Anti-endothelial cell antibodies in sera of patients with autoimmune diseases: comparison between ELISA and FACS analysis.

In some patients suffering from rheumatoid arthritis (RA), vasculitis is a clear clinical manifestation, mentioned as rheumatoid vasculitis (RV). Autoantibodies directed against endothelial cells (AEA) have been implicated in the pathogenesis of this disorder, and it has been suggested in a number of studies that testing for AEA should be included in diagnosing RV. To test this hypothesis, we have evaluated the presence of AEA in sera of patients suffering from various autoimmune diseases, employing an ELISA with fixed cultured endothelial cells (EC). In all the groups of patients ELISA-positive sera were present. A significant difference in percentage of positivity was found between the RA and RV group (P < 0.05). In addition, our results indicated that not only antibodies directed against antigens on the EC membrane were detected, but also antibodies directed against intracellular components like DNA, histones and cytoskeletal components. Therefore, we also tested all these patient sera on unfixed intact EC using indirect immunofluorescence followed by FACS analysis. Whereas in the total patient population 34 out of 65 patients were AEA-positive as determined in the ELISA, only seven patients were weakly positive when examined by flow cytometry. We conclude that: (i) an ELISA on fixed EC does not specifically detect AEA. A positive test result is, however, to some extent correlated with the occurrence of vasculitis, and may therefore be helpful in diagnosing this disease; (ii) FACS analysis is a more suitable method than ELISA to measure the presence of membrane-specific AEA in patient sera; (iii) specific IgG-AEA are less common in patients suffering from autoimmune disorders than was assumed previously.     

Monoclonal antibodies defining a minor and a private idiotope on human rheumatoid factors

Two mouse monoclonal antibodies (mAb A216-5 and L 49-3) with antiidiotypic activity against two human monoclonal IgM rheumatoid factors (IgM RFs) were defined. Each of these monoclonal antibodies (two mouse IgG 1 K) reacted with an idiotope located on the heavy chain of the immunizing monoclonal IgM RF and was able to inhibit RF fixation to the antigen. These monoclonal antibodies did not react with other monoclonal IgM RFs from patients with macroglobulinemias or cryoglobulinemias and, therefore, did not recognize the known cross-reactive idiotopes of human monoclonal RFs. The presence of both 216-5 and 49-3 idiotopes on polyclonal IgM RFs from unrelated patients was undetectable by the inhibition assays. However, using a four-stage solid-phase radioimmunoassay, the 216-5 idiotope (minor), but not the 49-3 idiotope (private), was frequently present at a low concentration on polyclonal IgM RFs from patients suffering from rheumatoid arthritis, primary Sjögren syndromes, various infectious diseases, systemic vasculitis, and sarcoidosis and during aging. Interestingly, the 216-5 idiotope was undetectable among polyclonal IgM RFs of 12 normal adults. The main conclusions of these data are the following. (1) The definition of minor and private idiotopes of human RFs requires the use of assays able to detect low amounts of antibodies among polyclonal Ig. (2) The anti-IgG B cells which are sometimes clonally expanded during Waldenström diseases and cryoglobulinemias can also be activated during nonneoplastic diseases, among the other RF-secreting B cells. (3) Thein vivo expression of the IgM RF repertoire is different among normal adults and during various known IgM RF-inducing conditions.     

Femoral neuropathy in a patient with rheumatoid arthritis

Femoral mononeuropathy (FMN) as an extraarticular finding of rheumatoid arthritis (RA) is a phenomenon which has not been reported previously. We report a 53-year-old female patient with RA, presenting FMN findings during the course of the disease. On examination, right quadriceps and iliopsoas muscles showed grade 3 weakness on the Medical Research Council (MRC) scale. Sensory examination revealed sensory loss in the right medial leg and thigh. Patellar tendon reflex was absent in the right side. A diagnosis of a partial right femoral neuropathy was confirmed using nerve conduction study and electromyography. The probable mechanism of FMN was thought to be vasculitis.     

