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A proteomic study of peripheral blood mononuclear cells in systemic lupus erythematosus

Chengxiao HU ; Yong DAI ; Jianjun LIU ; Jianfan HE ; Tianyu LV

Chinese Journal of Rheumatology.2009;13(11):779-782. doi:10.3760/cma.j.issn.1007-7480.2009.11.014

Objective To analyze the changes in the protein expression profile of peripheral blood mononuclear cells in systemic lupus erythematosus patients. Methods Peripheral blood was obtained from SLE patients and healthy controls, then mononuclear cells were isolated and the total protein was extracted by one-step method. Two-dimensional gel electrophoresis was performed and then stained with silver. Protein maps were analyzed and differentially expressed protein spots were detected using ImageMaster 2D Platinum 5.0 software. Results Match rates of (71±4)% and (72±4)% was obtained from gels from controls and pati-ents respectively. 791±17 spots were detected from control gels and 781±17 from patient gels. Eleven protein spots were up-regulated and 9 were down-regulated in SLE patients. Five proteins were identified by MS analysis, some of which had previously been shown to play a potential role in the pathogenesis of SLE. Conclusion There are significant changes in the protein expression of peripheral blood mononuclear cells in systemic lupus erythematosus patients. This study could be used as a preliminary work for better understanding of the pathogenesis and immune regulation pathways of SLE from an integrated lymphocyte protein profile perspective.

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Expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis

Jinbing RUI ; Jing LI ; Haiyan YOU ; Ling WU ; Yingying QIU ; Yanping LUO ; Xiaoli SONG ; Zhijun JIAO

Chinese Journal of Rheumatology.2009;13(11):732-735. doi:10.3760/cma.j.issn.1007-7480.2009.11.002

Objective To study the expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis (RA). Methods Intracelluar flow cytomete detection of IL-17/IFN-γ and IL-17/IL-6 was established using anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. There were three groups in the present study: ①healthy controls group; ② RA stable group; ③RA active group. Results The isolation of untouched human CD4~+T cells from PBMC was effective and its purity was over 90%. The percentage of intracelluar IL-17 in CD4~+ T cells from RA patients was increased significantly. Such percentage in active group (1.54±0.41) was higher than that of stable group (0.70±0.21, P<0.01) and both of them were higher than those of healthy controls (0.42±0.12, P<0.01). Under anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of intracelluar IL-17 was also increased significantly(P<0.01). The porcentage of intracellular IFN-γ was similar to that of IL-17, while that of IL-6 was not significantly different. There is an correlation between IL-17 and IFN-γ or IL-6. Conclusion There is an abnormal expression of IL-17 and IFN-γ in human CD4~+T cells in RA patients, which is related to disease activity . Th17 cells may be used as a new marker for the assessment of RA activity.

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Significance and alteration of T help 17 cells in immune globulin-resistant gawasaki disease

Shilei JIA

Chinese Journal of Rheumatology.2009;13(11):736-740. doi:10.3760/cma.j.issn.1007-7480.2009.11.003

Objective To investigate the alteration and significance of T help 17 cells (Th17), in patients with intravenous immune globulin-resistant Kawasaki disease (KD). Methods Forty-five children with KD (thirty-five as sensitive group and ten as the resistant group) and thirty age-matched healthy children were studied. Real-time PCR was used to evaluate the mRNA levels of IL-17A/F, ROR-γt. The proportion of Th17 in peripheral blood was detected by flow cytometric analysis. Th17-related plasma cytokine (including IL-17A and IL-6) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Results ① Compared with healthy eontrols, transcription levels of IL-17A/F were significantly upregulated during acute phase of KD (P<0.01). ② Compared with sensitive group, the transcription levels of IL-17A/F, ROR-γt were significantly up-regulated during acute phase in immune globulin-resistant KD (P< 0.01), and down-regulated after treatment with intravenous immunoglobulin (IVIG) therapy in sensitive group (P<0.01), but there was no significant difference in the levels of expression in resistant group (P>0.05). There was significant difference in serum IL-17A,IL-6 before or after IVIG between resistant and nonresistant group (P<0.01). ③ Compared with the normal controls, serum IL-17A, IL-6 were markedly elevated in patients with KD. There was significant difference in serum IL-17A and IL-6 concentrations between resistant and sensitive group before IVIG treatment (P<0.01). The resistant group showed ignificantly high IL-17A, IL-6 after IVIG. There was no significant difference in the severity of decrease before and after IVIG in both groups, but it was found that serum IL-17A and IL-6 had a tendency to decrease after treatment with IVIG. ④ Methyiprednisolone might down-regulate inflammatory response through inhibiting the secretion of inflammatory IL-6 and IL-17A cytokines in acute stage of immunoglobulin-resistant KD. Treatment of immunoglobulin-resistant Kawasaki disease with IVMP resulted in faster resolution of fever, more rapid improvement in markers of inflammation. Conclusion Aberrant activation of Th17 cells may be one of the factors causing disturbed immunological function in immunoglobulin-resistant KD.

