Associations between HLA-DRB1, RANK, RANKL, OPG, and IL-17 genotypes and disease severity phenotypes in Japanese patients with early rheumatoid arthritis

We examined associations between human leukocyte antigen DRB1 (HLA-DRB1) shared epitope (SE), receptor activator of nuclear factor-kappaB (RANK), RANK ligand (RANKL), osteoprotegerin (OPG), and interleukin 17 (IL-17) genotypes with age of disease onset and radiographic progression in Japanese patients with early rheumatoid arthritis (RA). HLA-DRB1 genotypes were evaluated in 123 patients with early RA (98 female, 25 male) within 1 year of symptom onset. In 72 patients, radiographic progression over a 2-year period was evaluated using Larsen’s methods, and genotypes of three polymorphic sites in RANK, five sites in RANKL, two sites in OPG, and three sites in IL-17 were determined by direct polymerase chain reaction sequencing. Possession of an SE allele was significantly associated with earlier disease onset in females (median 46.9 vs 51.9 years in SE− patients; P = 0.04). Single nucleotide polymorphisms (SNPs) in RANKL (rs2277438, P = 0.028) and IL-17 (rs3804513, P = 0.049) were significantly associated with radiographic progression at 2 years. RANKL-G−, SE− patients (n = 12) had significantly less joint damage than did RANKL-G+, SE− patients (n = 11; P = 0.0038), RANKL-G−, SE+ patients (n = 21; P = 0.0018) and RANKL-G+, SE+ patients (n = 28; P = 0.0024). In Japanese RA patients, HLA-DRB1 SE alleles are associated with disease onset at an earlier age, as has been observed in Caucasian RA patients. In addition, SNPs in RANKL and IL-17 may be associated with radiographic progression in Japanese patients with early RA.     

Frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells correlate with disease activity in rheumatoid arthritis

Abstract

Objective. Rheumatoid arthritis (RA) is a common autoimmune disease that is primarily driven by effector T cells, particularly Th17 cells, which are mainly contained within CD4+CD161+ T cells. Thus, we aimed to explore whether the frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells were correlated with RA disease activity.

Methods. The surface phenotype and cytokine production of blood were analyzed by flow cytometry in 52 RA patients and 17 healthy controls. The disease activity was evaluated by the 28-joint disease activity score.

Results. The frequencies of circulating IL-17-producing CD4+CD161+ T cells and CD4+CD161+ T cells were increased in RA patients, and they were elevated in patients with active disease status compared to patients with low disease status. Furthermore, their frequencies were positively correlated with disease activity parameters. Receiver operating characteristic curve analysis revealed that IL-17-producing CD4+CD161+ T cell levels were able to distinguish disease activity with 60.7 % sensitivity and 87.5 % specificity, while CD4+CD161+ T cell levels showed 92.9 % sensitivity and 66.7 % specificity.

Conclusion. These results support the hypothesis that Th17 cells are involved in the pathogenesis of RA and suggest that circulating CD4+CD161+ T cells are a potential biomarker of RA disease activity.     

Serum interleukin-23 level in rheumatoid arthritis patients: Relation to disease activity and severity

Aim of the work

The aim of this study was to evaluate interleukin-23 (IL-23) level in the sera of rheumatoid arthritis (RA) patients and to determine its relation with disease activity and severity.

Relation between serum IL-17 level and risk of osteoporotic fracture in premenopausal rheumatoid arthritis patients: Clinical,radiological and laboratory studies

IntroductionOsteoporosis is a main extra-articular complication of rheumatoid arthritis (RA) which may lead to fractures. Interleukin-17 (IL-17) is one of the cytokines which plays a significant role in RA pathogenesis and promotion of osteoporosis.Aim of the workTo study the relation between serum IL-17 levels and the risk of osteoporotic fractures in pre-menopausal RA patients.Patients and methodsTwenty-five premenopausal RA patients and 20 matched healthy controls were included in this study. All patients were subjected to detailed history taking, thorough clinical examination, disease activity assessment using the disease activity score-28 (DAS-28) and disability was assessed using Health Assessment Questionnaire–Disability Index (HAQ-DI). Bone mineral density and serum IL-17 levels were measured in patients and the control. Fracture Risk Assessment Tool (FRAX index) was also calculated.ResultsThe mean age of RA patients was 38.8 ± 7.6 years. The BMD was significantly reduced in patients compared to the control at the femur neck (p = 0.008), wrist (p = 0.046) and at the lumbar spine (p = 0.005). The Z score was below the expected range for age in 36% compared to 5% in the control (p = 0.03). Serum IL-17 concentrations were significantly higher in patients (5.99 ± 1.22 pg/ml) compared to the control (3.73 ± 2.15 pg/ml) (p < 0.001). Serum IL-17 levels showed a significant correlation with FRAX scores. Z-score interpretation showed a strong positive significant correlation with FRAX index; major osteoporotic fractures and hip fracture (p = 0.005 and p = 0.013, respectively) in patients.ConclusionThe premenopausal Rheumatoid arthritis patients showed a high fracture probability. Interleukin-17 serum level is associated with higher liability to fractures among rheumatoid patients.     

