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Arthritis Res. 1999;1(1):81-91. Epub 1999 Oct 26.
Protection against cartilage and bone destruction by systemic interleukin-4 treatment in established murine type II collagen-induced arthritis.

Joosten LA, Lubberts E, Helsen MM, Saxne T, Coenen-de Roo CJ, Heinegard D, van den Berg WB.

Department of Rheumatology, University Hospital Nijmegen, Nijmegen, The Netherlands. l.joosteeuma.azn.nl

INTRODUCTION: Rheumatoid arthritis (RA) is associated with an increased production of a range of cytokines including tumour necrosis factor (TNF)-alpha and interleukin (IL)-1, which display potent proinflammatory actions that are thought to contribute to the pathogenesis of the disease. Although TNF-alpha seems to be the major cytokine in the inflammatory process, IL-1 is the key mediator with regard to cartilage and bone destruction. Apart from direct blockage of IL-1/TNF, regulation can be exerted at the level of modulatory cytokines such as IL-1 and IL-10. IL-4 is a pleiotropic T-cell derived cytokine that can exert either suppressive or stimulatory effects on different cell types, and was originally identified as a B-cell growth factor and regulator of humoral immune pathways. IL-4 is produced by activated CD4+T cells and it promotes the maturation of TH2 cells. IL-4 stimulates proliferation, differentiation and activation of several cell types, including fibroblasts, endothelial cells and epithelial cells. IL-4 is also known to be a potent anti-inflammatory cytokine that acts by inhibiting the synthesis of proinflammatory cytokines such as IL-1, TNF-alpha, IL-6, IL-8 and IL-12 by macrophages and monocytes. Moreover, IL-4 stimulates the synthesis of several cytokine inhibitors such as interleukin-1 receptor antagonist (IL-1Ra), soluble IL-1-receptor type II and TNF receptors IL-4 suppresses metalloproteinase production and stimulates tissue inhibitor of metalloproteinase-1 production in human mononuclear phagocytes and cartilage explants, indicating a protective effect of IL-4 towards extracellular matrix degradation. Furthermore, IL-4 inhibits both osteoclast activity and survival, and thereby blocks bone resorption in vitro. Of great importance is that IL-4 could not be detected in synovial fluid or in tissues. This absence of IL-4 in the joint probably contributes to the disturbance in the Th1/Th2 balance in chronic RA. Collagen-induced arthritis (CIA) is a widely used model of arthritis that displays several features of human RA. Recently it was demonstrated that the onset of CIA is under stringent control of IL-4 and IL-10. Furthermore, it was demonstrated that exposure to IL-4 during the immunization stage reduced onset and severity of CIA. However, after cessation of IL-4 treatment disease expression increased to control values. AIMS: Because it was reported that IL-4 suppresses several proinflammatory cytokines and matrix degrading enzymes and upregulates inhibitors of both cytokines and catabolic enzymes, we investigated the tissue protective effect of systemic IL-4 treatment using established murine CIA as a model. Potential synergy of low dosages of anti-inflammatory glucocorticosteroids and IL-4 was also evaluated. METHODS: DBA-1J/Bom mice were immunized with bovine type II collagen and boosted at day 21. Mice with established CIA were selected at day 28 after immunization and treated for days with IL-4, prednisolone, or combinations of prednisolone and IL-4. Arthritis score was monitored visually. Joint pathology was evaluated by histology, radiology and serum cartilage oligomeric matrix protein (COMP). In addition, serum levels of IL-1Ra and anticollagen antibodies were determined. RESULTS: Treatment of established CIA with IL-4 (1microgram/day) resulted in suppression of disease activity as depicted in Figure 1. Of great interest is that, although 1 microgram/day IL-4 had only a moderate effect on the inflammatory component of the disease activity, it strongly reduced cartilage pathology, as determined by histological examination (Fig. 1). Moreover, serum COMP levels were significantly reduced, confirming decreased cartilage involvement. In addition, both histological and radiological analysis showed that bone destruction was prevented (Fig. 1). Systemic IL-4 administration increased serum IL-1Ra levels and reduced anticollagen type II antibody levels. Treatment with low-dose IL-4 (0.1 microgram/day) was ineffective in suppressing disease score, serum COMP or joint destruction. Synergistic suppression of both arthritis severity and COMP levels was noted when low-dose IL-4 was combined with prednisolone (0.05 mg/kg/day), however, which in itself was not effective. DISCUSSION: In the present study, we demonstrate that systemic IL-4 treatment ameliorates disease progression of established CIA. Although clinical disease progression of established CIA. Although clinical disease progression was only arrested and not reversed, clear protection against cartilage and bone destruction was noted. This is in accord with findings in both human RA and animal models of RA that show that inflammation and tissue destruction sometimes are uncoupled processes. Of great importance is that, although inflammation was still present, strong reduction in serum COMP was found after exposure to IL-4. (ABSTRACT TRUNCATED)