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Thalidomide successfully maintains remission in ankylosing spondylitis after discontinuing etanercept treatment

Xiaohu DENG ; Feng HUANG ; Jianglin ZHANG ; Jie ZHANG ; Yamei ZHANG

Chinese Journal of Rheumatology.2009;13(11):765-768. doi:10.3760/cma.j.issn.1007-7480.2009.11.010

Objective To determine whether thalidomide can maintain remission after discontinuing the treatment of etanercept in ankylosing spondylitis (AS). Methods One hundred and five patients with active AS treated with etanercept 50 rag/week for 12 weeks and attained an ASAS20 response at week 12. They were randomly assigned to receive thalidomide 150mg/night, sulfasalazine (SASP) 2.0 g/d, or non-steroidal anti-inflammatory durgs (NSAIDs) only. All patients were followed monthly for BASDAI, BASFI, PGA and spinal pain VAS. A Kaplan-Meier survival analysis was used to calculate the probability of a relapse. Results One hundred patients completed the follow-up. Thirty patients were treated with thalidomide, 33 patients with SASP and 37 patients with NSAIDs only. The mean follow-up time was 5.1 months and the longest time was 12 months. At the end of the follow-up, the percentage of patients who maintained remission in the thalidomide group was 40%, much higher than SASP group (15%) and NSAIDs group 11% (P=0.0265 and 0.0053 respectively). No difference was found between the remission rate of SASP and NSAIDs only group (P=0.5881). Conclusion Thalidomide can successfully maintain remission of AS after discontinue etanercept treatment.

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The therapeutic effect of etanercept on intractable ankylosing apondylitis related hip joint diseases

Li MA ; Dongxue WANG ; Donghai WU

Chinese Journal of Rheumatology.2009;13(11):754-756. doi:10.3760/cma.j.issn.1007-7480.2009.11.007

Objective To observe the therapeutic effect of tumor necrosis factor inhibitor (Etanercept) on intractable ankylosing spondylitis (AS) related hip joint lesion. Methods Thirty-five patients with AS with unilateral or bilateral hip joints pain and limitation of joint motion were included into this study. The patients' conditions were not controlled under standard treatment by non-steroidal anti-inflamma-tory drug and antirheumatic medications. The clinical trial was designed as a prospectiveopen study, 35 pati-ents received Etanercept 50 mg once a week for 12 weeks, combined with methotrexate (MTX) 10 mg once a week. Parameters including Harris hip score, Bath ankylosing spondylitis radiologic index-hip (BASRI-hip), Bath ankyiosing spondylitis disc.use activity index (BASDAI), Bath ankylosing spondylitis functional index (BASFI), Erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) were evaluated and side effects were observed before and after the treatment. Results Fifty-five hip joints were involved in 35 patients, in which unilateral hip involvement in 15 patients and bilateral in 20 patients. Harris score of the hips iner-eased significantly from 51±4 before treatment to 86±5 (P=0.000) after treatment ; Before and after treatment, BASDAI changed from 6.4±1.2 to 4.4±0.8 (P=0.000), BASFI was changed from 6.3±1.1 to 3.4±0.8 (P=0.000), before and after treatment ESR was changed from (68±28) mm/l h to (25±6) mm/l h (P=0.001), CRP changed from (59.1±22.3) mg/l, to (6.9±1.1) mg/L (P=0.000) before and after treatment respectively, but BASRI-hip was not changed obviously before and after treatment. No tuberculosis and serious side effects was observed during the treatment and follow-up period. Conclusion Etanercept, when combined with methotrexate, could be used to treatintractahle AS-related hip joint lesions. This regimen could improve the hip joint function and control the disease activity without serious side effects.