Serum and synovial fluid interleukin-17 concentrations in rheumatoid arthritis patients: Relation to disease activity,radiographic severity and power Doppler ultrasound

Aim of the workTo investigate serum and synovial fluid levels of IL-17 in rheumatoid arthritis (RA) patients, and its correlation with disease activity and severity.Patients and methods20 RA patients together with 20 primary knee osteoarthritis (KOA) patients and 15 healthy individuals matched for age and sex as control groups were enrolled in this study. Both RA and KOA patients presented with knee effusion. Paired samples of serum and synovial fluid (SF) were collected from RA, OA patients and serum samples from the healthy individuals. RA disease activity was assessed using DAS-28 score and power Doppler ultrasound (PDUS) according to the European League against Rheumatism (EULAR). Radiographic damage was evaluated according to Larsen score.ResultsSerum levels of IL-17 were significantly elevated in RA patients compared to controls (p < 0.001). Also, SF of IL-17 was significantly higher in RA patients compared to OA patients (p < 0.001). In addition, synovial level of IL-17 was significantly higher in RA patient compared to their serum level (p < 0.001). Regarding disease activity grading among RA patients, significant differences (p < 0.05) in mean serum and synovial IL-17 levels were reported being higher in severe active disease. Positive correlations of serum and SF IL 17 levels with PDUS findings and Larsen score were reported.ConclusionSerum and synovial IL-17 levels were significantly elevated in RA patients which clarifies its possible role in RA pathogenesis and correlates positively with disease activity parameters, PDUS findings and Larsen score. Thus targeting IL-17 may provide a promising role in suppressing RA.     

Interleukin-17 gene expression in patients with rheumatoid arthritis

Abstract

Interleukin-17 is a proinflammatory cytokine. Recent animal studies have shown that IL-17 plays a role in the initiation and progression of arthritis. However, whether IL-17 has a prominent role in human rheumatoid arthritis (RA) or not remains unclear. Here we investigated the role of IL-17 in patients with RA. cDNA was prepared from knee joint synovial tissues of RA (n = 11) and osteoarthritic (OA, n = 10) patients and PBMC of RA (n = 52) and healthy subjects (n = 34). IL-17 gene expression level was measured by real-time PCR, and was compared with various clinical parameters. IL-17 gene expression in synovial tissues of RA was similar to that in OA. IL-17 gene expression level in PBMC of RA patients was significantly higher than in the control. The response (changes in DAS) to two-week treatment with anti-TNF-α blockers (infliximab or etanercept) did not correlate with changes in IL-17 gene expression levels. The IL-17/TNF-α gene expression ratio at baseline (before treatment) tended to be lower in responders to the treatment. Expression of IL-17 gene in PBMC may be associated with the inflammatory process of RA. IL-17/TNF-α expression ratio is a potentially suitable marker of response to anti-TNF-α therapy.     

血清高迁移率族蛋白1和白细胞介素-17在类风湿关节炎患者血清中的表达及治疗前后的变化

目的 探讨类风湿关节炎(RA)患者血清高迁移率族蛋白1(HMGB1)和白细胞介素-17 (IL-17)水平及其与RA病情的关系,分析抗风湿药物的治疗作用.方法 选取61例RA患者(活动期35例,低活动度期26例)和24例健康体检者为对照组.25例随访RA活动期患者应用药物治疗后,分为缓解组(达美国风湿病学会50％改善标准,ACR50)和未缓解组(未达ACR50).采用酶联免疫吸附试验(ELISA)检测血清中HMGB1和IL-17水平.结果 活动期RA患者血清HMGB1和IL-17水平明显高于对照组和低活动度RA患者.RA患者HMGB1和IL-17水平与疾病活动性评分指标(DAS28)呈正相关,HMGB1与IL-17呈正相关.多元回归分析显示,HMGB1与DAS28存在线性依存关系.药物治疗后,HMGB1和IL-17水平在缓解组RA患者明显下降,未缓解组无显著变化.结论 HMGB1和IL-17可能参与RA的发病,并可能成为监测病情和疗效的指标之一.     

