Serum IL-4, IL-10 and IL-6 levels in inflammatory arthritis

As the available in vitro and in vivo data suggest that interleukin (IL)-4 and IL-10 have immunosuppressive activity, our hypothesis was that serum IL-4 and IL-10 levels would correlate inversely with parameters of inflammation in patients with inflammatory arthritis. IL-4 was detected in the serum of 12 out of 140 patients with rheumatoid arthritis (RA), which was increased compared to the proportion found with patients with osteoarthritis (OA; P< 0.02). In addition, IL-4 was detected in the serum of 2 of 19 patients with systemic lupus erythematosus (SLE), 2 of 24 patients with psoriatic arthritis and 1 of 5 patients with Behçet's syndrome. No IL-4 was detected in patients with the following conditions: OA (58 patients), gout (17 patients), ankylosing spondylitis (6 patients), Reiter's syndrome (6 patients), polymyalgia rheumatica (6 patients), temporal arteritis (5 patients) and scleroderma (3 patients). No IL-10 was detected in any of the sera tested. We discuss the possible relevance of these results to the regulation of the immune response evident in inflammatory arthritis.     

Ameliorative effect of ozone on cytokine production in mice injected with human rheumatoid arthritis synovial fibroblast cells

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by invasion of hyperplastic synovial cells and progressive joint destruction. Ozone therapy has been proposed as an immunomodulator and cellular metabolic activator which shows long-term anti-inflammatory effects and serves to reduce further the proinflammatory factors. We purified RA synovial fibroblast cells (RA-SFc) from patients and avoided contaminating macrophages by flow cytometry, then treated them with ozone. Following the observable decreased production of proinflammatory factors TNF-, IL-1, and IL-6 from RA-SFc, we infused the cultured RA-SFc into joints of severe combined immunodeficiency mice. The mRNA and protein levels of the RA-SFc exposed to 3% and 5% ozone were the same. As a result, 3% and 5% ozone applied externally ameliorated the inflammatory reaction of RA without toxicity or serious side effects. Therefore, ozone injected into the knees of RA patients could become a valuable treatment, and we confirm the interactive mechanism between ozone and RA-SFc.     

Participation of substance P distribution in the cytokine production of rheumatoid synovium

Based on findings which suggested the involvement of the neuropeptide substance P in the pathogenesis of rheumatoid arthritis (RA), we investigated the mechanism of synovial pannus formation in RA, and examined the interaction between the cytokine production of synovial tissues and the concentration of substance P in the cartilage–pannus junction (CPJ). The CPJ and other peripheral synovial tissues were separately obtained from each part of the synovium from the knee joints of seven RA patients. The concentrations of substance P and the cytokines interleukin (IL)-1β and IL-6 in the CPJ and peripheral synovial tissues were determined by enzyme-linked immunosorbent assays. In addition, synovial cells were isolated from the CPJ and peripheral synovial tissues and treated with substance P or neurokinin-1 receptor antagonist to analyze the changes in cytokine production. The substance P levels were 211.2 and 50.5 pg/mg protein in the CPJ and the peripheral synovium, respectively. The IL-1β and IL-6 levels in the CPJ were 24.6 and 12.8 pg/mg protein, respectively. In the peripheral synovium, these levels were 4.3 and 2.5 pg/mg protein, respectively. In the CPJ, the IL-1β and IL-6 levels in tissue containing a high concentration of substance P (>200 pg/mg protein) were 39.4 and 21.6 pg/mg protein, respectively, and those in tissue containing a low concentration of substance P (≤200 pg/mg protein) were 11.6 and 5.1 pg/mg protein, respectively. Synovial cells from the CPJ produced higher levels of IL-1β and IL-6 than those from peripheral tissues. In addition, treatment of the cells with an NK-1 antagonist significantly reduced the production of these cytokines by the synovial cells. The theory that substance P plays a role in the pathogenesis of RA via the upregulation of cytokine production should be considered in further studies on the immunomodulatory properties of substance P in arthritis. Received: June 17, 1998 / Accepted: February 4, 2000     

