The cytotoxicity of leukocytes and lymphocytes from patients with rheumatoid arthritis for synovial cells.

Unseparated peripheral blood leukocytes obtained from patients with rheumatoid arthritis (RA) were cytotoxic for synovial cells. The cytotoxic reactions produced by RA leukocytes were more frequent and of greater magnitude than cytotoxicity induced by leukocytes from normal persons and patients with other diseases, primarily connective tissue diseases. Furthermore, the cytotoxic activity of RA leukocytes was greater for RA synovial cells than for nonrheumatoid synovial cells, in contrast to the cytotoxicity of other leukocytes, which did not discriminate between synovial cells according to their origin. Tests with purified lymphocytes showed that the cytotoxicity of unseparated leukocytes directed against RA synovial cells was due to lymphocyte cytotoxicity. These data are consistent with the possibility that sensitized lymphocytes from patients with RA recognize a distinctive antigen present on rheumatoid synovial cells.     

Dipeptidyl peptidase-IV in synovial fluid and in synovial fluid mononuclear cells of patients with rheumatoid arthritis

BackgroundDipeptidyl peptidase-IV (DPP-IV) enzymatic activity controls biological halftime of multiple local mediators. Its deregulation is associated with pathogenesis of several autoimmune diseases, including rheumatoid arthritis (RA). Although DPP-IV is the canonical representative of the group, a number of other proteins have been shown to have similar enzymatic activity. This study was aimed to identify the molecular source of DPP-IV activity in synovial fluid (SF) and fluid mononuclear cells (FMNC) in patients with RA and osteoarthritis (OA). In addition, the association of DPP-IV and the concentration of stromal cell-derived factor-1α (SDF), DPP-IV substrate, were evaluated.MethodsDPP-IV activity was measured by the kinetic fluorimetric method. The expression of studied molecules in FMNC and their concentrations in SF were assayed using flow cytometry and ELISA respectively.ResultsDPP-IV activity in SF, dominantly derived from the canonical DPP-IV, does not significantly differ between RA and OA. However, a significantly lower DPP-IV activity and expression in FMNC was found in RA as opposed to OA patients. Negative correlation between SDF concentration in SF and the relative amount of CD3+CD26+ cells was observed.ConclusionsWe report decreased presence of DPP-IV/CD26 in CD3+ FMNC in RA, which also may participate on impaired balance of SDF concentration in SF.     

Production of nitric oxide in the synovial membrane of rheumatoid and osteoarthritis patients

We have demonstrated spontaneous nitric oxide (NO) production by primary synovial cultures from rheumatoid (RA) and osteoarthritis patients. Increased NO production followed addition of staphylococcal enterotoxin B. Immunochemical double staining with specific anti-human inducible NO synthase (iNOS) and nonspecific esterase (NSE), or anti- CD68 (markers for tissue macrophages) showed that although many lining layer cells in RA synovium expressed iNOS, most (approximately 90%) were NSE- and CD68-, with only a minor population (approximately 10%) which were iNOS+, CD68+/NSE+. These data demonstrate the capacity for high output of NO by human synovial tissue and show that, although human macrophages can express high levels of iNOS, the majority of cells expressing iNOS are fibroblasts. We also report that synoviocytes, and macrophage cell lines, cultured with the NO donor, S- nitroso-acetyl penicillamine, produced high concentrations of tumor necrosis factor (TNF)-alpha. These results suggest that NO may mediate pathology in RA through the induction of TNF-alpha production.     

Nucleosides and bases in synovial fluid from patients with rheumatoid arthritis and osteoarthritis

1. Nucleosides and bases in physiological fluids result from metabolism of nucleic acids and nucleotides and from dietary sources. As nucleotide catabolism increases during tissue injury, nucleosides and bases could serve as useful biochemical markers in arthritis. 2. We have quantified nucleosides and bases in synovial fluid and plasma by high-performance liquid chromatography in order to examine whether nucleotide metabolism is increased in patients with rheumatoid arthritis and osteoarthritis. 3. At least ten u.v.-absorbing compounds were detected in plasma and synovial fluid; only urate, creatinine, hypoxanthine and uridine were present in identifiable and quantifiable amounts. 4. In synovial fluid from patients with rheumatoid arthritis the concentration of hypoxanthine was increased and that of urate decreased compared with osteoarthritis. 5. These data suggest that there is an increase in purine metabolism in the rheumatoid arthritis joint and that hypoxanthine is a potential marker of synovitis.     

Stimulation of neutrophils by insoluble immunoglobulin aggregates from synovial fluid of patients with rheumatoid arthritis

Insoluble immunoglobulin aggregates present in the synovial fluid of patients with rheumatoid arthritis have been examined for their ability to activate reactive oxidant and granule enzyme secretion from bloodstream neutrophils. These insoluble complexes activated luminol chemiluminescence, but did not activate O2-, H2O2 or granule enzyme secretion and did not activate lucigenin chemiluminescence, which also measures reactive oxidant secretion. Hence, the luminol chemiluminescence detected after activation by insoluble immunoglobulin aggregates must be due to intracellularly generated reactive oxidants, i.e. produced within phagolysosomes. Because reactive oxidant and granule enzyme secretion has occurred within rheumatoid joints, other mechanisms of neutrophil activation must exist.     

