score (as described in "Materials and methods"). Each column represents the mean scores (SEM) for the total number of mice in each group. *p < 0.05; **p<0.01.

Figure 4.

Figure 4

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Effect of periarticular delivery of MOIN-sTNF-Rc-Ig on the antibody profile to bovine CII. At arthritis onset mice received single periarticular injections of MOIN-sTNF-Rc-Ig or media into the affected arthritic paw. Serum samples were collected 3, 7, 14, 21, and 49 days after arthritis onset. Titres of antibodies to bovine CII were measured by ELISA. Each column represents the anti-bovine CII (A) immunoglobulin; (B) IgG; (C) IgG1; and (D) IgG2a levels in the treated or control sera at these times. Values are means (SEM) for the total number of mice in each group for each time. n=5 for both the media and MOIN-sTNF-Rc-Ig treated mice. *p<0.05.

Figure 5.

Figure 5

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Effect of a single periarticular delivery of MOIN-sTNF-Rc-Ig on the antibovine CII IgG2a:IgG1 ratio. Titres of anti-bovine CII IgG1 and IgG2a were measured by ELISA. The anti-CII IgG2a:IgG1 ratio 3, 7, 14, 21, and 49 days after disease onset is represented. Each column represents the mean ratios (SEM) for the total number of mice in each group for each time. n = 5 for both the media and MOIN-sTNF-Rc-Ig treated mice. **p<0.005.

Figure 6.

Figure 6

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Effect of a single periarticular delivery of MOIN-sTNF-Rc-Ig on the local TNFα levels. Local TNFα levels present in tissue lysates obtained from (A) injected (B) ipsilateral paws of control and MOIN-sTNF-Rc-Ig treated mice, harvested at 3, 7, 14, 21, and 49 days after disease onset were measured by ELISA. Contralateral joints showed results very similar to the ipsilateral joints. The cumulative levels of local TNFα expression in the injected, ipsilateral, and contralateral joints of the treated and control animals are represented in (C). Each column represents the mean (SEM) TNFα levels for the total number of mice in each group for each time. n = 5 for both the media and MOIN-sTNF-Rc-Ig treated mice. *p<0.05.

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