Synovitis score: discrimination between chronic low-grade and high-grade synovitis

AIMS: To standardize the histopathological assessment of synovial membrane specimens in order to contribute to the diagnostics of rheumatic and non-rheumatic joint diseases. METHODS AND RESULTS: Three features of chronic synovitis (enlargement of lining cell layer, cellular density of synovial stroma, leukocytic infiltrate) were semiquantitatively evaluated (from 0, absent to 3, strong) and each feature was graded separately. The sum provided the synovitis score, which was interpreted as follows: 0-1, no synovitis; 2-4, low-grade synovitis; 5-9, high-grade synovitis. Five hundred and fifty-nine synovectomy specimens were graded by two independent observers. Clinical diagnoses were osteoarthrosis (n=212), post-traumatic arthritis (n=21), rheumatoid arthritis (n=246), psoriatic arthritis (n=22), reactive arthritis (n=9), as well as controls (n=49) from autopsies of patients without joint damage. Median synovitis scores when correlated with clinical diagnoses were: controls 1.0, osteoarthritis 2.0, post-traumatic arthritis 2.0, psoriatic arthritis 3.5, reactive arthritis 5.0 and rheumatoid arthritis 5.0. The scores differed significantly between most disease groups, especially between degenerative and rheumatic diseases. A high-grade synovitis was strongly associated with rheumatic joint diseases (P<0.001, sensitivity 61.7%, specificity 96.1%). The correlation between the two observers was high (r=0.941). CONCLUSION: The proposed synovitis score is based on well-defined, reproducible histopathological criteria and may contribute to diagnosis in rheumatic and non-rheumatic joint diseases.     

类风湿关节炎的骨病变从络病论治

背景:历代医家虽对类风湿关节炎骨病变有着丰富的论述,但鲜有从络病体系探讨者。目的:从药物治疗分类、病因分析以及"络息成积"以及等几个方面进行总结报道。方法:检索中国知网、EI、SCI、ISTP数据库1900年至今的相关文献;英文检索词为"The bone erosion of Rheumatoid arthritis,TCM collaterals theory"。选取其中内外共十余篇资料从药物治疗分类、病因分析以及"络息成积"等几个方面展开分析。结果与结论:(1)类风湿关节炎骨病变诱因之一为中医之络病,内伤顽固性疾病的发展要遵从由经及络、由气入血、由功能性病变到器质性损伤的疾病发展规律;(2)类风湿关节炎骨病变之病机在于"络息成积",认为是痰聚蕴瘀,瘀存助痰,瘀积痰淫,坏血肿肉,腐骨蚀筋而使关节肿胀、畸形,甚至破坏的过程;(3)类风湿关节炎骨病变治疗之"络以通为用",目的是为了能够保证络脉的通畅,从而让脏器和气血运行正常,从而实现血管通畅,缓解类风湿关节炎病症。     

抗环瓜氨酸肽抗体检测对类风湿关节炎的临床诊断价值

目的探讨抗环瓜氨酸肽抗体（抗CCP抗体）检测对类风湿关节炎（RA）诊断的意义。方法采用酶联免疫吸附试验（ELISA）检测115份人血清的抗CCP抗体,同时采用免疫透射比浊法定量检测类风湿因子（RF）,包括40例RA患者,45例其它风湿病患者,30名正常人;并分析抗CCP抗体与RF实验结果之间的相关性。结果在40例RA病人中,抗CCP抗体的阳性率为80.0%,在其它风湿病人中的阳性率为7.0%,抗CCP抗体对RA的敏感性和特异性分别为80.0%、96.0%,其敏感性高于RF,但差异无统计学意义（P〉0.05）,特异性明显高于RF（P〈0.05）。联合应用抗CCP抗体与RF进行诊断,二者均阳性时敏感性为65.0%,特异性为97.3%。抗CCP抗体与RF实验结果之间无相关性。结论抗CCP抗体对RA具有较好的敏感性和很高的特异性,可与RF相互补充,联合检测可提高对RA早期诊断的准确性。     

Alpha-1-antitrypsin types and rheumatoid arthritis

Frequencies of alpha-1-antitrypsin (Pi) phenotypes were studied in 100 female and 100 male Swedish patients with classical rheumatoid arthritis and compared with the population frequencies. A significant increase of rare Pi types (MS, MZ, MF and SZ) was found among the patients. However, the increase concerned mainly Z heterozygotes and was more strongly pronounced in male patients. The M-subtypes showed no association with rheumatoid arthritis. Previous investigations of Pi types in rheumatoid arthritis have shown somewhat variable results. The results so far indicate, however, that an association between the Z allele and rheumatoid arthritis is likely to exist, while the evidence for a relationship between rheumatoid arthritis and other Pi alleles is considerably weaker.     