DOWN-REGULATION OF FIBRINOGEN BIOSYNTHESIS BY IL-4, IL-10 AND IL-13

High levels of fibrinogen are recognized as an important vascular risk factor; however, it is not known if the increase of plasma fibrinogen is directly responsible for this risk, or is only a marker of vascular inflammation. To support this second hypothesis, Oncostatin M (OSM) is a potent stimulator of fibrinogen biosynthesis and induces smooth muscle cell proliferation. In the same way, we analysed whether interleukin-4 (IL-4), interleukin-10 (IL-10) or interleukin-13 (IL-13), which protect vessel walls from monocytes injuries leading to atherosclerosis, could influence fibrinogen biosynthesis. The two levels of regulation of fibrinogen biosynthesis were tested: firstly, the direct effect of these cytokines on fibrinogen production by the hepatoma cell line Hep G2, and secondly their effect on the secretion of hepatocyte stimulating factor (HSF) activity in the supernatant of lipopolysaccharide (LPS)-activated monocytes. IL-4 and IL-13 added to Hep G2 cells down-regulated both the increase of fibrinogen secretion induced by IL-6 and fibrinogen mRNA levels, this effect being more pronounced when Hep G2 were preincubated with the two cytokines before IL-6 addition. The effect of IL-10 was evidenced only on mRNA expression. IL-10 and IL-13 dose-dependently decrease HSF activity secreted by LPS-activated monocytes, whereas IL-4 had no effect. However, the three cytokines decreased HSF activity when monocytes were incubated with the cytokines before LPS activation. The effects of these cytokines on HSF activity are related to variations of IL-6 and OSM secretion. Our data strengthen the hypothesis that the fibrinogen level is a marker of vascular disease, since cytokines which have a protective vascular effect down-regulate fibrinogen production.     

Tumor necrosis factor-alpha and Interleukin-10 gene promoter polymorphisms in Turkish rheumatoid arthritis patients

Tumor necrosis factor and interleukin 10 have been implicated in the pathogenesis of rheumatoid arthritis (RA). Certain single-nucleotide polymorphisms (SNPs) within the promoter region of the IL-10 and TNF genes have been associated with altered levels of circulating IL10 and TNF. We aimed to explore the association of IL-10 and TNF-alpha polymorphisms in Turkish RA patients. We analyzed the association of TNF-alpha (-308G/A, -238G/A, -376G/A) and IL10 (-1082G/A, -819C/T, -592C/A) polymorphisms in 98 Turkish patients with rheumatoid arthritis and 122 healthy subjects using ARMS-PCR. The correlation of these findings with RF positivity and erosive disease in RA patients was also sought. A significant association was found between having RA and -1082 G allele (p = 0.008; OR = 1.44, 95% CI 1.11-1.86). There was no association between RA and -819C/T polymorphism. Significant differences were observed in IL10 GCC and ACC haplotypes distribution between RA and control subjects (p = 0.006; OR = 1.46, 95% CI 1.13-1.89 and p = 0.011; OR = 1.43, 95% CI 1.09-1.88, respectively). No statistically significant association was found between TNF-alpha 308G/A, -238G/A, -376G/A polymorphisms and RA. No significant association was found between RF positivity and erosive disease and TNF-alpha, IL10 gene polymorphisms. In addition, when combined genotypes were analyzed, no significant difference was found between RA patients and healthy controls. Our findings suggest that IL-10 1082 G/A polymorphism or GCC, ACC haplotypes may be associated with RA in Turkish patients.     

Immunomodulatory effect of Hypoderma lineatum antigens: in vitro effect on bovine lymphocyte proliferation and cytokine production

This study examines the immunomodulatory effect of a crude larval extract (CLE), obtained from first stage larvae (L1) of H. lineatum , and the purified fractions hypodermin A (HyA), HyB and HyC. Proliferative responses of peripheral blood mononuclear cells (PBMC) from uninfested and previously infested cattle and the production of the cytokines IL-10, IL-4 and IFN-γ, in response to concanavalin A (Con A), were determined. The stimulation index of peripheral blood mononuclear cells from uninfested cattle was significantly lower than that from infested animals with the different antigens assayed. The HyA was the antigen that most inhibited the proliferative response, followed by the HyB, the HyC and the CLE. This hypodermin provoked an increase of IFN-γ and a suppression of IL-10 production that would support a Th1-like cytokine response. The HyB reduced the production of IL-10 stimulated by the Con A in cultures from infested animals. The HyC did not modulate the production of cytokines. Finally, the CLE induced a marked suppression in the production of the different cytokines in cultures from naïve and previously sensitized animals. Our results indicate that Hypoderma larval secretions are comprised of different components (hypodermins) that individually induce distinct but partially overlapping modulatory responses.     

