Dynamics of proinflammatory cytokine expression in the joints of mice with collagen-induced arthritis (CIA)

Abstract

SUMMARYThis report contains a description of the cellular localization and kinetics of proinflammatory cytokine expression in murine CIA, a model for rheumatoid arthritis. Tissue cryostat sections of undecalcified paws from type II collagen-immunized DBA/1 mice, taken 1-10 days after the onset of clinical arthritis, were examined for the presence of tumour necrosis factor-alpha (TNF-a), IL-1b and IL-6 using an indirect immunoperoxidase technique. In parallel, interferon-gamma (IFN-g) production by lymph node cells, stimulated in vitro with type II collagen, was assessed as a marker of T cell activity. The main areas of TNF-a, IL-1b and IL-6 expression were in the synovial lining layer and in tissue contiguous with cartilage and bone (the marginal zone), in particular at sites of pannus formation and joint erosion. There was a progressive increase in the number of TNF-a-, IL-1b-and IL-6-positive cells from day 1 to day 10 of arthritis, during which time IFN-g production by CD4+ T cells from draining lymph nodes declined sharply. A further finding of potential significance was that TNF-a was consistently detected at day 1 of arthritis, whereas IL-1b-positive cells were not found until day 3, suggesting that the expression of TNF-a precedes that of IL-1b.

Indoleamine 2,3 dioxygenase–mediated tryptophan catabolism regulates accumulation of Th1/Th17 cells in the joint in collagen‐induced arthritis

“…Although there is a background level of IDO expression in T cells, dendritic cells are the cells responsible for IDO up-regulation during arthritis, which is consistent with previous findings in other systems (11,26,27). The kinetics of expression follows that of IFN␥ (28) and indicates that IDO is induced by inflammatory stimuli in dendritic cells as one of the feedback mechanisms used by this cytokine to downregulate inflammation. This interpretation is supported by our finding that inhibition of IDO by 1-MT after arthritis onset exacerbates CIA.…”

Adenoviral Transfer of Cyclin-Dependent Kinase Inhibitor Genes Suppresses Collagen-Induced Arthritis in Mice

“…5, A-F). In accord with previous observations (27,28), these cytokines were localized primarily in the nuclear regions, but cytoplasmic staining was also found. In contrast, in the synovial tissues from the AxCAp16-and AxCAp21-treated joints, the staining of IL-1␤ and TNF-␣ was very faint (Fig.…”

“…Saponin permeabilizes the membranes of cells and intracellular organelles, thereby allowing the detection of intracellular cytokines. This technique gives positive nuclear staining in conventional immunohistochemical analyses, thus showing that cytokines can be stained mostly around the nucleus (27,28). The sections were then incubated with 10% normal goat serum for 1 h at room temperature and treated with rabbit anti-human IL-1␤ Ab (LP-712; Genzyme, Cambridge, MA), rabbit anti-mouse TNF-␣ Ab (IP-400; Genzyme), or normal rabbit serum overnight at 4°C.…”

Evaluation of TNF-α and IL-1 Blockade in Collagen-Induced Arthritis and Comparison with Combined Anti-TNF-α/Anti-CD4 Therapy

“…Joints from treated arthritic mice were analyzed for TNF-␣ and IL-1␤ expression using a previously described procedure (30). In brief, joints were embedded in OCT embedding matrix, snap-frozen, and stored at Ϫ70°C until use.…”