Antigen-induced Arthritis In Mice Research Articles

In VivoAssessment of Synovial Microcirculation and Leukocyte–Endothelial Cell Interaction in Mouse Antigen-Induced Arthritis

The microcirculation and leukocyte-endothelial cell interaction in synovial tissue of an inflamed joint are known to play a crucial role in the pathogenesis of rheumatoid arthritis. The aim of this study was to characterize the in vivo changes in the microvasculature and in leukocyte-endothelial cell interactions in the mouse synovial tissue using intravital fluorescence microscopy in three stages of antigen-induced arthritis. The expression of E- and P-selectin and ICAM-1 were also studied using immunohistochemistry.Antigen-induced arthritis (AiA) was produced in Balb/c mice. The severity of arthritis at three different phases was quantified using a clinical and histological score. For the intravital fluorescence microscopy measurements, the patella tendon was partially resected for visualization of the intraarticular synovial tissue of the knee joint. The number of rolling and adherent leukocytes, functional capillary density (FCD) and RBC velocity were quantitatively measured in synovial microvessels. Expression of ICAM-1, E- and P-selectin was assessed by immunohistochemistry.There was a significant increase in the leukocyte rolling fraction in postcapillary venules in the acute phase of AiA (from 0.26 +/- 0.05 in controls to 0.45 +/- 0.04 8 d after AiA induction). The number of leukocytes adherent to the endothelium was significantly elevated in all phases of arthritis (from 121 +/- 27 in controls to 376 +/- 62 mm2 63 d after AiA-induction). Functional capillary density was significantly enhanced in the acute (332 +/- 15 cm/cm2) and intermediate phases (320 +/- 15 cm/cm2) compared to control values (227 +/- 15 cm/cm2). Arthritis resulted in a distinct increase in the expression of ICAM-1 on the synovial endothelium in all phases of AiA. E- and P-selectin expression were detected only in the acute phase.Our model provides new insights into the microcirculatory changes which occur in the synovial tissue of an arthritic joint.

Effect of a Monoclonal Antibody against Interleukin-4 on Suppression of Antigen-Induced Arthritis in Mice by Oral Administration of the Inducing Antigen

We investigated a role for interleukin-4 (IL-4) in suppression of T-cell-mediated antigen-induced arthritis (AIA) in mice by oral administration of the inducing antigen. For this investigation, a monoclonal antibody (11B11 mAb) that specifically neutralizes IL-4 was employed. AIA was induced by immunization with methylated bovine serum albumin (mBSA) (day 0) followed by intraarticular injection of mBSA into the ankle joint. To induce oral tolerance, mBSA was administered orally once a day from days −5 to −1. The 11B11 mAb was injected ip 30 min before each oral administration of mBSA. Oral administration of mBSA resulted in marked suppression of AIA. Suppression of the joint inflammation by the oral antigen was significantly diminished by treatment with 11B11 mAb. The mAb treatment was also followed by blockade of suppression of proliferative responses of lymphoid cells to mBSA by the oral antigen. Furthermore, secretion of IL-4 was significantly increased following oral administration of mBSA and the increased IL-4 secretion was markedly reduced by treatment with 11B11 mAb. There was a decrease in production of IFN-γ in orally tolerized mice that was blocked by the IL-4-neutralizing mAb. Thus, treatment with an anti-IL-4 mAb appears to be effective in blocking suppression of AIA by oral administration of the inducing antigen. The results also suggest that IL-4 may play a role in downregulation of T-cell-mediated inflammation by feeding pathogenic antigens.

