CIA Mice Research Articles

Therapeutic effects of TGF-β-induced iTregs on established autoimmune diseases (115.3)

Abstract While recent studies reveled injection of nTregs has less therapeutic effects on established autoimmune diseases, it is less clear if iTregs can treat these diseases. We now reveal that unlike nTregs, injection of iTregs can markedly ameliorate established autoimmune arthritis and lupus. Both antigen-specific and polyclonally induced iTregs suppressed the established CIA. CIA mice given iTregs had significantly lower clinic scores than control groups. We found nTregs but not iTregs were converted into Th1/Th17 cells in vitro and in vivo in the inflammatory milieu, injection of iTregs to naïve and immune deficient mice displayed similar levels of Foxp3 stability as comparing with nTregs. Of note, the stability of Foxp3 expression was only found in iTregs during established CIA. iTregs suppressed Th17 and osteoclast differentiation. Injection of iTregs to established lupus decreased levels of anti-dsDNA and proteinuria, and prolonged lupus survival. Anti-TGF-β Ab or TGF-βRI inhibitor, or anti-IL-10R Ab abolished the therapeutic effects of iTregs on lupus. We further observed that DC isolated from lupus received iTregs expressed lower levels of B7.1/7.2 and adoptive transfer of these DCs to another lupus mouse can suppress the disease development. We suggest that iTregs are stable and able to target DC in the inflammatory milieu, these DC then have become tolerogenic DC and further suppress disease progression via its direct or indirect effect in autoimmune disease settings.

A novel series of IKKβ inhibitors part II: Description of a potent and pharmacologically active series of analogs

A novel series of (E)-1-((2-(1-methyl-1H-imidazol-5-yl) quinolin-4-yl) methylene) thiosemicarbazides was discovered as potent inhibitors of IKKβ. In this Letter we document our efforts at further optimization of this series, culminating in 2 with submicromolar potency in a HWB assay and efficacy in a CIA mouse model.

Th17 cells contribute to the exacerbated autoimmune arthritis in CCR2-deficient mice (34.6)

Abstract Th17 cells have been implicated in the pathogenesis of autoimmune diseases including rheumatoid arthritis (RA). Chemokine receptor CCR2 mediates leukocyte trafficking to sites of inflammation and is considered to be an important proinflammatory factor in RA. However, as an unexpected paradox, mice lacking CCR2 (CCR2-/-) develop an accelerated and enhanced arthritis in the collagen induced arthritis model (CIA). To investigate the underlying mechanism(s), we examined the role of Th17 cells in CCR2-/- mice with CIA. We found that the number of Th17 cells was increased significantly in draining lymph nodes of CCR2-/- CIA mice (845±144) compared to WT controls (289±136, p=0.017). In contrast, Th1 cells were not significantly different between groups (485±171 vs. 516±86 for WT, p=0.88). Consistently, CCR2-/- CIA mice had elevated levels of sera IL-17 (84.4±15.5 pg/ml) and IL-17 mRNA in arthritic joints (3.43±0.61 fold) compared to WT mice (45±9.7 pg/ml in the serum and 0.21±0.05 fold in the joint, p=0.04 and 0.009 respectively). Furthermore, Th17-favored cytokines IL-6 and IL-1β were up-regulated in the serum of CCR2-/- animals (IL-6: 366.1±54.5 pg/ml vs. 58.7±13.3 pg/ml for WT, p=0.0004; IL-1β: 189.6±21.9 pg/ml vs. 106.4±21.3 pg/ml for WT, p=0.01). These data suggest that Th17 cells contribute to the pathogenesis of the aggravated phenotype of autoimmune arthritis in CCR2-/- mice. Mice deficient in CCR2 may serve as a model for evaluations of anti-Th17 therapies against RA.

