Comparison of the expression profile of apoptosis-associated genes in rheumatoid arthritis and osteoarthritis

The purpose of this study was to employ microarray analysis to evaluate differential gene expression in synovial tissue samples obtained from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) to study the expression profile of apoptosis-associated genes in these tissues. Four samples were obtained from RA-affected patients and three from osteoarthritis patients. After total RNA was extracted from synovial tissue, the RNA was processed using two-cycle target labeling, followed by hybridization and scanning procedure. The GeneChip Human Genome U133 Plus 2.0 containing 900471 gene loci was used and eight genes associated with apoptosis were identified with a selected p value <0.05 and a twofold change in expression in rheumatoid samples compared to osteoarthritis tissues. Anti-apoptotic genes were generally upregulated whereas apoptotic genes were downregulated suggesting that these genes may play a role in the pathogenesis of RA. Furthermore, these genes may serve as novel therapeutic targets for the treatment of RA.     

Serum and synovial fluid histidine: a comparison in rheumatoid arthritis and osteoarthritis

Summary The serum and synovial fluid (SF) histidine, sulphydryl, and protein concentrations were compared in simultaneous samples from 84 patients with rheumatoid arthritis (RA) and a control group comprising 29 patients with osteoarthritis (OA). The SF levels of histidine were higher than the serum levels in the RA patients but significantly lower than corresponding results in patients with OA (P<0.001). The latter had levels of serum and SF histidine which were equivalent and within the normal range. Greater quantities of protein were found in the SF of the patients with RA compared with the OA group. The serum and SF sulphydryl concentrations expressed as mol/g protein were low but in equilibrium in patients with RA. However the SF sulphydryl (mol/g protein) was depressed relative to serum levels in patients with OA.     

Synovial fluid levels of anti–cyclic citrullinated peptide antibodies and IgA rheumatoid factor in rheumatoid arthritis,psoriatic arthritis,and osteoarthritis

Objective

To assess the levels of anti–cyclic citrullinated peptide (anti‐CCP) and IgA rheumatoid factor (IgA‐RF) in synovial fluids of patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), and osteoarthritis (OA).

Methods

Knee effusions of 29 patients with RA (23 women, 6 men; mean ± SD age 60 ± 15 years), 20 with PsA (6 women, 14 men; mean age 51 ± 12 years), and 19 with OA (9 women, 10 men; mean age 73 ± 11.8 years) were aspirated, tested for white blood cell (WBC) counts, centrifuged, and stored at ?20°. Sera of 22, 11, and 12 of these patients with RA, PsA, and OA, respectively, were similarly stored. IgG anti‐CCP and IgA‐RF were detected by enzyme‐linked immunosorbent assay. Erythrocyte sedimentation rate and C‐reactive protein levels were used as measures of disease activity.

Results

Mean levels of synovial fluid anti‐CCP and IgA‐RF were significantly increased in RA joint effusions compared with PsA and OA (anti‐CCP: 150 ± 134, 34 ± 29, and 24 ± 26 units, respectively [P < 0.003]; IgA‐RF: 76 ± 77, 15.7 ± 10, and 18 ± 20 units, respectively). No significant difference was noted between OA and PsA. A significant correlation was found between synovial fluid anti‐CCP and serum anti‐CCP and IgA‐RF. In patients with RA, a significant correlation was found between synovial fluid WBC counts and IgA‐RF (P = 0.03) and serum IgA‐RF (P = 0.008), but not between synovial fluid and serum anti‐CCP levels. In RA patients, C‐reactive protein correlated with serum IgA‐RF.

Conclusion

Anti‐CCP and IgA‐RF were significantly increased in synovial fluid of RA in comparison with PsA and OA patients.

     

Evaluation of interleukin-6 in rheumatoid arthritis as an activity criterion

This study evaluated interleukin-6 levels as an activity criterion in rheumatoid arthritis (RA) and compared if with other activity criteria. We evaluated 35 patients with active RA, 31 with inactive RA, and 25 patients with osteoarthritis, in addition to 28 healthy individuals. Serum interleukin-6 levels were higher in active RA patients than in those with inactive RA, or osteoarthritis and healthy individuals (P<0.001). Serum interleukin-6 levels of patients with active RA were positively correlated with the erythrocyte sedimentation rate, C-reactive protein, and ₂-globulin levels (P<0.001), but there was a negative correlation with serum albumin levels (P<0.05). We conclude that interleukin-6 can be responsible for both the most systemic manifestations of RA and for its local manifestations.     