Rheumatoid arthritis with systemic necrotizing arteritis.

An autopsy case of rheumatoid arthritis with active polyarthritis, systemic necrotizing arteritis, pleuritis, pericarditis, rheumatoid nodules in a few organs and a healing gastric ulcer was reported. Histologically, systemic necrotizing arteritis was characterized by vascular changes of the following three types: Granulomatous arteritis with a characteristic arrangement of mesenchymal cells forming a palisade around coagulation necrosis of media and some of them formed a rheumatoid nodule-like lesion in the wall (RA type); Fibrinoid arteritis very similar to the Kussmaul-Maier type periarteritis nodosa (PN type); and chronic arteritis with endarterial proliferation (Ep type). Although it is hard to distinguish arteritis of PN type from the Kussmaul-Maier type periarteritis nodosa, arteritis of RA type with rheumatoid nodule-like lesion in the wall may be interpreted as an extremely developed form of vasculitis in rheumatoid arthritis.     

RHEUMATOID ARTHRITIS WITH SYSTEMIC NECROTIZING ARTERITIS

An autopsy case of rheumatoid arthritis with active polyarthritis, systemic necrotizing arteritis, pleuritis, pericarditis, rheumatoid nodules in a few organs and a healing gastric ulcer was reported. Histologically, systemic necrotizing arteritis was characterized by vascular changes of the following three types: Granulomatous arteritis with a characteristic arrangement of mesenchymal cells forming a palisade around coagulation necrosis of media and some of them formed a rheumatoid nodule-like lesion in the wall (RA type); Fibrinoid arteritis very similar to the Kussmaul-Maier type periarteritis nodosa (PN type); and chronic arteritis with endarterial proliferation (Ep type). Although it is hard to distinguish arteritis of PN type from the Kussmaul-Maier type periarteritis nodosa, arteritis of RA type with rheumatoid nodule-like lesion in the wall may be interpreted as an extremely developed form of vasculitis in rheumatoid arthritis.     

Apheresis in rheumatoid arthritis

Plasmapheresis was first introduced as a means of treating the rheumatic diseases in 1976. The rationale of its use was the removal of circulating immune complexes, thus preventing their deposition in the tissue. In rheumatoid arthritis circulating immune complexes do indeed occur and are responsible for some of the serious complications such as vasculitis. Since the T cells are implicated in tissue damage, lymphapheresis and lymphoplasmapheresis were introduced with various success rates. Controlled trials in rheumatoid arthritis have found no value for intensive plasmapheresis, limited value for lymphapheresis and possible, though suspect, value for lymphoplasmapheresis.     

Progress in hematopoietic stem cell transplantation for autoimmune diseases

An international co-ordinated Phase I/II program commenced 8 years ago to study the role of profound immunoablation with hematopoietic stem cell transplantation in the treatment of severe, refractory autoimmune disease. Almost 700 patients have been treated for a variety of autoimmune diseases, mostly multiple sclerosis, systemic sclerosis, also referred to as scleroderma, systemic lupus erythematosis, rheumatoid arthritis and juvenile idiopathic arthritis. An overall treatment-related mortality of 7% was observed, with significant differences between diseases; 11% in systemic lupus erythematosis and only one patient with rheumatoid arthritis. Although outcomes are disparate in different diseases, there were significant durable, clinically useful remissions, relapses, and nonresponders in all groups. Although different protocols were employed, a clear advantage from the more intensive myeloablative regimens was not observed, although an increased toxicity did occur. The Phase I/II data was exploited in designing the Phase III randomized, comparative trials that are running in systemic sclerosis, multiple sclerosis and rheumatoid arthritis in Europe, and at the advanced planning stage in systemic sclerosis, multiple sclerosis and systemic lupus erythematosis in the USA. In parallel, a basic science program is proceeding with the prospective studies to improve understanding of the mechanisms of autoimmune disease activity and remission.     