Serum levels of IL-10, IL-6, IL-1ra, and sIL-2R in patients with psoriatic arthritis

Our objective was to evaluate the levels of interleukin-6 (IL-6), soluble receptors of IL-2 (sIL-2R), IL-10, and IL-1 receptor antagonists (IL-1ra) in the serum of patients with psoriatic arthritis (PsA) and to assess the correlation between these levels and parameters of clinical activity of skin and joint disease. In total, 34 patients with PsA and ten healthy volunteers participated in the study. Assessment of joint disease included duration of morning stiffness, number of tender and swollen joints, right and left grip, the presence of inflammatory spinal back pain, and Schober test. Current severity of skin disease was graded according to the psoriasis area and severity index (PASI). Erythrocyte sedimentation rate (ESR) was determined as a marker of disease activity. Serum levels of IL-6, sIL-2R, IL-1ra, and IL-10 were measured by an enzyme immunoassay kit. Significantly higher serum levels of IL-6, sIL-2R, IL-1ra, and IL-10 were found in patients with PsA in comparison with healthy volunteers. A statistically significant correlation was found between levels of sIL-2R and PASI, whereas no association was found with clinical parameters of joint severity. Levels of IL-1ra correlated with the number of tender and swollen joints. No correlation was found between levels of IL-6, IL-10, and clinical parameters of skin and joint severity. In the group of patients with PsA, serum levels of sIL-2R clearly correlated with severity of skin disease, whereas levels of IL-1ra were associated with joint severity. Received: 18 June 1999 / Accepted: 1 October 1999     

The effect of anti-IL-10 on proliferation and cytokine production in human schistosomiasis: fresh versus cryopreserved cells

In this study we neutralized endogenous IL-10 in PBMC from individuals chronically infected with Schistosoma haematobium by using anti-IL-10 MoAbs, and examined the effect and adult worm antigen (AWA)-specific responses in both fresh or cryopreserved cells. Anti-IL-10 alone increased background proliferation of PBMC, but did not augment the AWA-specific responses in either fresh or frozen cells. In freshly isolated PBMC, IFN-γ production in response to AWA was enhanced significantly in the presence of anti-IL-10. In cryopreserved cells, the augmentation of IFN-γ in the presence of anti-IL-10 was four-fold less than in the freshly-isolated cells. Neutralization of IL-10 had no effect on IL-4 production. These data show that IL-10 plays a role in specifically down-regulating Th1- but not Th2-type responses and, in contrast to previous reports, anti-IL-10 does not augment proliferation to parasite antigen in chronic schistosomiasis .     

Human Schistosomiasis mansoni: IL-10 modulates the in vitro granuloma formation

Granuloma formation and modulation around Schistosoma mansoni eggs that are trapped in host tissues play a pivotal role during schistosomiasis. It has been demonstrated that the granuloma reactions differ in patients with the different clinical forms of the disease. The pathology during murine schistosomiasis has been correlated with a Th2 response while resistance to infection with a Th1 type response. In humans, very little is known about the role of different cytokines on the development of the disease. Here we demonstrate that IL-10 is an important cytokine regulating the in vitro granulomatous reactivity of PBMC from intestinal (INT) patients. This was evidenced by the fact that blockage of this cytokine in the in vitro granuloma assay lead to a significant increase in granuloma size with cells from INT patients but not with individuals in the acute phase or with the hepatosplenic (HS) form of schistosomiasis. These results demonstrate for the first time that, in context with the model, a Th2 cytokine in human schistosomiasis plays an important role in controlling morbidity .     

Diagnostic potential of interleukin-40 (IL-40) in rheumatoid arthritis patients

BackgroundRheumatoid arthritis (RA) is the most common type of inflammatory arthritis. Newly emerging evidence has highlighted the role of B-cell produced cytokines in the development and progression of RA. Specifically, the expression of IL-40 has been shown to be significantly increased in affected patients.Patients and methodsThe study included 66 patients attending Rheumatology Unit, Baghdad Teaching Hospital and 66 matched controls. C-reactive protein (CRP), rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP) were measured. Disease activity score (DAS28) was assessed. Serum IL-40 levels were assessed using an enzyme-linked immunosorbent assay (ELISA).ResultsThe mean ages of patients were 46.5 ± 10.2 years (23–68 years) and were 55 females and 11 males (F:M 5:1). 4 were smokers. Positivity of CRP, RF and anti-CCP were 75.8 %, 65.2 % and 62.2 % respectively. Steroids were received by 21.2 %, methotrexate by 56.1 %, biologics in the form of etanercept and adalimumab by 86.4 %. IL-40 was significantly higher in patients (9.1 ± 1.3 ng/ml) than in controls (7.5 ± 2.2 ng/ml; p < 0.001). The IL-40 level was comparable between females and males and was not related to smoking, CRP, RF or anti-CCP positivity or to receiving medications. Serum IL-40 showed good validity in diagnosing RA at a cut-off value (≥8.2 ng/ml) and area under curve (AUC) (0.74), the sensitivity was 73.2 %, specificity (66.7 %), and the accuracy (70.5 %)(p < 0.001).ConclusionThe increased level of serum IL-40 and its potential diagnostic role have been revealed in RA. Further studies are needed to be implement and elucidate the complete role of IL-40 in RA.     