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The clinical predictive value analysis of multi-autoantibodies detection in the transformation from undifferentiated arthritis to rheumatoid arthritis

Zhu CHEN ; Yufeng YANG ; Caihong WANG ; Jieting JIA ; Lin ZHANG ; Zhen ZHANG ; Zhiyong JIN ; Guangwen AN ; Xiaofeng LI

Chinese Journal of Rheumatology.2009;13(11):749-753. doi:10.3760/cma.j.issn.1007-7480.2009.11.006

Objective To investigate the clinical predictive value of rheumatoid factor (RF), anti-keratin antibodies (AKA), anti-cyclic citrullinated peptide (anti-CCP) antibody and anti-perinuclear factor (APF) in the transition from undifferentiated arthritis (UA) to rheumatoid arthritis (RA), and to analyze the clinical relevant factors. Methods 271 patients with UA who were followed up for a year were enrolled into the investigation. RF was measured by the rate scatting immunity method. APF and AKA were detected by immuniofluorescence method(IFA). Anti-CCP was measured by ELISA. Erythrocyte sedimentation rate (ESR) was analyzed by Wilcoxon method. The duration of morning stiffness, the numbers of swelling and tender joints, tender joints involved and DAS28 score were recorded and analyzed. Results 99% patients who had four-antibody-positive finally convetted from UA to RA. The conversation rate for those who had two or more than two antibody-positive was 83.0% and 65.9% respectively. The sensitivity and specificity of RF and anti-CCP antibody-positive in those who converted from UA to RA was 77.8% and 80.5% respectively. The percentage of polyarticular swelling in antibody-negative, one-antibody-positive, two-antibody-positive, three-antibody- positive and four-antibody-positive was 48%, 57%, 59%, 70% and 70% respectively. Meanwhile, the percentage of multi-small-joint involvement was 71%, 71%, 72%, 76% and 83% respectively. The proportion of elbow involvement in antibody-negative patients was 72%, which was the highest among all joint area involvement. The conversion differences of the group with more than 3 swelling joints or more than 3 small joints involvement ranked the first and second in frequency. Conclusion The combined detection of these autoantibodies could increase the specificity of early diagnosis of RA. The more positive antibodies present, the more likely the concersion form UA to RA. The sensitivity and specificity of RF and anti-CCP-positive is high, so the latter is expected to become one of the diagnostic criteria. The polyarticular swelling and multi-small-joint involvement are valuable in predicting the transformation from UA to RA.

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The expression of adiponectin and its receptors in the inflammatory joint of rheumatoid arthritis patients

Fang WANG ; Wenfeng TAN ; Miaojia ZHANG ; Dunming GUO ; Xiaohua LIU ; Youxuan SHEN ; Yao KE ; Shaoheng HE

Chinese Journal of Rheumatology.2009;13(11):745-748. doi:10.3760/cma.j.issn.1007-7480.2009.11.005

Objective To study the expression of Adiponectin (AD) and its receptors Adiponectin receptor 1 (Adipo R1) and Adipo R2 in the synovial fluids and the synovium of rheumatoid arthritis (RA). Methods ELISA was used to determine the levels of AD in 23 RA and 23 osteoarthritis (OA) patients. Real-time PCR and Western blot techniques were employed to study the expression of AD, AdipoR1 and AdipoR2 in the synovium of 10 RA and OA patients. Results It was observed that approximately twice more adiponeetin in the synovial fluids of patients with RA than with OA. Adiponectin and AdipoR1, but not AdipoR2 mRNA, were significantly expressed in synovium of RA patients in comparison with OA. Adiponectin and AdipoR1 protein were wuch more expressed in synovium from RA than those from OA. Conclusion High expression of Adiponectin and AdipoRl is likely to contribute to the formation of inflammatory joints in RA.