Interleukin-17 gene expression in patients with rheumatoid arthritis

Interleukin-17 is a proinflammatory cytokine. Recent animal studies have shown that IL-17 plays a role in the initiation and progression of arthritis. However, whether IL-17 has a prominent role in human rheumatoid arthritis (RA) or not remains unclear. Here we investigated the role of IL-17 in patients with RA. cDNA was prepared from knee joint synovial tissues of RA (n = 11) and osteoarthritic (OA, n = 10) patients and PBMC of RA (n = 52) and healthy subjects (n = 34). IL-17 gene expression level was measured by real-time PCR, and was compared with various clinical parameters. IL-17 gene expression in synovial tissues of RA was similar to that in OA. IL-17 gene expression level in PBMC of RA patients was significantly higher than in the control. The response (changes in DAS) to two-week treatment with anti-TNF-α blockers (infliximab or etanercept) did not correlate with changes in IL-17 gene expression levels. The IL-17/TNF-α gene expression ratio at baseline (before treatment) tended to be lower in responders to the treatment. Expression of IL-17 gene in PBMC may be associated with the inflammatory process of RA. IL-17/TNF-α expression ratio is a potentially suitable marker of response to anti-TNF-α therapy.     

IL-17-producing T cells can augment autoantibody-induced arthritis

Rheumatoid arthritis is a T lymphocyte-mediated disorder, but the precise nature of T cell involvement remains unclear. In the K/BxN mouse model of inflammatory arthritis, T cells initiate disease by providing help to B cells to produce arthritogenic autoantibodies. Here, we have characterized an additional, nonhumoral role for T cells in promoting autoantibody-induced arthritis. Autoreactive KRN T cells introduced either by direct transfer or bone marrow transplantation into B-cell-deficient hosts enhanced K/BxN serum-transferred arthritis, an effect that was dependent on expression of the cognate MHC-molecule/peptide complex. The T cell influence was dependent on interleukin (IL)-17; in contrast, standard serum-transferred arthritis, unenhanced by the addition of T cells, was unaffected by IL-17 neutralization. An IL-17-producing population of transferred KRN T cells was identified and found to be supported by the cotransfer of arthritogenic autoantibodies. IL-17-producing KRN T cells were enriched in inflamed joints of K/BxN mice, suggesting either selective recruitment or preferential differentiation. These results demonstrate the potential for autoreactive T cells to play two roles in the development of arthritis, both driving the production of pathogenic autoantibodies and bolstering the subsequent inflammatory cascade dependent on the innate immune system.     

Effects of rheumatoid factor isotypes on disease activity and severity in patients with rheumatoid arthritis: a comparative study

The value of rheumatoid factor (RF) isotypes for assessing rheumatoid arthritis (RA) remains debatable. In this study, we have examined the relationships between RF isotypes and disease activity and severity in RA patients. Sixty-two patients with RA, 48 women and 14 men, were studied. RF was measured by nephelometry (RF–N) and IgG–, IgA–, and IgM–RF isotypes were measured using enzyme-linked immunosorbent assay. Serum C-reactive protein and erythrocyte sedimentation rate were also determined. The patients were classified according to disease activity, joint damage, functional status, and presence of pulmonary involvement, rheumatoid nodule, and secondary Sjögren’s syndrome. Although the patients with active disease had significantly higher IgA–RF and IgM–RF levels compared to inactive patients, IgA–RF and IgM–RF were not found to be independently associated with disease activity in multivariate analysis. In patients with severe joint damage, IgA–RF and RF–N were significantly higher than those of the other patients. Multiple regression analysis showed that IgA–RF was the unique variable independently associated to severe joint damage. The patients with class III and IV functional index had significantly higher IgM–RF, IgA–RF, and RF–N levels compared to the patients with class I and II functional index; however, RFs were not significantly associated with functional status in multivariate analysis. IgA–RF and IgM–RF were significantly associated with pulmonary involvement and rheumatoid nodule, respectively. No significant associations were found between RF isotypes and secondary Sjögren’s syndrome. Our results suggest that the clinical usefulness of IgA and IgM isotypes is better than RF–N. Elevated IgA–RF may be a marker of erosive disease. The usefulness of RF isotypes for monitoring disease activity or functional status appears to be limited.     