类风湿关节炎脂肪酸代谢相关基因的生物信息学鉴定与验证

背景:研究表明,脂肪酸代谢基因与类风湿关节炎发展紧密相关,因此基于脂肪酸代谢基因探索类风湿关节炎发病进展具有重要的临床意义。目的:探究脂肪酸代谢基因是否可以作为预测类风湿关节炎进展的可靠生物标志物。方法:从基因表达综合数据库(GEO)下载与滑膜组织相关的基因数据,应用STRING构建蛋白质-蛋白质相互作用网络分析,对其使用Cytoscape进行生物学注释(GO基因本体论)和信号通路富集分析(KEGG京都基因与基因组百科全书)。从分子特征数据库(MSigDB)筛选脂肪酸代谢相关基因,使用套索算法和支持向量机的递归特征消除算法筛选潜在生物标志物。通过CIBERSORT算法评估正常人和类风湿关节炎患者的免疫细胞浸润水平。最后,在GSE77298使用受试者工作特征曲线验证脂肪酸代谢相关基因的表达水平。结果与结论:①确定了361个类风湿关节炎差异表达基因,其中13个与报告的脂肪酸代谢相关基因重叠;②基于机器学习算法筛选出5个基因,受试者工作特征曲线显示有5个基因(PCK1、PDK1、PTGS2、PLA2G2D、DPEP2)可以预测类风湿关节炎的发展;③CIBERSORT算法结果表明上述5个基因和活化肥大细胞、中性粒细胞、静息肥大细胞、记忆性静息CD4^(+)T细胞浸润水平密切相关;④受试者工作特征曲线显示,PLA2G2D和PCK1具有较高的诊断价值;⑤提示脂肪酸代谢相关基因表达特征可作为预测类风湿关节炎临床结果的潜在生物标志物,可进一步提高类风湿关节炎预测的准确性。     

IgG rheumatoid factor, complement and immune complexes in rheumatoid synovitis and vasculitis: comparative and serial studies during cytotoxic therapy.

IgG and IgM rheumatoid factors (IgG-RF and IgM-RF), complement and three assays for immune complexes were measured in 22 patients with rheumatoid arthritis (RA) complicated by either chronic active synovitis or vasculitis. Patients with vasculitis had relatively inactive arthritis but had higher titres of rheumatoid factors, especially IgG-RF, anticomplementary activity (ACA) and lower levels of C4 than those with synovitis. Clq-binding and platelet aggregation (PA) levels were similar in both groups. Serial measurements during cytotoxic therapy showed a close temporal relationship between the clinical features of vasculitis and levels of IgG-RF, ACA and C4 both with remission and with relapse. We suggest that immune complexes containing IgG-RF which activate complement and are detected by ACA are useful markers of rheumatoid vasculitis and may be important in its pathogenesis.     

Ulcerating rheumatoid nodule of the vulva.

A case of an ulcerating rheumatoid nodule of the vulva in a 76 year old woman with rheumatoid arthritis complicated by Felty's syndrome is reported. The patient presented with a mass in the vulval region. On clinical examination, she had an ulcerated mass associated with inguinal lymphadenopathy. These findings resulted in a clinical diagnosis of invasive carcinoma of the vulva and an excision biopsy was carried out. On microscopic examination, the lesion showed the characteristic features of a rheumatoid nodule with ulceration of overlying epidermis. Adjacent vessels showed inflammation and fibrinoid necrosis of their walls suggestive of a vasculitis. Awareness of the possibility of ulceration in rheumatoid nodules may facilitate diagnosis and avert unduly aggressive treatment.     

Outcome measures in rheumatoid arthritis: the OMERACT process

The development of valid outcome measures is essential for appropriate management of any condition, but particularly chronic rheumatic diseases, such as rheumatoid arthritis. Over the last 15 years, Outcome Measures in Arthritis Clinical Trials has been dedicated to developing such measures in a variety of musculoskeletal conditions, including rheumatoid arthritis, osteoarthritis, osteoporosis, ankylosing spondylitis and gout.     

几种基因的单核苷酸多态性与类风湿关节炎

单核苷酸多态性(SNPs)是新一代的基因分子标记,随着人类基因组图谱的绘制成功,SNPs被应用于寻找各种致病基因。类风湿关节炎(RA)是一种全身自身免疫性疾病,到目前为止,其发病机制尚未完全清楚。本文简要介绍了SNPs及几种基因的SNPs与RA的关系。     

Hashimoto’s thyroiditis with granulomas: A unifying immunological etiology?

Two cases of granulomatous inflammation of the thyroid gland associated with Hashimoto’s thyroiditis are presented. In neither case is there an obvious cause of granuloma formation, the only accompanying abnormality being rheumatoid arthritis in one of the patients. Autoimmune thyroid disease has been reported in association with sarcoidosis as well as rheumatoid arthritis, diseases in which cellular immunity is activated. Immune mechanisms alone are capable of initiating and amplifying granulomatous inflammation. In this report, we suggest that the granulomas in both cases may have their origin in immunological malfunction, the same immunological malfunction responsible for Hashimoto’s thyroiditis.     