Degradation of articular cartilage during the progression of antigen-induced arthritis in mice: A scanning and transmission electron microscopic study

Ultrastructural changes of knee articular cartilage in C57B1/6 mice were studied in the course of antigen-induced arthritis by means of scanning and transmission electron microscopy. First damages of the cartilage surface were seen one hour after arthritis induction. The earliest signs were the loss of the superficial electron-dense layer as well as a progressive loss of proteoglycans in the cartilaginous matrix on the surface. Breaks of collagen fibres were already detected at the first day of arthritis. The chondrocytes of the superficial cartilage layer showed an increase of the intracellular membraneous system in the early phase of arthritis. Thereafter chondrocytes on the surface became more and more necrotic. On the 7th day of arthritis acute alterations of cartilage had developed completely. Many lacunae of chondrocytes were opened and the cartilage surface showed deep structural defects with adhering cells, probably lymphocytes and macrophages. In the following time these destructive processes were demonstrated along with cellular proliferation as a sign of repair attempts in hyaline cartilage.

1306 Axonal regeneration in the carp spinal cord

To identify how the gp130-signaling cytokine oncostatin M (OSM), acting alone or in concert with IL-1β or TNFα, affects synovial fibroblast expression of genes relevant to inflammation and bone erosion in inflammatory arthritis.Synovial fibroblasts (SFs) were isolated from non-arthritic wild type (WT) or OSM receptor deficient (OSMR−/−) mice and stimulated with OSM, IL-1β or TNFα and their combinations. Cytokine gene expression was assessed by quantitative RT–PCR. ELISA, flow cytometry and immunohistochemistry identified protein expression. Gene expression patterns were confirmed in SFs isolated from patients with osteoarthritis (OASFs) and rheumatoid arthritis (RASFs).Expression of OSM and its receptors, gp130, OSMR and LIFR, was increased in synovial tissue from the mouse antigen-induced arthritis model. In isolated WT mouse synovial fibroblasts OSM alone, or in synergy with IL-1β, or together with TNFα, potently induced expression of the pro-inflammatory cytokine IL-6. OSM also induced a sustained increase in mRNA levels of the pro-osteoclastic cytokine RANKL. Combining OSM with IL-1β, but not with TNFα, further increased RANKL expression. Importantly these effects of OSM were all dependent on the expression of OSMR. Furthermore, OSM also increased expression of its own receptors, gp130 and OSMR and the IL-1 receptor, IL1-R1; the latter effects were also observed in both human OASFs and RASFs.Together our data suggests that OSM signaling via OSMR in SFs has the potential to contribute significantly to joint destruction in inflammatory arthritis. It not only induces expression of pro-inflammatory and pro-osteoclastic cytokines but can also augment its own actions and that of IL-1 by inducing expression of OSMR and IL-1R1.

Role of interleukin-1, tumor necrosis factor alpha, and interleukin-6 in cartilage proteoglycan metabolism and destruction. Effect of in situ blocking in murine antigen- and zymosan-induced arthritis.

To determine the involvement of interleukin-1 (IL-1), tumor necrosis factor (TNF), and IL-6 in the cartilage pathology of murine antigen-induced arthritis (AIA) and zymosan-induced arthritis (ZIA). Arthritis was induced by intraarticular injection of zymosan in naive mice or by subcutaneous injection of methylated bovine serum albumin in sensitized animals. Mini-osmotic pumps releasing human recombinant IL-1 receptor antagonist (IL-1ra) protein were implanted intraperitoneally 2 days before arthritis induction, and neutralizing antibodies directed against murine IL-1 alpha, IL-1 beta, TNF alpha, or IL-6 were administered 1 day before. Proteoglycan (PG) synthesis and degradation were assessed in patellar cartilage. Murine IL-1 alpha and IL-1 beta injected intraarticularly at doses of 0.1-100 ng suppressed chondrocyte PG synthesis. The highest dose of TNF tested (100 ng) decreased PG synthesis marginally. In contrast, the maximum dose of IL-6 (1 microgram) stimulated PG synthesis 2 days after injection. Treatment of AIA with neutralizing monoclonal antibodies against either TNF alpha or IL-6 did not reduce either the PG degradation or the suppression of its synthesis. However, treatment with anti-IL-1 (alpha + beta) polyclonal antibodies totally prevented PG suppression, although the initial breakdown of PG was unaffected. This effect was confirmed when IL-1ra was administered in high doses. Moreover, treatment of ZIA with anti-IL-1 (alpha + beta), but not with anti-TNF, resulted in normal PG synthesis, confirming the key role played by IL-1 in the inhibition of PG synthesis. Treatment of AIA with anti-IL-1 did not affect inflammation during the acute phase, but a significant reduction of ongoing inflammation was noted at day 7, and there was a marked reduction in the loss of cartilage PG. The suppression of PG synthesis in both ZIA and AIA in mice is due to the combined local action of IL-1 (alpha + beta), and neither IL-6 nor TNF is involved. Moreover, the normalization of PG synthesis brought about by blocking of IL-1 ameliorates the cartilage damage associated with AIA.