Binucleated and multinucleated forms of plasma cells in synovia from patients with rheumatoid arthritis

A morphological examination of synovial tissue from 25 patients with rheumatoid arthritis revealed that binucleated or multinucleated plasma cells were present in all samples and absent in synovia obtained from 16 control patients. Plasma cells containing two, three of four nuclei constitutet a mean 3% of the total plasma cell population. They were aways found amongst plasma cell infiltrates and in close association with small blood vessels. Ultrastructural analysis found no evidence of cellular membranes separating the individual nuclei in binucleated or multinucleated plasma cells, suggesting that the cells did not arise from fusion. Some of these plasma cells had a diameter approaching 100 μm, and many were in intimate contact with macrophages. The demonstration of a few cells with mitotic figures within the infiltrates suggests that the maintenance of plasma cell numbers in rheumatoid synovium may depend, in part, upon their local proliferation. Received: 25 August 1997 / Accepted: 2 October 1997     

Synovial proliferation differentially affects hypoxia in the joint cavities of rheumatoid arthritis and osteoarthritis patients

This study was performed to investigate whether synovial proliferation (SP) differentially affects hypoxia in the joint cavities of rheumatoid arthritis (RA) and osteoarthritis (OA) patients. Thirty RA and 42 OA patients who underwent synovitis assessment were classified into two groups based on the presence or absence of SP, as revealed by musculoskeletal ultrasonography. Synovial fluids (SFs) from the knee joints were analyzed for interleukin (IL)-8, pO(2), and white blood cell counts and blood samples were analyzed for erythrocyte sedimentation rate (ESR). No difference was found between the OA patients with and without SP in terms of SF oxygen tension (SF pO(2)) or IL-8 level, whereas the RA patients had significantly lower SF pO(2) levels in their knee joints than did the OA patients with SP, and the RA patients had higher levels of IL-8 in their joints than did the OA patients. The counts of infiltrated immune cells in the SF and tissues were much higher for patients with RA and SP than for those with OA and SP. The ESRs were not found to be correlated with SP in OA patients but were negatively correlated with SF pO(2) levels in RA patients. We conclude that ultrasonographically detected SP in OA patients does not generate a more hypoxic SF than that found in OA patients without SP. The SFs from RA patients with SP are hypoxic, which indicates that SP may have different impacts on hypoxia in the joint cavities of RA and OA patients.     

Early lymphocyte activation in the synovial microenvironment in patients with osteoarthritis: comparison with rheumatoid arthritis patients and healthy controls

Osteoarthritis (OA) is largely considered to be a non-inflammatory disease, although there is compelling evidence that subclinical inflammation is a common event, even in the absence of acute inflammatory flares. In this study we analyze, by means of CD5 and CD69 expression, the infiltration and early activation of CD5+cells, mostly lymphocytes, in both synovial membrane and synovial fluid from advanced OA patients and compare them with samples from patients with rheumatoid arthritis and healthy controls. The number of infiltrating CD5+ cells in both synovial membrane and synovial fluid from patients with advanced OA was significantly reduced as compared with rheumatoid arthritis patients. However, synovial membrane and synovial fluid CD5+ cells on OA exhibited a phenotype with evidence of recent activation comparable to that observed in RA.     

Comparison of modern marker proteins in serum and synovial fluid in patients with advanced osteoarthrosis and rheumatoid arthritis

Numerous studies have focused on the significance of modern marker proteins in the synovial fluid of the knee joint and in the serum both, for osteoarthritis (OA) and rheumatoid arthritis (RA). The relationship between the serum concentrations and the concentrations in the synovial fluid is still unclear. Synovial fluid and serum samples were obtained from 13 patients with advanced OA and from 8 patients with severe RA and concentrations of MMP-1, MMP-3, MMP-13, TIMP-1, COMP and MIA/CD-RAP were determined. All values were normalized against the total protein concentrations. Serum concentrations of MMP-13 in the RA-group were statistically higher than the synovial values (P<0.05). MMP-13 was the only marker protein that revealed distinct higher levels in the serum than in the synovial fluid. The study design allows only conclusions about advanced stages of RA and OA. Longitudinal investigations may provide further information about the value of MMP-13 as a potential marker to monitor the course of RA and OA.     