Enterotoxigenic E. coli CFA/I Fimbriae Abate Collagen-Induced Arthritis (CIA) via Regulatory CD39+ T Cells and IL-35 (47.1)

Abstract Colonization factor antigen I (CFA/I) expressed by Salmonella vaccine vectors can prevent experimental autoimmune encephalomyelitis and CIA attributed to suppression of Th1 and Th17 cells by regulatory and immune deviation mechanisms. We hypothesize that soluble CFA/I fimbriae may confer similar protection against CIA as Salmonella-CFA/I. Single dose, mucosal CFA/I application 14 days post-CIA induction abated clinical development of arthritis similar to Salmonella-CFA/I. IFN-γ, IL-6, and IL-17 responses by CD4+ T cells were suppressed in protected mice via the induction of IL-10 and regulatory FoxP3+ CD4+ T cells. Such regulatory T cells were both CD25- and CD25+, each conferring protection upon adoptive transfer to CIA mice. Further evaluation revealed the protective CD25- CD4+ T cells also expressed CD39, ectonucleoside triphosphate diphosphohydrolase, an enzyme responsible for extracellular ATP hyrolysis. Flow cytometry analysis revealed that CFA/I fimbriae nearly doubled the CD39+ CD4+ T cell subset, ~50% of which were CD25-negative. Moreover, mucosal CFA/I delivery enhanced by 2-fold IL-35+ FoxP3+ CD4+ regulatory T cells. CD39+ CD4+ T cells accounted for most (~70%) of TGF-β production by CD4+ T cells. Upon restimulation, these CFA/I -induced CD4+ T cells did not produce IFN-γ or IL-17, but IL-10 levels were enhanced. In conclusion, CFA/I fimbriae alone can augment regulatory T cells for protection against autoimmune diseases. Work supported by P01 AT-04986.

Liver X receptor agonist prevents the evolution of collagen-induced arthritis in mice

Liver X receptors (LXRs) have been characterized as regulators of macrophage inflammatory pathways. Synthetic LXR agonists inhibit the macrophage response to bacterial pathogens and antagonize the induction of a number of pro-inflammatory genes. The aim of this study was to investigate the preventive effects of synthetic LXR agonist, GW3965, treatment on the evolution of arthritis and inflammatory response in a murine CIA model. Intradermal injection of bovine type II CIA in DBA/1 mice. Along with the induction of CIA, mice were treated with oral GW3965 (0.1, 0.3 or 1.0 mg/kg/day) or vehicle from Day 1 to Day 40. Clinical assessment for arthritis scores and histopathological assessment of joint sections were performed. The expression of inflammatory mediators was evaluated by immunohistochemical staining. Serum pro-inflammatory cytokine levels were determined using ELISA. The CIA incidence was 100% on Day 27 and the severity progressed until Day 35 with histological features of cartilage erosion in vehicle-treated mice. GW3965 treatment significantly reduced the arthritis incidence and attenuated the clinical and histological severity, compared with vehicle-treated mice. GW3965 treatment also significantly reduced inflammatory mediator production in joint sections and serum pro-inflammatory cytokine levels in a dose-dependent manner. These results indicate that activation of LXRs suppresses the onset of CIA and reduces inflammation and joint destruction in CIA mice. The data could suggest that LXR treatment is an effective prophylactic approach to suppress the evolution of synovitis and resultant joint destruction observed in RA.

Inhibitory Effect of a Decoction of Eucommiae ulmides OLIVER and Dipsacus asperoides C. Y. Cheng et T.M.Ai on Collagen II-induced Arthritis Mice

Objectives : The object of this study was to verify the inhibitory effect of a decoction of Eucommiae ulmides OLIVER (EU) and Dipsacus asperoides C. Y. Cheng et T.M.Ai (DA) on Collagen II-induced Arthritis Mice (CIA mice). Methods : DBA/1OlaHsd mice were immunized with bovine type II collagen. Boostnig same collagen 21 days later, arthritis was induced and then administrated orally the extract of EU+DA (200 or 50 mg/kg) once a day for 4 weeks and compared with that of methotrexate (MTX, 0.3 mg/kg) as a positive control. Results : Administration of EU+DA suppressed the inflammatory progression of CIA mice and the results were 1. Arthritis index of CIA mice was decreased. 2. EU+DA decreased the production of TNF-, IL-6, IL- in the serum of CIA mice. 3. EU+DA decreased the level of IgM. 4. EU+DAincreasaed , , /CD25 but decreased , (NKT), (NK), in PBMC of CIA mice. 5. EU+DA decreased , , of paw joint in CIA mice. 6. EU+DA decreased subsynovial inflammation. Conclusions : This results demonstrated that extract of EU+DA suppressed the inflammatory progression of CIA mice and supported further studies are required to clarify a mechanism of therapeutic role.