类风湿关节炎血清和滑液Th1／Th2细胞因子的水平及意义

目的初步探讨类风湿关节炎患者免疫功能紊乱的机制.方法采用酶联免疫吸附法(ELISA)检测类风湿关节炎患者血清和关节滑膜液中Th1细胞分泌的细胞因子IL-2、IFN-γ和Th2细胞分泌的细胞因子IL-6、IL-10的水平.结果类风湿关节炎患者血清及滑膜液中IL-2、IFN-γ的水平与正常对照组相比明显升高,差异具有显著性(p＜0.01或p＜0.05),而IL-6、IL-10的水平降低,差异具有显著性(p＜0.01或p＜0.05).结论在类风湿关节炎患者血清及滑膜液中Th1/Th2细胞分泌的细胞因子水平存在着明显失衡,两者以Th1细胞分泌细胞因子占优势,这可能与其发病机制密切相关.     

Elevated soluble CD8 in the synovial fluid from patients with rheumatoid arthritis

Suppressor/cytotoxic T cells express the surface marker CD8, which can be measured in a soluble form in culture supernatants of activated human lymphocytes. Using a sandwich immunoassay, we assessed the levels of soluble CD8 (sCD8) in serum from patients with rheumatoid arthritis (RA; n = 82), patients with degenerative joint disease (DJD; n = 40), and healthy controls. There were no differences in serum sCD8 levels among these groups. In contrast, the levels of soluble CD8 in the synovial fluid (SF) from patients with RA (n = 53) were significantly increased compared with the levels in 23 samples from patients with DJD (821 +/- 110 U/ml versus 213 +/- 13 U/ml, p less than 0.001). Synovial fluid sCD8 levels in the RA group were strikingly elevated, to a maximum value of 5,026 U/ml. In the majority of RA SF specimens (39 of 53), the values were significantly higher in the SF than the serum. Although the RA group had higher values of sCD8, such values were not significantly correlated with measured laboratory or clinical parameters. Current clinical and laboratory methods of evaluating patients may not be adequate in dealing with the complexity and heterogeneity of RA. Soluble CD8 values may be useful in further grouping patients with this disease.     

Identification of immunostimulatory dendritic cells in the synovial effusions of patients with rheumatoid arthritis.

Dendritic cells in the circulation are leukocytes that are rich in Ia antigens and that actively stimulate T cell replication. We have identified dendritic cells in the joint effusions of patients with rheumatoid arthritis. By phase-contrast and immunofluorescence microscopy, synovial mononuclear cells contained 1-5% dendritic profiles that were rich in HLA-DR and DQ, had small amounts of C3bi receptor, and lacked a battery of monocyte and lymphocyte markers. These dendritic cells could be enriched to 60-80% purity by cytolytic depletion of monocytes and lymphocytes with a group of monoclonal antibodies (MAb) and complement. By transmission electron microscopy, the dendritic cell processes were bulbous in shape and lacked organelles. The cytoplasm had few lysosomes or endocytic vacuoles but contained a well-developed smooth reticulum that was comparable to that previously described in the Ia-rich interdigitating cells of lymphoid tissues. The growth of sodium periodate-modified T lymphocytes was used as a rapid quantitative assay of accessory cell function. Synovial mononuclear cells were some ten times more active than normal blood cells. Treatment with alpha-Ia MAb and complement ablated stimulatory function. In contrast, removal of monocytes (MAb, 3C10) or monocytes and B (MAb, BA-1) plus T (MAb, OKT3, or T101) lymphocytes did not significantly alter total activity, and the function per viable cell increased four- to eightfold. We conclude that rheumatoid arthritis synovial fluids contain cells that are comparable in function, phenotype, and structure to blood dendritic cells, although the frequency (1-5%) is 10 times greater in joints. The reason for their accumulation in the articular cavity is not known, but dendritic cells may be important in perpetuating the joint inflammation characteristic of this disease.     

An assay for estimating the cytotoxicity of synovial fluid from patients with rheumatoid arthritis

The joint is the prime site of involvement in rheumatoid arthritis: the synovium showing lymphocytic involvement and damage. The synovial fluid shows evidence of inflammation such as increased neutrophils and raised CRP. Application of a cell-free extract of serum or synovial fluid from patients with rheumatoid arthritis, or serum from normal individuals on to 111indium oxine radiolabelled endothelial cells and fibroblasts, promotes the release of the isotope as a result of cell damage and death (cytotoxicity). In paired samples, synovial fluid was consistently more cytotoxic than rheumatoid serum. There was no difference in the cytotoxicity of rheumatoid serum or normal serum. These results demonstrate the presence of soluble factors in synovial fluid, which are capable of destroying cells, typical of those found in the synovial cavity, in blood vessels and in connective tissue generally. This system may prove to be a useful model of inflammatory damage.     