Osteoporosis in rheumatoid arthritis: a molecular biological aspect of connective tissue gene activation.

Osteoporosis, especially the juxtaarticular osteoporosis of involved joints, is a characteristic manifestation of rheumatoid arthritis (RA). Histomorphometric studies suggest the existence of increased bone turnover in RA: impaired bone formation and hightened osteoclasic bone resorption. Recent studies show that important mediators in the pathogenesis of RA such as prostaglandin E, interleukin 1 (IL1) or tumor necrosis factor (TNF) alpha also play important roles in bone remodelling. Prostaglandin E2 promotes maturation of osteoclasts from hematopoietic precursor cells. IL1 inhibits collagen synthesis in osteoblasts. IL1 enhances collagenase and stromelysin gene expression and stimulates osteoclastic bone resorption. TNF alpha inhibits bone collagen synthesis and causes osteoclastic bone resorption. TNF alpha, and possibly IL1, enhances collagenase and stromelysin gene expression by stimulating the AP-1 promoter sites of the genes. Constitutive expression of c-fos induces joint destruction without lymphocyte infiltration in antigen-induced arthritis in mice, and supports cell growth of human rheumatoid synovial cells, possibly acting on the AP-1 sites. Furthermore, constitutive c-fos expression decreases collagen synthesis in osteoblasts and increases the mediator secretion from osteoblasts thereby stimulating osteoclastic bone resorption. These findings suggest that signal transduction through AP-1 transcriptional regulation sites may play an important role in the pathogenesis of joint destruction and osteoporosis in RA.

The effect of an interleukin-1 receptor antagonist protein on type II collagen-induced arthritis and antigen-induced arthritis in mice.

To investigate the anti-arthritic effect of recombinant human interleukin-1 receptor antagonist protein (IRAP) in two experimental models of arthritis. Recombinant IRAP was administered daily to mice with type II collagen-induced arthritis (CIA) or with antigen-induced arthritis (AIA) provoked by methylated bovine serum albumin (mBSA). Disease incidence and severity were assessed by a clinical index and histologic features. Serum antibody to type II collagen, spleen cell proliferation to mBSA, and anti-IRAP antibodies were measured as indices of immune function. IRAP reduced the incidence and delayed the onset of CIA and suppressed the antibody response to type II collagen. In contrast, IRAP did not affect the pathogenesis of AIA and had no effect on either humoral or cellular immune responses to mBSA in arthritic mice. These observations suggest that interleukin-1 may play a prominent role in the development of some, but not all, forms of arthritis.

Humoral and cell-mediated sensitivity to cartilage constituents in mice with antigen-induced arthritis.

Serum antibodies against native collagen type II and cartilage proteoglycans as well as proliferative responses of spleen lymphocytes to these antigens were evidenced during the progression of antigen-induced arthritis in mice. Because there were some correlations to the severity of arthritis, it is suggested that autoimmunity to cartilage constituents play a role for the persistence of the joint inflammation and cartilage destruction in this experimental model of chronic arthritis.

Age-related changes in joint damage caused by antigen-induced arthritis in mice.

Age-related changes in joint damage caused by antigen-induced arthritis in mice.

The role of MHC class II antigens in the flare up reaction of antigen-induced arthritis in mice.

The role of MHC class II antigens in the flare up reaction of antigen-induced arthritis in mice.