Immunohistochemical detection of factor XIIIa and factor XIIIs in synovial membranes of patients with rheumatoid arthritis or osteoarthritis

In spite of differences in etiology, RA and OA lead to astonishingly similar synovitic alterations. Fibroblastic transformation of the synovial membrane and an increase in monocytes constitute a rare but highly characteristic feature of RA. Monocytes synthesize factor (F) XIII, implying that FXIII (a and s) in synovial tissue might help to differentiate between RA and OA. Biopsies were obtained at open surgery from 98 unselected patients with the clinical diagnosis of RA (n=54) or OA (n=44). In a three-stage (ABC) immunoperoxidase technique, polyclonal antisera against factor XIIIa and factor XIIIs were investigated. Compared to OA sections, RA synovium showed more FXIIIa-positve cells - monocytes, fibrocytes, fibroblasts and synovial lining cells. In the subsynovial layer, band-like structure of FXIIIa-stained cells was observed in 27.8% of the RA patients, but in only one OA specimen. Higher proportions of FXIIIa-positive monocytes, macrophages, histiocytes and fibroblasts, as well as positive Langhans' giant cells and vascular wall regions (except endothelial cells), were observed in RA. OA specimens revealed more intense FXIIIa labeling of these cells with a lower percentage of stained cells. Overall, labeling with FXIIIs antibody resulted in less intense staining. In conclusion, distinction between synovitis caused by RA and synovitis due to OA is possible, as the former show higher numbers of FXIIIa-positive cells, including monocytes, fibroblasts, fibrocytes and synovial lining cells. Furthermore, RA tissue is stained less intensely than OA tissue. There is evidence for continuous excretion of FXIII in the synovial membrane by the above-mentioned cell systems.     

蛋白酶激活受体2、NF-kB在溃疡性结肠炎肠黏膜的表达及其与肥大细胞相关性研究

目的探讨溃疡性结肠炎（UC）病人肠黏膜蛋白酶激活受体2（PAR-2）、核转录因子-kB（NF—kB）表达和肥大细胞的变化,以及三者在UC发病机制中的作用。方法32个黏膜标本取自行结肠镜检查的UC病人,并对UC的病理组织学炎症进行分级,Ⅰ、Ⅱ级12例,Ⅲ、Ⅳ级20例;对照组肠黏膜取白15名健康成人。半定量RT—PCR检测两组肠黏膜PAR-2的mRNA表达,免疫组化方法检测PAR-2、NF—KBp65蛋白表达和肥大细胞数量。结果与对照组相比,UC组肠黏膜中PAR-2mRNA和蛋白过度表达,PAR-2mRNA表达与疾病严重程度正相关（P〈0．01）。免疫组化结果显示,UC组肠黏膜肥大细胞数显著高于对照组（P〈0．01）正常对照组肠黏膜无或微弱阳性PAR-2、NF—KBp65表达。病理分级Ⅲ、Ⅳ级肠黏膜PAR-2、NF—KBp65蛋白表达及肥大细胞数均较病理分级Ⅰ、Ⅱ级组明显增加（P〈0．05）。PAR-2与肥大细胞数量之间呈正相关（r=0．78,P〈0．01）,PAR-2与NF—kBp65之间也呈正相关（r=0．56,P〈0．01）。结论UC的发生发展与PAR-2、NF—KBp65的过度表达及肥大细胞数量变化密切相关。     

Plasma and synovial fluid concentrations of sulphasalazine and two of its metabolites in rheumatoid arthritis

Summary A study was made of plasma and synovial fluid levels of sulphasalazine, one of its dissociation products — sulphapyridine and a metabolite of the latter — acetyl sulphapyridine in patients with rheumatoid arthritis (RA) who were in a steady state on sulphasalazine therapy. Combined sulphapyridine levels were significantly higher than those of sulphasalazine both in plasma and synovial fluid. Synovial fluid levels of both drugs correlated with their plasma levels and were generally slightly lower. Some patients accumulated sulphasalazine and sulphapyridine in the synovial fluid and the mean concentration of sulphasalazine was higher in the fluid than in the plasma. The explanation for this is uncertain. The concentration of combined sulphapyridine in synovial fluid was related to local joint inflammation and more active systemic disease. No consistent association was found between sulphasalazine levels and local or systemic activity. The higher sulphapyridine levels in synovial fluid found in this study suggest the possibility that this moiety could play a more active role in RA than it does in inflammatory bowel disease.     