Methylene tetrahydrofolate reductase (MTHFR) gene polymorphisms in rheumatoid arthritis patients: Correlation with serum osteopontin levels and disease activity

BackgroundRheumatoid arthritis (RA) is a systemic, chronic inflammatory disease with genetic predisposition. Osteopontin (OPN) is overexpressed in RA and plays a key role in the perpetuation of synovitis. Not all RA patients show the same level of response to methotrexate (MTX) suggesting genetic variations in the drug-metabolizing enzymes.Aim of the workTo detect methylene-tetra-hydrofolate reductase (MTHFR) 677C/T and 1298A/C gene polymorphisms in RA patients treated with MTX and to investigate the relationship with serum OPN levels and disease activity.Patients and methods62 RA patients and 21 healthy controls were included. Serum OPN was measured using ELISA. Genotyping of MTHFR gene was carried out by polymerase chain reaction-restriction fragment length polymorphism. Disease activity score in 28 joints (DAS28) and the modified health assessment questionnaire (MHAQ) were assessed.ResultsThe patients’ age was 42.7 ± 12.7 years, F:M (4.6:1) and a disease duration of 5.7 ± 4.6 years. Their DAS28 was 4.1 ± 1.6 and the MHAQ (median 1; range 0–2.3). Serum OPN levels in RA patients (median 8.8; range 4–44.5 ng/ml) were significantly higher than in control (5.6; 2.1–10.9) (p = 0.002). In RA patients, serum OPN significantly correlated with the duration of morning stiffness (p = 0.009), ESR (p < 0.0001) and DAS28 (p < 0.0001). MTHFR (677C>T) polymorphisms significantly correlated with MHAQ (p = 0.012) while (1298A>C) polymorphisms significantly correlated with tender joint count (p = 0.04). OPN levels were higher among patients with MTHFR (1298A/C) AC genotype (8.9; 4.1–33.9 ng/ml), while in those with (677C>T) polymorphisms it was higher among those with CT genotype (8.9; 4.1–44.5).ConclusionSerum OPN level relates with the degree of rheumatoid activity.     

白介素-17和抗-CCP抗体联合检测对类风湿关节炎活动度及骨侵蚀的预测价值

目的探讨联合检测白介素（IL）-17和抗环瓜氨酸肽（CCP）抗体对类风湿关节炎（RA）活动度及骨侵蚀的预测价值。方法收集80例RA患者和30例健康体检者（健康对照组）的血液样本,应用ELISA检测血清IL-17和抗-CCP抗体,结合双手X线分期,分析RA患者与健康对照者之间,及RA早期与非早期,RA活动期与缓解期,RA骨侵蚀与非骨侵蚀之间IL-17、抗-CCP抗体的差异。结果 （1）RA患者的IL-17、抗-CCP抗体高于健康对照者,差异有统计学意义（P〈0.05）;（2）早期RA组与非早期RA组之间比较,IL-17、抗CCP抗体水平差异无统计学意义（P〉0.05）,RA活动组高于缓解组、骨侵蚀组高于非骨侵蚀组,差异有统计学意义（P〈0.05）。结论 IL-17、抗-CCP抗体联合检测对类风湿关节炎活动度及骨侵蚀具有预测价值。     

CD14++CD16+ monocyte subset expansion in rheumatoid arthritis patients: Relation to disease activity and interleukin-17

Aim of the workMonocytes are divided into three major subsets based on the expression of the cluster of differentiation CD14 and CD16. The aim of this work was to determine which of the CD16⁺ monocyte subpopulations is expanded in rheumatoid arthritis (RA) and its association with disease activity and interleukin-17 (IL17) levels.Patients and methodsFifty-three RA patients and 20 controls were enrolled in this study. Flow cytometry was performed to detect monocyte subsets and IL17 was measured by ELISA. Disease activity score (DAS28) was assessed.ResultsCD14⁺⁺CD16⁺ monocyte percentage was significantly higher in long standing RA patients compared with early patients and controls (p < 0.01, p < 0.001 respectively). It was significantly higher in patients with RA disease activity and remission compared with the controls (p < 0.001, p < 0.01 respectively). It was not significantly associated with resistance to disease modifying antirheumatic drugs (DMARDs), C-reactive protein, rheumatoid factor and anti-CCP positivity (p > 0.05). It significantly correlated with IL17 (p < 0.002). CD14⁺CD16⁺ monocyte percentage was not significantly correlated with any of the above parameters. IL17 level was significantly higher in patients with early and long standing RA compared to controls (p < 0.01, p < 0.001 respectively). IL17 was higher in RA patients with active disease compared to those in remission and controls (p < 0.01, p < 0.001 respectively). It was higher in RA patients resistant to DMARDs than in responding patients (p < 0.017).ConclusionCD14⁺⁺CD16⁺ monocyte subpopulation was expanded in long standing RA and was correlated with IL17 levels indicating its potential pathogenic importance in RA and may represent an attractive target for future therapeutic interventions.     