Altered peptide ligands regulate type II collagen-induced arthritis in mice

We reported that peripheral blood mononuclear cells from HLA-DRB1*0101 Japanese patients with rheumatoid arthritis (RA) were highly reactive to 256–271 peptide of type II collagen (CII). Similar to RA, T cells reactive to CII (AA256–271) play a crucial role in the generation of arthritis in CII-induced arthritis mouse (I-Aq). In the present study, we regulated the CII reactivity of T cells from CIA mouse with I-Aq by altered peptide ligand (APL). Eight different APLs were designed and screened for their antagonistic activity using CII reactive cytokine production assay. Four APLs of CII 256–271 exhibited antagonistic activity in CII-reactive T cells. Moreover, intraperitoneally injected APL-5 (G262A) significantly suppressed CII-induced arthritis in mice, whereas the other three APLs did not. Compared with the control, APL-5 suppressed interleukin (IL)-17 production by T cells from CII-induced arthritis mice. These results suggest that CII APL is a potentially suitable therapeutic strategy for the control of RA.

Altered peptide ligands regulate type II collagen-induced arthritis in mice

We reported that peripheral blood mononuclear cells from HLA-DRB1*0101 Japanese patients with rheumatoid arthritis (RA) were highly reactive to 256-271 peptide of type II collagen (CII). Similar to RA, T cells reactive to CII (AA256-271) play a crucial role in the generation of arthritis in CII-induced arthritis mouse (I-A(q)). In the present study, we regulated the CII reactivity of T cells from CIA mouse with I-A(q) by altered peptide ligand (APL). Eight different APLs were designed and screened for their antagonistic activity using CII reactive cytokine production assay. Four APLs of CII 256-271 exhibited antagonistic activity in CII-reactive T cells. Moreover, intraperitoneally injected APL-5 (G262A) significantly suppressed CII-induced arthritis in mice, whereas the other three APLs did not. Compared with the control, APL-5 suppressed interleukin (IL)-17 production by T cells from CII-induced arthritis mice. These results suggest that CII APL is a potentially suitable therapeutic strategy for the control of RA.

Recent development of nanomedicine for the treatment of inflammatory diseases

Recent advance of nanotechnology enables us to develop drug delivery system using nanocarriers. Here I focused on the treatment of inflammatory diseases using nanotechnology, although the most of the pipelines have been developed for cancer treatment so far. First, I described the basic characteristics of nanocarries including liposomes, polymeric micelles, and nanoparticles. Then I showed the therapeutic activity of betamethasone phosphate and FK506 encapsulated in biocompatible and biodegradable blended nanoparticles of PLGA/PLA homopolymers and PEG-block-PLGA/PLA copolymers (stealth nanosteroid or nano-FK) in experimental arthritis models. Various stealth nanosteroids with a size of 45-115 nm were prepared, and then intravenously administered to rats with adjuvant arthritis (AA rats) and/or mice with anti-type II collagen antibody-induced arthritis (CIA mice). The accumulation of stealth nanoparticles with Cy7 in inflamed joints was determined using an in vivo imaging system. The observed strong therapeutic benefit obtained with the stealth nanosteroid/nanoFK in experimental arthritis may have been due to prolonged blood circulation and targeting to the inflamed joint in addition to its sustained release in situ.

Potential analgesic and anti-inflammatory activities of Panax ginseng head butanolic fraction in animals

The present study reports the potential anti-rheumatoid activity of Panax ginseng head part. P. ginseng-head part BuOH fraction (PGHB) was safe in acute toxicity (LD 50 > 5000 mg/kg) and inhibited the partially acetic acid-induced writhes (approximately 32%, P < 0.05) in mice. PGHB (500 mg/kg) inhibited the acetic acid-induced extravasation of Evan’s blue dye in mice by approximately 20.6% ( P < 0.05), and was similar to the suppressive effect of ibuprofen (27.7%) as a positive control drug. Also, PGHB reduced the carrageenan-induced paw edema at 3 h after oral administration, and suppressed the production of serum IL-6 in CIA mice. This suggests that PGHB has potential analgesic and anti-inflammatory activities, and will be the supporting evidence for the potential anti-rheumatoid activity of Korean P. ginseng-head.