肥大细胞及蛋白酶激活受体-2在大鼠实验性肝纤维化中的变化

目的观察肝纤维化大鼠的肥大细胞(MC)数量变化和蛋白酶激活受体-2(PAR-2)在肝脏中表达及其与肝纤维化的关系。方法建立大鼠肝纤维化模型,以甲苯胺蓝染色显示MC,运用碱水解法测定肝脏羟脯氨酸含量,采用逆转录聚合酶链反应(RT-PCR)方法检测正常对照组及造模2、4、8、12周组大鼠肝组织中PAR-2 mRNA的表达,免疫组织化学方法检测PAR-2蛋白的表达及定位。结果正常对照组大鼠肝组织中MC数量极少,仅(2．5±1．0)个,主要沿肝脏汇管区分布；模型组MC数量：2周为(9．1±0．5)个,4周为(15．7±3．0)个,8周为(32．0±3．3)个。造模组2周与正常组比较,P=0．038,造模组组间比较,F=58．553,P＜0．01,差异均有统计学意义。MC在汇管区、中央静脉周围密集分布。随着造模时间的延长,肝脏羟脯氨酸的含量逐渐升高。PAR-2 mRNA检测结果,PAR- 2/β-肌动蛋白(β-actin)：正常对照组几乎不表达；造模2周肝组织可有少量PAR-2 mRNA表达,为0．15±0．01,4周为0．35±0．02,8周为0．80±0．02。PAR-2蛋白检测结果,阳性信号灰度：正常对照组为156．0±0．5；造模2周为162．5±1．3,4周为174．8±1．3,8周为185．7±2．1,12周为207．7±4．4,模型组与对照组比较和模型组组间比较,F=429．389,P＜<0．01,差异均有统计学意义。结论PAR-2 mRNA和蛋白的表达水平与MC数量、羟脯氨酸含量的增加一致,可能在肝纤维化的发生发展过程中起重要作用。     

T cell subsets in the blast cell population in phytohemagglutinin-stimulated peripheral blood in vitro and in rheumatoid arthritis synovial fluid in vivo

Summary The immunomodulatory T4/T8 ratio was studied in the total and activated lymphocyte populations by a method combining visualisation of ³H-thymidine incorporating blasts with autoradiography (AR) with simultaneous identification of the respective lymphocyte subsets using monoclonal antibodies in avidin-biotin-peroxidase complex (ABC) staining. In rheumatoid arthritis synovial fluid the activated T4/T8 ratio (calculated from T cells in the S phase of the cell cycle) was significantly different from the total T4/T8 ratio (calculated for all the T cells) (0.45±0.05 versus 0.69±0.05, P<0.01). Similarly, the activated and total T4/T8 ratios were also significantly different in the phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cell cultures at days 3 and 5.     

Infiltrations of plasma cells in synovium are highly associated with synovial fluid levels of APRIL in inflamed peripheral joints of rheumatoid arthritis

Infiltration of plasma cells can be a histopathological hallmark of articular synovium with rheumatoid arthritis (RA). A proliferation-inducing ligand (APRIL) may have key roles in homeostasis and development of B cells, and the differentiation of B cells into plasma cells. This study was designed to explore the relationships between the infiltrations of plasma cells in synovium and the synovial fluid levels of APRIL in inflamed peripheral joints of RA. Synovium and synovial fluid were sampled from 21 RA patients underwent arthroscopic synovectomy for inflamed peripheral joints. The variants of rheumatoid synovium were classified into the follicular and diffuse synovitis by hematoxylin and eosin staining, and the infiltrations of plasma cells in rheumatoid synovium were quantified under the light microscope. The synovial fluid levels of APRIL were measured with the enzyme-linked immunosorbent assay. The mean number of infiltrating plasma cells in synovium and the mean synovial fluid level of APRIL were significantly increased in follicular synovitis compared with those in diffuse synovitis (P = 0.009, and P = 0.018, respectively), and there was a highly positive association between the infiltrations of plasma cells and the synovial fluid levels of APRIL among all of the RA patients (Rs = 0.776, P < 0.001). These findings suggest that the local production of APRIL may be associated with the ectopic lymphoid neogenesis in rheumatoid synovium and may have a role in contributing to the infiltration of plasma cells in synovium within inflamed peripheral joints of RA.     

