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Introducción: La artritis reumatoidea se caracteriza por inflamación de la membrana sinovial debido al infiltrado de células inmunitarias que secretan citocinas relacionadas a perfil Th17 como IL-22 e IL-6. La dinámica de estas citocinas durante el tratamiento permanece incomprendida. El objetivo fue evaluar los niveles séricos y en líquido sinovial (LS) de IL-22 e IL-6, correlacionarlos con diferentes parámetros bioquímicos y clínicos y medir sus cambios post-tratamiento. Material y métodos: Se estudiaron 77 pacientes con AR y 30 controles. A 30 pacientes se los evaluó nuevamente luego de 3 meses de tratamiento y a 12 se les extrajo LS. Se midió VSG, PCR, FR, anti-CCPhs, IL-22 e IL-6. Se evaluó la actividad con DAS28 y respuesta al tratamiento con criterios EULAR. Resultados: IL-22 e IL-6 fueron similares entre pacientes y controles. Sus niveles disminuyeron luego del tratamiento, principalmente en pacientes respondedores. IL-22 fue menor e IL-6 mayor en LS que en sangre. IL-6 correlacionó positivamente con PCR y anti-CCPhs. Los niveles de VSG, PCR y DAS28 fueron mayores en pacientes con valores dosables de IL-6 que en no dosables. Conclusión: En pacientes con valores basales dosables de IL-22 e IL-6, los niveles de estas citocinas podrían utilizarse como marcador adicional de respuesta al tratamiento.
Introduction: Rheumatoid arthritis is characterized by synovium inflammation due to the infiltration of immune cells that secrete Th17 cytokines like IL-22 and IL-6. The dynamics of these cytokines during the treatment remain unknown. The aim of this study was to evaluate the levels of IL-22 and IL-6 serum and synovial fluid (SF) in correlation with different biochemical and clinical parameters and treatment-associated changes. Material and methods: Seventy-seven RA patients and 30 controls were recruited. Thirty patients were evaluated after 3 months of treatment and SF was collected of 12 patients. ESR, CRP, RF, anti-CCP hs, IL-22 e IL-6 were measured. DAS28 was used to assess disease activity and response to treatment followed EULAR criteria. Results: There were not differences in serum IL-22 and IL-6 levels between patients and controls. Cytokine levels decreased after treatment, mainly in responder patients. IL-22 was decreased and IL-6 was increased in SF compared to serum. IL-6 correlated positively with CRP and anti-CCPhs. ESR, CRP and DAS28 were increased in patients with detectable IL-6 compared to those with undetectable IL-6. Conclusion: In patients with detectable serum IL-22 and IL-6 levels before treatment initiation, follow-up of cytokine levels could be an useful additional tool to evaluate treatment response.
Subject(s)
Arthritis, Rheumatoid , Therapeutics , Interleukins , Interleukin-6 , InflammationABSTRACT
Rheumatoid arthritis (RA), psoriatic arthritis (PsA), and ankylosing spondylitis (AS) are inflammatory diseases with different bone remodeling patterns. Fibroblast-like synoviocytes (FLS) are cells involved in the transition from an acute and reparable phase to a chronic and persistent stage in these diseases. The distinction of joint phenotypes involves inflammatory cytokines such as tumor necrosis factor alpha (TNF-α), interleukin (IL)-17, and IL-22 directly or through key signaling pathways such as Wnt. To evaluate the role of FLS as the source of Wnt antagonists (sFRP3/FRZB and Dkk1) in the synovia, levels of TNF- α, IL-17, IL-22, Dkk1, and sFRP3 were measured by ELISA directly in the synovial fluid of patients with RA, PsA, or AS. Dkk1 and sFRP3 were also measured in the FLS culture supernatants after different inflammatory stimulus. sFRP3 and Dkk1 are constitutively expressed by FLS. IL-22 and sFRP3 were positively correlated (r=0.76; P<0.01) in synovial fluid. The stimulation of FLS with IL-22, but not TNF-alpha and IL-17, increased the production of sFRP3. No stimulus altered the basal expression of Dkk1. These results showed, for the first time, the ability of IL-22 to increase the expression of sFRP3/FRZB by human FLS in both in vitro and ex vivo models. This finding linked IL-22 to local inhibition of Wnt signaling and possibly to blockade of osteogenesis. Furthermore, FLS presented as a source of this inhibitor in synovial fluid, assigning to this cell a bone injury mechanism.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Interleukins/metabolism , Synoviocytes , Synovial Membrane , Cells, Cultured , Tumor Necrosis Factor-alpha , FibroblastsABSTRACT