The potential analgesic and anti-inflammatory activities of Panax ginseng head BuOH fraction in vivo

Panax ginseng (P. ginseng) has been widely used for many traditional medicinal therapies in Korea and Japan. Oral administration of P. ginseng-head BuOH fraction (PGHB) was found to be safe in acute toxicity (LD50&gt;5,000mg/kg). PGHB inhibited the partially acetic acid-induced writhes (approximately 32%, P&lt;0.05) in mice. This result indicates that PGHB possess antinociceptive activity in chemical-induced pain test models. PGHB (500mg/kg) strikingly inhibited acetic acid-induced extravasation of Evans blue dye in mice by approximately 20.6% (P&lt;0.05), and similar to suppressive effect of ibuprofen (27.7%) as a positive control drug. Also, PGHB reduced approximately 4.55% of carrageenan-induced paw edema at 3h after oral administration. In CIA mice, PGHB suppressed the production of serum IL-6. This suggests that it is concluded that PGHB has potential analgesic and anti-inflammatory activities, and will be supporting the evidence for the potential anti-rheumatoid activity of Korean P. ginseng head.

Sa.11. Amebic Peptide Reduces Inflammation in CIA Mice Model by Down-regulation of Adhesion Molecules and Matrix Metalloproteinases

Sa.11. Amebic Peptide Reduces Inflammation in CIA Mice Model by Down-regulation of Adhesion Molecules and Matrix Metalloproteinases

関節炎モデルにおける血中ピリジノリン測定の有用性

Urinary pyridinoline (U-Py) is a useful marker of bone and cartilage loss in rheumatoid arthritis (RA) . The serum pyridinoline (S-Py) concentration instead of U-Py was measured both in rats with adjuvant-induced (AA) and mice with collagen-induced (CIA) arthritis by high perfomance liquid chromatography (HPLC) . On the seventh day after adjuvant innoculation, the AA rats showed a significant increase in S-Py levels. The level of S-Py was gradually increased with the development of AA. Moreover, in CIA mouse, another arthritic model, on sixty days after innoculation of the type II collagen the S-Py level was also significantly higher than that in the normal mice.These findings indicate that the measurement of S-Py instead of U-Py may be useful as marker of bone and cartilage destruction in patients with rheumatoid arthritis.

Interleukin 1 mediated acceleration of type II collagen-induced arthritis: Effects of anti-inflammatory or anti-arthritic drugs

We previously demonstrated that treatments with rIL-1 beta accelerated the onset and progression of CIA in mice. In the present study, it was observed that IL-1 also enhanced the development of CIA in rats. Like the mouse model, maximal incidence (80-100%) of arthritis occurred within 7 days after the first treatment with IL-1 in rats. Thus, the acceleration of CIA by IL-1 (IL-1 CIA) may be an improved model for the rapid screening of anti-inflammatory and/or anti-arthritic drugs. As a first step to determining the utility of the IL-1 CIA model as a drug screen, we examined the ability of various known anti-inflammatory and anti-arthritic drugs to modify the IL-1 mediated enhancement of CIA in both rats and mice. The results of these studies showed that when analyzed in the IL-1 CIA model, rats and mice exhibited differences in their responses to several of these drugs. For example, dexamethasone, cyclophosphamide, azathioprine, various non-steroidal anti-inflammatory drugs (NSAIDs) as well as methotrexate were found active in the IL-1 CIA of rats. By contrast, the NSAIDs were found to be less effective in suppressing the IL-1 accelerated disease in mice. In both rats and mice, cyclosporine A and several disease modifying anti-arthritic drugs failed to the prevent the development of CIA that was potentiated by IL-1. Thus, in the IL-1 CIA model NSAIDs appeared to be less active in mice than rats. In conclusion, because of the shorter latent period required for the development of arthritis in the IL-1 treated animals, the IL-1 accelerated CIA model in both mice and rats may be useful for screening anti-inflammatory or anti-arthritic compounds.