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11056663&dopt=Abstract

Arthritis Res. 1999;1(2):75-84. Epub 1999 Dec 22.
IFN-gamma production in response to in vitro stimulation with collagen type II in rheumatoid arthritis is associated with HLA-DRB1*0401 and HLA-DQ8.

Berg L, Ronnelid J, Sanjeevi CB, Lampa J, Klareskog L.

Karolinska Hospital, Stockholm, Sweden. louise.bermm.ki.se

STATEMENT OF FINDINGS: IFN-gamma was measured in supernatants after in vitro stimulation of peripheral blood mononuclear cells with collagen type II (CII), purified protein derivative or influenza virus. IFN-gamma production in response to CII was similar in rheumatoid arthritis (RA) patients and healthy control individuals. The IFN-gamma response to purified protein derivative and influenza virus was lower in RA patients, reflecting a general T-cell hyporesponsiveness in RA. After recalculating the response to CII taking this hyporesponsiveness into account the CII response was higher in RA patients, and was associated with human leucocyte antigen (HLA)-DRB1*0401 and HLA-DQA1*0301-DQB1*0302 (HLA-DQ8). Rheumatoid arthritis patients with elevated serum levels of immunoglobulin (Ig)G anti-CII antibodies had lower CII-induced IFN-gamma production than patients with low anti-CII levels. The relative increase in CII-reactivity in RA patients as compared with healthy control individuals, and the association of a higher response with RA-associated HLA haplotypes, suggest the existence of a potentially pathogenic cellular reactivity against CII in RA.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11056667&dopt=Abstract [PubMed - in process]

Arthritis Res. 2000;2(4):293-302. Epub 2000 May 24.
Gene therapy for established murine collagen-induced arthritis by local and systemic adenovirus-mediated delivery of interleukin-4.

Kim SH, Evans CH, Kim S, Oligino T, Ghivizzani SC, Robbins PD.

Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.

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.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11056670&dopt=Abstract

Arthritis Res. 2000;2(4):315-26. Epub 2000 Jun 05.
T cells that are naturally tolerant to cartilage-derived type II collagen are involved in the development of collagen-induced arthritis.

Malmstrom V, Backlund J, Jansson L, Kihlberg J, Holmdahl R.

Section for Medical Inflammation Research, Lund University, Lund, Sweden.

The immunodominant T-cell epitope that is involved in collagen-induced arthritis (CIA) is the glycosylated type II collagen (CII) peptide 256-270. In CII transgenic mice, which express the immunodominant CII 256-270 epitope in cartilage, the CII-specific T cells are characterized by a partially tolerant state with low proliferative activity in vitro, but with maintained effector functions, such as IFN-gamma secretion and ability to provide B cell help. These mice were still susceptible to CIA. The response was mainly directed to the glycosylated form of the CII 256-270 peptide, rather than to the nonglycosylated peptide. Tolerance induction was rapid; transferred T cells encountered CII within a few days. CII immunization several weeks after thymectomy of the mice did not change their susceptibility to arthritis or the induction of partial T-cell tolerance, excluding a role for recent thymic emigrants. Thus, partially tolerant CII autoreactive T cells are maintained and are crucial for the development of CIA.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11056672&dopt=Abstract

Zhonghua Jie He He Hu Xi Za Zhi. 2002 Jul;25(7):403-7.
The effect of extracellular matrix remodeling on airflow obstruction in a rat model of chronic obstructive pulmonary disease.