Objective To analyze the frequency of interleukin (IL)-22+CD161+CD4+ T cells in the peripheral blood mononuclear cells (PBMCs) in rheumatoid arthritis (RA) patients compared with healthy control subjects and investigate the relationship of IL-22+CD4+CD161+ T lymphocyte frequency changes with RA disease activity.In addition to explore the pathogenesis of RA,and to look for new treatment targets for RA.Methods Twenty-one RA cases were included in the Department of Rheumatology of Tangshan Gongren Hospital from 2017 to 2018.Fourteen patients were female and 7 were male with the age ranged from 36 to 74 years old.The average age of this group of patients was (55±10) years,the average disease course was (60±50) months.All patients fulfilled the classification criteria of American College of Rheumatology [American College of Rheumatology (ACR)].Twenty-one subjects were enrolled as the control group,all of them came to Tangshan Gongren Hospital for regular health check-up.Fifteen subjects in the control group were female and 6 were male.Their age ranged between 40-78 years old with the average age of (55±9) years.IL-22+CD4+CD161+ T cells in PBMCs were detected by flow cytometry.The frequency variation of different CD4+CD161 + T was compared between case and control groups.The correlation was studied between the frequency and RA disease activity score (DAS28),tender joints number,swollen joints number,red blood cell sedimentation rate,high sensitive C reactive protein and white blood cell counts,red blood cell counts,platelet counts,IgG,IgA,IgM,complement C3 level,complement C4 level.T-test or Mann-Whitney U test were used for single-factor analysis,Pearson's test was used for correlation analysis.Results The percentage of RA group secreted CD4+ T cells (0.33± 0.20)％ of INF-γand IL-22,CD4+ T cells (0.51±0.29)％ of IL-22,and CD4+CD161+ T cells of IL-22 simultaneously.The number (0.55 ±0.28)％ was.significantly higher than that of the healtby control group [(0.22±0.14)％,(0.25±0.18)％,(0.36±0.24)％],and the differences were statistically significant [P=0.002,P=-0.0.45,P=0.026].Conclusion The percentage of IL-22+CD4+CD161+ T lymphocytes in the peripheral blood monocytes in RA patients is significantly higher than that in the healthy controls.The results of this study suggest that IL-22+CD4+CD161+ T lymphocytes in RA patients maybe related to RA disease activity and joint lesions.
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Objective To study the significance of serum interleukin(IL)-18,IL-22 and IL-23 levels in rheumatoid arthritis.Methods 40 patients of rheumatoid arthritis w ho received therapy from December 2014 to December 2016 in our hospital were selected,and 40 cases of healthy people in our hospital for the same pe-riod were selected as a control group,the expressions of serum IL-18,IL-22 and IL-23 were compared between the two groups;The relationship between the serum IL-18,IL-22,IL-23 and the severity of rheumatoid arthri-tis,before and after treatment and clinical indicators were analyzed.Results The serum IL-18[(741.82 ± 45.60)pg/mL],IL-22[(62.34 ± 3.72)pg/mL]and IL-23[(141.73 ± 18.31)pg/mL]in rheumatoid arthritis group were higher than that of control group[(231.28 ± 26.71),(37.26 ± 3.91),(48.28 ± 5.70)pg/mL],and the difference has statistical significance(P<0.05);the serum IL-18,IL-22 and IL-23 in highly active group rheumatoid arthritis patients were significantly higher than that of moderate active group and stable/mild ac-tive group,serum IL-18,IL-22 and IL-23 in the moderate active group were significantly higher than that of stable/mild active group,and the difference has statistical significance(P<0.05);after treatment,the serum levels of IL-18,IL-22 and IL-23 in patients with rheumatoid arthritis were significantly lower than those before treatment,and the difference has statistical significance(P< 0.05);the results of correlation analysis show that there was a positive correlation between the serum IL-18 and joint tenderness index,joint swelling index, C reactive protein and bone destruction score(P<0.05),and there was a positive correlation between the ser-um IL-22,IL-23 and VAS score,the time of morning stiffness,joint pain index,swelling index,platelet count,C reaction protein and done destruction score(P<0.05).Conclusion The serum IL-18,IL-22,and IL-23 are closely related to the severity of rheumatoid arthritis,which can increase with the aggravation of disease,and it also helps to assess the effectiveness of disease treatment,application value is high.