Differential effects of IL-6 blockade tocilizumab and TNF inhibitors on angiogenesis in synovial tissues from patients with rheumatoid arthritis

Objective: To compare the influences of tocilizumab (TCZ) and TNF inhibitors (TNFi) on the angiogenesis in synovial tissues of rheumatoid arthritis (RA).

Methods: Synovial tissues were obtained during joint operations from 13 RA patients treated with TCZ for at least 4 months with or without previous use of TNFi, from 13 RA patients with TNFi alone and from 10 RA patients with only conventional synthetic DMARDs (csDMARDs). Synovial tissues were evaluated by hematoxylin and eosin stain as well as by immunohistological staining with anti-CD31 in which the microvessel densities (MVD) were quantitated. Synovial histopathology was scored for various components.

Results: The most remarkable change in the synovium with TCZ was reduced angiogenesis as well as degeneration of lining layers irrespective of the previous use of TNFi. Thus, MVD in patients treated with TCZ with or without previous TNFi were significantly decreased compared with those in patients with TNFi alone or with csDMARDs. Moreover, MVD was significantly correlated with lining layer proliferation, but not with synovial stromal proliferation or inflammatory changes.

Conclusions: These results demonstrated that inhibition of angiogenesis is a unique action of TCZ. Moreover, the data also suggest that lining layers proliferation might be closely associated with angiogenesis.     

Clinical significance of serum B cell chemokine (CXCL13) in early rheumatoid arthritis patients

Background

The diagnosis of early rheumatoid arthritis (RA) is challenging. B-cell chemokine (CXCL13) plays a critical role in the disease pathogenesis.

携带hIL-4逆转录病毒重组体的构建及在类风湿关节炎中体外表达的研究

目的 构建携带人白介素4（hIL-4）表达序列的重组逆转录病毒pLXSN—hIL-4并检测目的基因的表达。方法 2004年6月至12月在中国医科大学基础医学院设计带有酶切位点的引物，含有目的基因的DNA进行聚合酶链反应扩增并纯化目的基因片段。pLXSN与目的基因片段hIL-4进行定向克隆连接，筛出阳性克隆并进行鉴定。扩增重组逆转录病毒载体pLXSN—hIL-4，并转染体外培养的人滑膜成纤维样细胞。Western blotting法测定目的基因蛋白表达水平。结果 成功构建了携带治疗基因的逆转录病毒重组体：rRV—hIL-4。Western blotting法检测到了hIL-4的表达。结论 成功构建了携带治疗基因的逆转录病毒重组体：rRv—hIL-4。以逆转录病毒为载体可以将hIL-4基因导入体外培养的人滑膜成纤维样细胞，转染后的细胞可以表达hIL-4蛋白。     

携带hIL-4逆转录病毒重组体的构建及在类风湿关节炎中体外表达的研究

目的构建携带人白介素4(hIL-4)表达序列的重组逆转录病毒pLXSN-hIL-4并检测目的基因的表达。方法2004年6月至12月在中国医科大学基础医学院设计带有酶切位点的引物,含有目的基因的DNA进行聚合酶链反应扩增并纯化目的基因片段。pLXSN与目的基因片段hIL-4进行定向克隆连接,筛出阳性克隆并进行鉴定。扩增重组逆转录病毒载体pLXSN-hIL-4,并转染体外培养的人滑膜成纤维样细胞。Westernblotting法测定目的基因蛋白表达水平。结果成功构建了携带治疗基因的逆转录病毒重组体:rRV-hIL-4。Westernblotting法检测到了hIL-4的表达。结论成功构建了携带治疗基因的逆转录病毒重组体:rRV-hIL-4。以逆转录病毒为载体可以将hIL-4基因导入体外培养的人滑膜成纤维样细胞,转染后的细胞可以表达hIL-4蛋白。     

Differential effects of interleukins 10 and 4 on the production of interleukin-6 by blood and synovium monocytes in rheumatoid arthritis