Li H, Cui D, Ma N, Lu L, Gao Y, Cui X, Wang D.

Division of Respiratory Medicine, The 304th Hospital Chinese PLA, Beijing 100037, China.

OBJECTIVE: To study the nature of extracellular matirx (ECM) remodeling and its role in airflow obstruction in a rat model of chronic obstructive pulmonary disease (COPD), and to observe the role of nacetylcystein (NAC), protein kinase C (H(7)) and TGF-beta monocolonal antibody in the regulation of extracellular matrix remodeling in the airway wall. METHODS: Fifty-three Wistar rats were randomly divided into 5 groups: the healthy control group, the COPD model group, the NAC group, the H(7) group and the TGF-beta monocolonal antibody group. Pathologic study of the airway and lung tissue, lung function test and blood gas analysis were performed. Fibroblasts and lymphocytes of the bronchial wall and alveolar macrophages were counted. Areas of the epithelial layer, the smooth muscle layer and the lamina propria were measured by image analyzer. The level of hydroxyproline in bronchial and lung homogenates was determined by biochemistry method. The serum levels of laminin (LN) and hyaluronic acid (HA) were determined by RIA method. RESULTS: The changes in histopathology, lung function and blood gas in the animal model were similar to those in COPD patients. The collagen, mainly type I collagen, in airway walls was significantly increased. The areas of the epithelial layer (21 114 micro m(2)) and the smooth muscle layer (16 061 micro m(2)) were significantly increased in the COPD model as compared to the control group (13 056 micro m(2) and 6 692 micro m(2), respectively) (P < 0.01). In the drug intervention groups these parameters were significantly decreased compared to the control group. The numbers of fibroblasts (13.6 +/- 4.2), lymphocytes (35.6 +/- 6.4) and alveolar macrophages (14.8 +/- 1.1) in the model group, were significantly increased compared to the control group (6.8 +/- 1.4, 6.1 +/- 1.2 and 3.5 +/- 1.2, respectively) (P < 0.01, 0.001, 0.001), while in the drug intervention groups the cells were significantly decreased except for fibroblasts in the H(7) group. The hydroxyproline level of the model group (111.5 +/- 2.3) pg/ml was significantly increased as compared to the control group (47.8 +/- 9.7) pg/ml (P < 0.05) and was negatively correlated with FEV(0.3)/FVC (P < 0.001) and positively correlated with airflow resistance (P < 0.01). The number of fibroblasts was also positively correlated with the level of hydroxyproline (P < 0.001). The serum levels of LN (26 +/- 4) micro m/L and HA (19.4 +/- 1.4) micro g/L in the model group were significantly increased compared to the control group (15 +/- 3) micro g/L, and (10.9 +/- 2.9) micro g/L, respectively (P < 0.05). Hydroxyproline in the NAC group (83.1 +/- 41.7) pg/ml and the TGF-beta monoclonal antibody group (71.2 +/- 20.3) pg/ml was significantly decreased, while in the H(7) group (160.6 +/- 41.7) pg/ml it was significantly increased. CONCLUSION: Excessive deposition of ECM, mainly of type I collagen, and proliferation of functionally activated fibroblasts were important pathological changes in airway remodeling and the important causes of airflow obstruction. TGF-beta monoclonal antibody and NAC can modulate airway extracellular matrix remodeling. H(7) can increase collagen deposition in the airway wall but the underlining mechanisms need to be elucidated.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12435298&dopt=Abstract [PubMed - in process]

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