The role of KIR2DL3/HLA-C*0802 in Brazilian patients with rheumatoid vasculitis

OBJECTIVES:

Rheumatoid arthritis is a polygenically controlled systemic autoimmune disease. Rheumatoid vasculitis is an important extra-articular phenotype of rheumatoid arthritis that can result in deep cutaneous ulcers. The objective of this study was to establish a correlation between the frequency of major histocompatibility complex class I/II alleles and killer immunoglobulin-like receptor genotypes in patients with cutaneous rheumatoid vasculitis.

METHODS:

Using the Scott & Bacon 1984 criteria to diagnose rheumatoid vasculitis and after excluding any other causes such as diabetes, atherosclerosis, adverse drug reactions, infection, and smoking, patients who met the criteria were selected. All of the selected rheumatoid vasculitis patients presented deep cutaneous ulcers. Identification of the major histocompatibility complex class I/II and killer immunoglobulin-like receptor genotypes was performed by polymerase chain reaction assays of samples collected from the 23 rheumatoid vasculitis patients as well as from 80 controls (40 non-rheumatoid vasculitis RA control patients and 40 healthy volunteers).

RESULTS:

An association between the presence of the HLA-DRB1*1402 and HLA-DRB1*0101 alleles and cutaneous lesions in rheumatoid vasculitis patients and a correlation between the inhibitor KIR2DL3 and the HLA-C*0802 ligand in rheumatoid vasculitis patients were found.

CONCLUSION:

An association was found between the presence of the HLA-DRB1*1402 and HLA-DRB1*0101 alleles and the development of cutaneous lesions in rheumatoid vasculitis patients. Additionally, the HLA-C*0802 ligand protects these individuals from developing cutaneous lesions.     

Increased expression of soluble form of vascular cell adhesion molecule-1 aggravates autoimmune arthritis in MRL-Fas(lpr) mice

Vascular cell adhesion molecule-1 (VCAM-1, CD106) is important in leukocyte trafficking and its increased expression is associated with a number of chronic inflammatory diseases, including rheumatoid arthritis (RA). A soluble form of VCAM-1 (sVCAM-1) is generated by shedding of the membrane-bound molecule. The concentration of sVCAM-1 is increased in the sera of RA patients, but its pathological role has not been elucidated. The effect of sVCAM-1 relative to protection or aggravation of disease on the development of spontaneous arthritis was examined in an animal model of RA, namely MRL-Fas(lpr) mice (which display a disease resembling human RA), by generation of sVCAM-1 transgenic MRL-Fas(lpr) mice. Transgenic MRL-Fas(lpr) mice that expressed sVCAM-1 had higher incidence and increased severity of arthritis associated with higher levels of serum IgG rheumatoid factor compared with non-transgenic MRL-Fas(lpr) mice. These results suggest that sVCAM-1 plays an arthritogenic role in the development of inflammatory arthritis in MRL-Fas(lpr) mice and may present an important target for therapeutic strategy of RA.     

Rheumatoid arthritis with diffuse pulmonary rheumatoid nodules

Rheumatoid nodules in dermal or subcutaneous tissues, while indicative of rheumatoid arthritis, are very rare. It is even less common to identify these rheumatoid nodules by biopsy as well as in autopsy materials from lung tissue. These nodules may be single or multiple, which seldom cause respiratory symptoms. Here, a patient with diffuse pulmonary rheumatoid nodules and interstitial fibrosis throughout both lungs, is described. The patient, with articular symptoms and seropositivity, exhibited a rapid clinical course and died of respiratory failure 3 months after the appearance of dyspnea. Chest radiography indicated interstitial pneumonitis with bilateral diffuse peripheral shadows. At autopsy, numerous rheumatoid nodules and interstitial fibrosis had destroyed both lungs, such that no residual normal pulmonary tissue remained. It is believed that this was an extremely rare case exhibiting large numbers of rheumatoid nodules throughout the lungs. Findings with this patient indicate that, in patients with rheumatoid arthritis, clinical interstitial pneumonitis confirmed radiologically does not exclude the existence of rheumatoid lung nodules.