Comparison of blood and synovial fluid lymphocyte subsets in rheumatoid arthritis and osteoarthritis

Summary Immunoregulatory T-cell deficiency is thought to underlie pathogenesis of rheumatoid arthritis (RA) as a systemic autoimmunopathy. The aim of this study was a simultaneous analysis of peripheral blood and synovial lymphocyte subsets (Ly-SS) of RA patients as compared to patients with locally active osteoarthritis (OA). Peripheral blood Ly-SS and paired synovial fluid Ly-SS from 87 RA patients were analysed by two dimensional flow cytometry (Simulset Becton Dickinson) as compared to 15 OA patients. The control group consisted of 32 healthy subjects. The peripheral blood analysis from RA and OA patients revealed a significant decrease of CD8+T-cells and increase of CD4+:CD8+ ratio when compared to the control group. The blood of RA patients showed a significant increase of HLA DR+ and IL 2R+T cells as compared to OA group. The synovial fluid from RA and OA patients showed a significant increase of CD3+, CD8+, HLA DR+ T-cells and decrease of CD4+:CD8+ ratio and CD19+ cells in comparison to the peripheral blood. This study shows, that the OA T-cell system seems not to be activated in peripheral blood in opposition to RA patients. Synovial fluid Ly-SS in OA, however, showed only quantitative but not qualitative differences. OA seems to be mainly a local inflammatory response depending on T-cells, when lymphocyte T activity in blood is diminished.     

肥大细胞及蛋白酶激活受体-2与肝纤维化

近年来研究表明肥大细胞与肝纤维化的发生发展有密切关系,但其具体作用途径不甚明确,目前推测肥大细胞可能通过蛋白酶激活受体-2途径在肝纤维化中发挥重要作用.现将肥大细胞、蛋白酶激活受体-2与肝纤维化的有关进展作一综述.     

Atlanto-odontoid osteoarthritis in rheumatoid arthritis: dynamic CT findings

We analyzed the CT appearances of degenerative change in the atlanto-odontoid joint (AOJ) in patients with rheumatoid arthritis (RA) and evaluated the effect of these changes on atlanto-axial joint (AAJ) rotation by dynamic CT. This revealed that 9 patients (24%) treated with methotrexate had degenerative features in the AOJ. The ratio of AAJ rotation to the total rotation of the cervical spine was significantly higher in normal subjects (54±3%) than in patients (38±12%). The degree of AAJ rotation was significantly lower in the patient group with degenerative features in the AOJ (20.9±8.4°) than in patients without degenerative features (28.5±7.4°). RA patients with a history of longstanding disease and treatment with antirheumatic drugs may develop AO OA. Although secondary OA was described as healing phenomena in the joints of RA patients, it can limit rotation in the AAJ and cause suboccipital neck pain. A regular check-up of the AAJ and AOJ by means of dynamic CT in all RA patients is proposed to avoid possible antirheumatic drug complications.Abbreviations AAJ Atlanto-axial joint - AOJ Atlanto-odontoid joint - MTX Methotrexate - OA Osteoarthritis - RA Rheumatoid arthritis     

Serum and synovial fluid leptin levels and markers of inflammation in rheumatoid arthritis

This study was designed to investigate the serum and synovial fluid leptin levels, and inflammatory markers in rheumatoid arthritis (RA) patients. Serum and synovial fluid leptin levels were significantly higher (P > 0.05) in RA patients than control group; RA patients with moderate disease activity (DAS < 2.7) having significantly higher leptin levels (P > 0.05) than those with low disease activity (DAS < 2.7). Leukocytes and erythrocyte sedimentation rate (ESR) were found to be significantly higher in moderate disease activity RA group compared to low activity group (P > 0.05, P < 0.001, respectively). Serum leptin level is found to be independent of age and inflammatory markers. ESR is positively correlated with DAS activity and CRP values. Our finding of no correlation between leptin and BMI shows that regulation of leptinemia is complex, and leptin levels cannot be used to assess RA activity.     

Tumor necrosis factor receptor 2 M196R polymorphism in rheumatoid arthritis and osteoarthritis: relationship with sTNFR2 levels and clinical features

We investigate the clinical association of tumor necrosis factor receptor 2 (TNFR2) M196R polymorphism with rheumatoid arthritis (RA) and knee osteoarthritis (OA). Acute phase reactants, lipid profile, sTNFR2 levels, disease activity–disability indexes, and TNFR2 M196R polymorphism were analyzed in 50 RA, 50 knee OA patients, and 120 healthy subjects (HS). The M/M genotype frequency was 0.74 (RA), 0.80 (OA), and 0.64 (HS). The M/R genotype frequency was RA (0.26), OA (0.20), and HS (0.29). The R/R genotype was observed only in HS (0.07). The M allele was associated with OA (P = 0.0137, OR = 2.43). Total cholesterol, triglyceride levels, apolipoprotein A-I and B showed significant differences (P < 0.05). The highest sTNFR2 levels were observed in RA and OA (P = 0.001), however M/M and M/R carriers do not correlate with sTNFR2 production. Our findings suggest an association of the M allele with knee OA. In addition, high sTNFR2 levels in RA and OA were found.