Flare of antigen-induced arthritis in mice after intravenous challenge. Kinetics of antigen in the circulation and localization of antigen in the arthritic and noninflamed joint.

Intravenous injection of methylated bovine serum albumin (mBSA) in mice with unilateral, chronic, mBSA-induced arthritis has been shown to cause a flare of smouldering arthritis without affecting the contralateral, noninflamed knee joint. We studied the kinetics of 125I-labeled mBSA in the blood, and the accumulation of antigen in both arthritic and noninflamed joints. The bulk of the antigen was eliminated from the blood within 10-30 minutes, and accumulation of antigen in the joints occurred mainly within this period. The amounts of antigen found in the arthritic joints were of the same order of magnitude as the amounts found in noninflamed joints. Autoradiography of whole joint sections revealed that, in arthritic joints, antigen was located primarily at the deep capillaries and large vessels, and in the noninflamed joints, antigen was located at the small superficial capillaries. Antigen was handled by granulocytes in the arthritic joint and by synovial lining cells in the noninflamed joint. Our data indicate that tiny amounts of antigen reach the synovial stroma in both normal and arthritic joints but cause inflammation only in the arthritic joints, because of local hyperreactivity.

Studies on Antigen-Induced Arthritis in Mice

Abstract Antigen-induced arthritis was developed in mice as a model of human rheumatoid arthritis by using methylated bovine serum albumin (mBSA) as antigen. It was found that most strains were susceptible, whereas CBA mice were resistant. We therefore investigated the humoral and cell-mediated immune responses to mBSA in resistant mice (CBA) and susceptible mice (exemplified by C57BL) to determine whether these were associated with susceptibility to arthritis. The resistant strain (CBA) differed from the susceptible strains in the following respects. First, there was a lower humoral immune response to mBSA as measured by passive hemagglutination, but this could be overcome by a larger immunogenic dose. Secondly, there were differences in response to low doses of DNP-mBSA after mBSA carrier preimmunization. Thirdly, there were striking differences in delayed-type hypersensitivity (DTH) to mBSA as determined by a radioisotopic assay in vivo; the response of CBA mice occurred early, at 5 days, declined quickly, and was weaker, whereas that of C57BL mice developed later and was long sustained. Genetic studies of the DTH response with hybrids and backcrosses showed an oligogenic control of immune responsiveness, with one gene being linked to the H-2b allele of the susceptible C57BL mice, and another being independent of the H-2 complex. Our findings indicate that in mice, susceptibility to antigen-induced arthritis with mBSA correlates with a higher responder state to this antigen, and that T cells are the major if not the only determinant of the high responder state.

Studies on Antigen-Induced Arthritis in Mice

Abstract Antigen-induced arthritis in mice occurs after immunization and subsequent intraarticular challenge with methylated bovine serum albumin (mBSA). In adoptive transfer experiments, susceptible C57BL mice and resistant CBA mice were compared in their capacity to express delayed-type hypersensitivity (DTH) by ear assay, and to express arthritis. The expression of DTH could be transferred incrementally by lymphoid cells in C57BL mice, but not in CBA mice. Both immune lymphoid cells and, to a much lesser extent, serum transferred the capacity to develop arthritis in C57BL mice. The reactivity of transferred cells was abolished by anti-Thy-1 but enhanced by enrichment for T cells with anti-immunoglobulin columns. If this model disease can be equated with human rheumatoid synovitis, the lesions in the human disease would be an expression of a T cell-dependent activity

Antigen-induced arthritis in mice. I. Induction of arthritis in various strains of mice.

Antigen-induced arthritis was established in the mouse by immunizaiton with methylated bovine serum albumin (mBSA) in complete Freund's adjuvant with B pertusis vaccine. The knee joint was injected after 21 days with mBSA in saline. The arthritis was chronic, antigen-specific, and T-cell dependent in hypothmic nu/nu mice. C57BL and balb/c mice were susceptible, whereas CBA mice were relatively resistant. Susceptibility was dominant; one gene was loosely linked to the "b" allele of the H-2 complex of C57BL mice.