Objective. To determine how the antiinflammatory cytokines interleukin-10 (IL-10) and IL-4 affect the production of IL-6 in rheumatoid arthritis (RA) and to assess the contribution of IL-10 production. Methods. IL-6 production was measured by enzyme-linked immunosorbent assay (ELISA) in the supernatants of cultured RA synovium pieces (from 23 patients), purified RA synovial tissue monocyte/macrophages, and RA blood monocytes, in the presence of IL-10 and IL-4. IL-10 was also detected by ELISA in culture supernatants and in RA sera. Results. The production of IL-6 by RA synovium was strongly inhibited by IL-4 (46.6%; P = 0.0001) and was inhibited to a lower extent by IL-10 (25.3%; P = 0.03). Likewise, the spontaneous production of IL-6 by RA synovial tissue monocyte/macrophages was decreased by the addition of IL-4 (48.8%) and IL-10 (23.7%). This inhibition of IL-6 production was significantly lower (P < 0.03) than that observed with RA blood monocytes (83.0% for IL-10 and 85.2% for IL-4). Interestingly, and in contrast to RA blood monocytes, RA synovial tissue monocyte/macrophages produced spontaneously high levels of IL-10, which were inhibited by IL-4 and interferon-γ. Conclusion. The ability of IL-10 and IL-4 to suppress IL-6 production was dependent on 1) differences in the state of differentiation of blood and synovial tissue monocytes, and 2) local production of cytokine inside the synovium.     

Th-17 cells and serum IL-17 in rheumatoid arthritis patients: Correlation with disease activity and severity

Aim of the workTo investigate the role of T-helper 17 (Th17) cells in peripheral blood and serum interleukin-17 (IL-17) in rheumatoid arthritis (RA) patients, and their correlation with disease activity and joint destruction.Patients and methodsThis study included forty RA patients and twenty matched healthy controls. Disease activity score in 28 joints (DAS-28), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), anti-CCP, tumor necrosis factor alpha (TNF-α), serum IL-17 and Th17 cells in peripheral blood were measured. Radiological assessment using modified Sharp/van der Heijde (mSvH) score for hand and feet in addition to MRI score for the wrist and metacarpophalangeal (MCP) joints were performed for detection of synovitis and bone erosion.ResultsThe patients were 38 females and 2 males with a mean of 41.15 ± 5.85 years and disease duration of 15.6 ± 4.62 years. Serum IL-17 and Th17 cells in peripheral blood were found to be significantly increased in RA patients (204.1 ± 33.8 pg/ml and 4.62 ± 1.13%) than in controls (25.36 ± 5.39 pg/ml and 0.7 ± 0.021%) (p < 0.001). Th17 cells significantly correlated with serum IL-17 (r 0.88, p < 0.001). Both Th17 cells and serum IL-17 significantly correlated with DAS-28, ESR, CRP, TNF-α, Van der Heijde modification score and MRI scores for wrist and MCP joints for synovitis and bone erosion (all with a p < 0.001).ConclusionThis study demonstrates an important role for Th-17 cells and serum IL-17 in the pathogenesis of the inflammatory and destructive pattern characteristic of RA.     

Predictive significance of CXCL8, CXCL10 and CXCL16 in juvenile idiopathic and rheumatoid arthritis Iraqi patients

Aim of the workTo evaluate the predicative significance of three chemokines (CXCL8, CXCL10 and CXCL16) in juvenile idiopathic arthritis (JIA) and rheumatoid arthritis (RA) and to focus on their relation to some laboratory findings and clinical features.Patients and methodsSerum level of the chemokines was determined in 79 JIA and 77 RA Iraqi patients, as well as their matching controls by enzyme linked immunosorbent assay.ResultsJIA and RA patients shared significant increase of CXCL8 (24 vs. 18 and 37 vs. 15 pg/ml) and CXCL10 (38.5 vs. 17.1 and 41.5 vs. 13.2 pg/ml, respectively) compared to their controls, while no such variation observed in CXCL16 level. Regression analysis revealed that both CXCL8 and CXCL10 were significant risk factors in JIA and RA. Only rheumatoid factor-seropositive RA patients had a significantly higher CXCL10 (43 vs. 32; p = 0.014) and CXCL16 (21.2 vs. 17.7; p = 0.003) level compared seronegative patients. The CXCL10 at a cut-off value of 29.2 pg/ml in JIA showed a sensitivity of 91.1% and specificity of 91.8% and at 20.1 pg/ml in RA reached 100% and 96.2%, respectively. CXCL8 in JIA showed a sensitivity of 74.7% and specificity of 72.6% at18.6 pg/ml and in RA were 93.5% and 79.7%, respectively at 21.2 pg/ml.ConclusionsCXCL8 and CXCL10 are potential predictors of JIA and RA. CXCL10 is a more significant predictor. The CXCL16 was not found to have such impact and it might be of value in a subgroup of seropositive RA patients.