类风湿关节炎患者关节腔积液特征分析

目的 了解类风湿关节炎(RA)患者关节腔积液的特征,为该病的诊断与治疗提供实验室依据.方法 回顾分析该院2016年1月至2020年12月170例明确诊断为RA患者的关节腔积液的一般性状、细胞学检查、生化、免疫指标,并比较患者关节腔积液与血清总蛋白、尿酸、类风湿因子水平,以及不同抗环瓜氨酸肽(CCP)抗体患者的关节腔积液...     

Proto-oncogene expression in cultured synovial fibroblasts of patients with rheumatoid arthritis

Total RNA was isolated from cultured synovial fibroblasts of nine patients with rheumatoid arthritis and two controls (cruciate ligament ruptures). RNA was dot-blotted and hybridized with nine different, cloned cellular or viral oncogene probes. None of the proto-oncogenes showed a significant difference of expression in cultured fibroblasts from patients with rheumatoid arthritis when compared to the expression of control fibroblasts.     

沙利度胺对类风湿关节炎滑膜成纤维样细胞影响的体外研究

目的研究沙利度胺(Thalidomide)对类风湿关节炎(RA)滑膜成纤维样细胞(FLS)体外培养时的增生殖分化特性的影响。方法关节镜、关节活检针取RA患者滑膜组织,分离培养和鉴定FLS,MTT法和反转录-聚合酶链反应(RT-PCR)方法观察沙利度胺对FLS细胞存活分数(SF)和c-fos、COX-2mRNA表达。结果RA-FLS体外培养呈良性增生。沙利度胺对RA-FLS的SF值的抑制作用最强(P<0.05)。生理剂量的沙利度胺干预时的浓度与FLS的SF值呈负相关。RA-FLS的c-fos、环氧化酶2(COX-2)mRNA表达率高,加入沙利度胺共孵育3天后c-fos和COX-2mRNA的表达率均明显下降。结论培养和鉴定RA滑膜细胞,证实体外培养传代的RA-FLS为非恶性无限制增生。实验剂量下沙利度胺通过抑制c-fos、COX-2的表达、降低FLS增殖能力等不同途径,发挥对RA滑膜细胞的免疫调节作用。     

Collagenase-like (CL) peptidase activity in synovial fluid from patients with rheumatoid arthritis

We found the presence of collagenase-like (CL) peptidase in synovial fluid by a highly sensitive fluorescence assay using (succinyl-Gly-Pro-Leu-Gly-Pro)-4-methylcoumaryl-7-amide (Suc-GPLGP-MCA) as a substrate. Suc-GPLGP-MCA is hydrolyzed at the Leu-Gly bond by CL-peptidase. The CL-peptidase activity in synovial fluid was significantly higher in patients with rheumatoid arthritis (RA) than in patients with osteoarthritis (OA) and in arthropathy-free controls. No significant difference in CL-peptidase activity in synovial fluid was found between patients with OA and arthropathy-free controls.     

趋化因子CCL5在类风湿关节炎患者滑液炎性细胞中的表达

目的：观察趋化因子CCL5在类风湿关节炎患者体内的产生来源，以及CCL5水平与类风湿关节炎的症状和实验室检查指标的相关性。 方法：选择2004-06／10遂宁市人民医院及川北医学院附属医院收治的类风湿关节炎患者28例，以同期收治的骨关节炎患者21例为对照。所有患者取空腹外周血，有膝关节积液的11例类风湿关节炎患者和6例骨关节炎患者同时抽取关节液。用酶联免疫吸附法测定两组患者血清及滑液中CCL5水平，并对患者外周血白细胞和滑液炎性细胞体外产生CCL5进行检测。用直线相关回归分析法分析类风湿关节炎患者血清CCL5水平与关节肿胀指数、血沉、C反应蛋白之间的相关性。 结果：49例患者进入结果分析。①CCL5水平：血清中类风湿关节炎患者明显高于骨关节炎患者[（911&;#177;388），（499&;#177;289）ng／L,P〈0．05]；滑液中类风湿关节炎患者也明显高于骨关节炎患者[（1390&;#177;187），（662&;#177;154）ng/L P〈0．011；而且类风湿关节炎患者滑液中CCL5水平明显高于血清中（P〈0．01）。②相关性分析：类风湿关节炎患者血清CCL5水平与关节肿胀指数、血沉、C反应蛋白呈正相关（r=0．3229，0．4001．0．3735．P〈0．05）。③白细胞体外产生CCL5水平：类风湿关节炎患者滑液明显高于外周血和骨关节炎患者滑液（P〈0．05）；但两组患者外周血白细胞体外产生CCL5无差异（P〉0．05）。 结论：①类风湿关节炎患者血清CCL5水平增高，且与类风湿关节炎的活动性病变（关节肿胀指数、血沉、C反应蛋白）有显著相关性，表明CCL5在类风湿关节炎的病理过程中起重要作用。②患者滑液炎性细胞是CCL5产生的主要来源。