The relationship between disease activity and depression in Egyptian patients with rheumatoid arthritis

Aim of the workTo estimate the prevalence of depression and its relationship with disease activity parameters in Egyptian patients with RA.Patients and methodsA cross sectional study was conducted on 170 patients with RA. The following values were assessed for each patient: erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), swollen and tender joint counts (SJC and TJC), disease activity score 28 (DAS28), health assessment questionnaire score (HAQ), visual analogue scale (VAS) of pain and hospital anxiety and depression scale-depression subscale (HADS-D).ResultsThe prevalence of depression was 15.29% (26 RA patients). In the depressed RA patients, positive significant correlations were found between HADS-D score and age, disease duration, HAQ score, VAS, DAS28 score and CRP. However, no significant correlation was found between HADS-D score and ESR, number of swollen and tender joints. No significant difference (P > 0.05) was found between depressed male and female patients with RA.ConclusionPatients with RA and co-morbid depression have worse health outcomes. RA cases should be monitored for accompanying depression during follow-up. The identification and treatment of depression in RA paramount to the overall management of RA.     

Leptin serum levels in rheumatoid arthritis patients: relation to disease duration and activity

Leptin is a peptide hormone with the tertiary structure of a cytokine, which not only regulates body weight by inhibiting food intake, but also modulates inflammatory and immune responses. The aim of the study was to investigate if there are connections between leptin concentrations and parameters of nutritional status and disease activity in a group of rheumatoid arthritis (RA) patients. The study group consisted of 37 patients. The mean leptin serum concentration was significantly higher in women than in men. The leptin concentrations correlated positively with BMI only in women with RA. The leptin concentrations were significantly higher in patients with erosive RA. Assessing the group of patients with long-standing RA (duration > 10 years), we found that leptin levels were significantly higher in patients with higher disease activity than in those with DAS28 ≤ 5,1; there was also a positive correlation between serum leptin concentration and the value of DAS28, ESR and the number of tender joints. The results suggest that some important dependence exists between the risk of aggressive course of RA and increased leptin levels.     

Evaluation of serum calprotectin level as a biomarker of disease activity in rheumatoid arthritis and osteoarthritis patients

Aim of the workTo measure the levels of serum calprotectin (CLP) in rheumatoid arthritis (RA) and osteoarthritis (OA) patients and to assess its association with disease activity, severity and functional status.Patients and methodsA total of 30 RA and30 OA patients and 30 controlswere included. Rheumatoid factor (RF), anti-cyclic citrullinated peptide (anti-CCP), Disease activity score (DAS28), health assessment questionnaire (HAQ) and RA medical records-based index of severity (RARBIS) were assessed in RA patients. Western Ontario and McMaster Osteoarthritis index (WOMAC) and Kellgren-Lawrence (KL) grading scale were assessed in OA patients and serum CLP levels were measured.ResultsThe mean age of RA and OA patients was 48.6 ± 8.6 and 50.8 ± 9.3 years respectively andthe majority of studied groups were females. CLP was significantly higher in RA patients in comparison to OA patients and healthy control (2.70 ± 2.08 vs. 1.18 ± 0.35 vs 1.11 ± 0.24 μg/ml); p < 0.0001). Serum CLP correlated with swollen joint count (SJC) (r = 0.7, p < 0.0001), tender joint count (TJC) (r = 0.73, p < 0.0001), patient global assessment (PGA) (r = 0.51, p = 0.004), Physician global assessment (PhGA) (r = 0.58, p = 0.001), HAQ (r = 0.6,p < 0.0001), erythrocyte sedimentation rate (ESR) (r = 0.5, p = 0.005), DAS28 (r = 0.69, p < 0.0001), RARBIS (r = 0.66, p < 0.0001). At a cut-off value of 2.5 µg/ml CLP can significantly differentiate active RA patients from those in remission (AUC 0.896; p < 0.0001) at a sensitivity of 83.3%, specificity of 88.9%, and accuracy of 86.7%.CLP was significant predictor for RA activity.ConclusionThe serum CLP levels were significantly high in RA patients compared to OA patients and controls and these high levels were associated with disease activity, severity, and functional status.     