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The significance of serum SC5b-9, anti-Clq antibody, C3 and C4 levels in assessing lupus disease activity

Zhijing QIAN ; Rongfu ZHANG ; Li MA ; Donghai WU

Chinese Journal of Rheumatology.2009;13(7):459-462. doi:10.3760/cma.j.issn.1007-7480.2009.07.008

Objective To explore the value of serum SC5b-9, anti-C1q antibody, C3 and C4 levels in the assessment of lupus activity. Methods The enzyme linked immunosorbent assay (ELISA) was used to measure SC5b-9 and anti-C1q antibody, rate nepheiometry was used to detect the serum level of C3 and C4 in sera of 62 SLE patients, 35 patients with other rheumatic diseases (including rheumatoid arthritis, ankylosing spondylitis, primary Sjogren' s syndrome, mixed connective tissue disease, dermatomyositis, polymyositis, systemic sclerosis and vasculitis) and 35 healthy controls. And the correlation between above-mentioned parameters and lupus clinical manifestations, disease activity and histological type of lupus nephritis were analyzed. Results In SLE patients, the levels of SC5b-9 and anti-C1q antibody were significantly higher than those in patients with other rheumatic diseases and healthy controls (P<0.05). The titers of SC5b-9 and anti-C1q antibody negatively correlated with C3 and C4 (P<0.05), and positively correlated with SLEDAI (P<0.05). The sensitivity and specificity of the combination of these three measurements for SLE was 95.37% and 98.46 respectively. SC5b-9 and anti-C1q antibody were associated with the presence of proliferative glomerulonephritis (P <0.05). Conclusion Taking the evaluation of all these three measurements simultaneously is valuable for the diagnosis of lupus flare. SC5b-9 and anti-C1q antibody may play major roles in the immunopathogenesis of lupus nephritis.

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Quality of life assessment and related factors of patients with active rheumatoid arthritis

Hongqing NIU ; Xiaofeng LI ; Haiyuan DONG ; Jinli RU ; Liyun ZHANG ; Jinfang GAO

Chinese Journal of Rheumatology.2009;13(7):447-450. doi:10.3760/cma.j.issn.1007-7480.2009.07.005

Objective To analyze the quality of life of patients with active rheumatoid arthritis (RA) and its relationship with other clinical and functional parameters used for the evaluation of disease activity. Methods The quality of life was assessed in 127 patients with active RA using SF-36 and was compared with non-active RA and the general population. The correlation between the quality of life and the clinical measures of disease, including morning stiffness, pain, fatigue, patient's global assessment (PGA) physician's global assessment , SJC/SJ1, TJC/TJI, DAS28, HAQ were measured. Results The patients with active RA reported significantly decreased scores in all dimensions of SF-36. Fatigue, PGA, physician's global assess-ment, DAS28 and HAQ significantly correlated with the scores in all dimensions of SF-36. Pain was correlated with the scores in all dimensions of SF-36 except RE. TJI was correlated with six dimensions. TJC was correlated with five dimensions. ESR was correlated with three dimensions. Conclusion The quality of life in patients with active RA is significantly declined compared with non-active RA and the general population. Disease activity and functional status of patients with active RA is closely correlated with the quality of life.

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Overwhelming Th17 cells compared to insufficient CD4+ regulatory T cells in rheumatoid arthritis

Tian WANG ; Yi ZHAO ; Xu LIU ; Xia LIU ; Ru LI ; Limin REN ; Hua YE ; Zhanguo LI

Chinese Journal of Rheumatology.2009;13(7):439-442. doi:10.3760/cma.j.issn.1007-7480.2009.07.003

Objective To identify interleukin (IL)-17-producing T cells and Foxp3 positive CD4 positive T ceils from patients with rheumatoid arthritis (RA), and investigate their cytokine levels as well as their correlations. Methods Flow cytometry was used to analyze the subsets of T cells in peripheral blood from 39 RA patients and 40 healthy donors. IL-17, IL-6, IL-23 and transforming growth factor (TGF)-beta levels in sera of these subjects were tested by ELISA. Results IL-17+CD4+ T cells increased significantly and IL-17, IL-6 and IL-23 elevated markedly in peripheral blood of RA patients compared to healthy donors (P< 0.01 ). However,Foxp3+CD4+T cells decreased evidently (P<0.01) and TGF-beta did not increase in RA pati-ents when compared to healthy donors (P>0.05). Conclusion This study suggests that Th17 cells predomin-ance coupled with relatively insufficient CD4+ regulatory T cells in RA is mainly caused by alterations of rela-ted cytokines.

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