Analysis of cytokine production patterns of peripheral blood mononuclear cells from a rheumatoid arthritis patient successfully treated with rituximab

Abstract

We had a rheumatoid arthritis (RA) patient resistant to multiple drugs and who developed panniculitis due to etanercept treatment, then responded fairly well to rituximab. Intracellular staining of cytokines in the peripheral blood mononuclear cells before and after rituximab administration revealed that the cytokine production, representative of T-helper (Th)1-, Th2-, and Th17-type responses, decreased abruptly after the treatment. Interestingly, this timing coincided with that of the manifestation of the beneficial effect. This relationship may provide useful insight into the mechanism of action of the drug and hence about the pathogenesis of RA.     

Serum matrix metalloproteinase-3 in rheumatoid arthritis patients: Correlation with disease activity and joint destruction

Aim of the workThis study was designed to measure the serum level of matrix metalloproteinase-3 (MMP-3) in rheumatoid arthritis (RA) patients and its correlation with functional status, disease activity and joint damage.Patients and methodsThe study included 50 RA patients satisfying 2010 ACR/EULAR classification criteria recruited from Bani-Suef University Hospital and 20 controls. Functional disability was assessed according to Modified Health Assessment Questionnaire (MHAQ). Disease activity score in 28-joints (DAS28) and visual analogue scale (VAS) of pain were evaluated. Radiological joint damage was assessed by Van der Heijde-modified Sharp Score (vdHSS). Serum levels of MMP-3 were measured for all subjects.ResultsRA patients (44 females and 6 males) had a mean age of 46.36 ± 13.63 years and disease duration of 5.6 ± 4.75 years. Serum MMP-3 levels were higher in patients than in controls (46.78 ± 46.99 versus 1.98 ± 1.71 ng/ml respectively, p = 0.0001) and significantly correlated with erythrocyte sedimentation rate (p = 0.001) and were significantly higher in patients with positive C-reactive protein, rheumatoid factor and anti-cyclic citrullinated peptide (p = 0.0001, p = 0.009, p = 0.042, respectively). MMP-3 significantly correlated with DAS28 (p = 0.0001) and vdHSS (r = 0.78, p = 0.0001) and a significant difference was shown in those with erosions compared to those without (p = 0.001). Serum MMP-3 levels significantly correlated negatively with cumulative steroid dose (r = −0.2, p = 0.03) and were significantly higher in patients who never received disease-modifying antirheumatic drugs (p = 0.001). There were no significant relations of MMP-3 with age, MHAQ, VAS for pain.ConclusionThese results indicate that serum MMP-3 is a measurable, useful specific marker of disease activity and joint damage in RA patients.     

类风湿关节炎患者外周血Th17细胞的检测及意义

目的 探讨Th17细胞在类风湿关节炎(RA)患者外周血中的水平及意义.方法 分离RA患者和健康者外周血单个核细胞(PBMC),免疫磁珠阴选CD4~+T细胞,用或不用非特异性刺激剂(A-CD3、A-CD28),然后加佛波酯/离子霉素(PMA/Ion),经固定,透膜处理进行细胞内染色,流式细胞术(FCM)检测CD4~+T细胞内白细胞介素(IL)-17~+/干扰素(IFN)-γ~+、IL-17~+/IL-6~+水平.分组:①健康对照组;②RA病情稳定组;③RA病情活动组.结果 免疫磁珠阴选CD4~+T细胞纯度>90%.RA病情活动组IL-17表达水平(1.54±0.41)显著高于RA病情稳定组(0.70±0.21,P<0.01),二者又显著高于健康对照组(0.42±0.12,P<0.01).用A-CD3、A-CD28、IL-23刺激后,CD4~+T细胞IL-17胞内表达水平较无刺激组显著增加(P<0.05).CD4~+T细胞IFN-γ表达水平呈现与IL-17表达相似的特点,而RA患者与健康对照组IL-6表达水平差异无统计学意义.RA患者IL-17胞内表达水平与IFN-γ、IL-6无显著相关.结论 RA患者外周血CD4~+T细胞中存在异常增高的Th17细胞,且其水平与病情活动相关,有望作为判断RA患者病情活动的指标之一.