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Objective:To explore the expression of IL-22 in the patients with rheumatoid arthritis,and to define the clinical sig-nificance of IL-22 cells for RA. Methods: A total of 50 RA patients, 15 OA patients and 15 healthy controls were enrolled. The proportion of Th22 cells in peripheral blood and synovial fluid( SF) of RA patients was detected by flow cytometry;the levels of IL-22 in serum and synovial fluid of RA patients were detected by ELISA. The clinical parameters of disease activity were assessed including ESR,RF,CRP,DAS28,anti CCP and the degree of bone erosions determined by X-rays. Pearson correlation analysis,t test and Kruskal-Wallis H test were used for statistical analysis. Results:The proportion of Th22 cells in RA patients was higher than that of OA patients (t=2. 290 ,P=0. 021) and healthy controls(t=2. 524,P=0. 015). IL-22 levels in RA patients were higher than that of OA patients (t=2. 560,P=0. 014) and healthy controls(t=2. 768,P=0. 009). IL-22 in the RF positive group were higher than that of RF negative group(t=2. 322,P=0. 035). IL-22 in the anti-CCP antibody positive group were higher than that of anti-CCP antibody negative group (t=2. 504,P=0. 015). The levels of IL-22 were correlated positively with ESR,RF,DAS28(r=0. 312,0. 314,0. 332,P χ20.05(3),P<0. 05). Serum IL-22 levels in the RA patients with joint effusion were higher than that of without(t=2. 587,P=0. 012). The levels of IL-22 in SF were higher than that in serum(t=2. 668,P=0. 011),and had no correlation with the proportion of Th22 cells in SF. Conclusion: The expression of IL-22 in serum and joint effusion of RA patients increase. The level of IL-22 may be useful marker for assessment of disease activity and finding of bone erosions. Therapeutic targeting of IL-22 may be valuable for the intervention of RA.
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AIM:To determine the effects and mechanisms of interleukin-22 (IL-22) on the fibroblast-like sy-noviocytes ( FLSs) from rheumatoid arthritis ( RA) patients.METHODS:RA-FLSs were cultured by tissue culture meth-od.RA-FLSs were incubated with different concentrations of IL-22 (0,1,10,100μg/L) for 24 h, 48 h and 72 h.The cell viability was examined by CCK-8 assay.IL-22 at concentration of 10 μg/L was used to stimulate RA-FLSs for 24 h, and the change of cell cycle distribution was identified by flow cytometry.The effects of IL-22 at concentrations of 0, 1, 10, 100μg/L and/or STA-21 (a STAT3 inhibitor at concentrations of 0, 25, 50μmol/L) on the protein levels of Bcl-2 and p-STAT3 in the RA-FLSs were determined by Western blot.RESULTS:Compared with control group, stimulation of rhIL-22 at different concentrations for 24 h, 48 h and 72 h, the cells viabilityof RA-FLSs were obviously increased ( P<0.05 ) . After co-cultured with 10 μg/L rhIL-22 for 24 h, the percentages of RA-FLSs were obviously increased in the G2/M+S phase and decreased in the G0/G1 phase.At the same time, rhIL-22 increased, but STA-21 decreased the protein levels of Bcl-2 but p-STAT3 in the RA-FLSs obviously (P<0.05).Treatment with STAT3 inhibitor STA-21 reversed the effect of IL-22-induced Bcl-2 upregulation in the RA-FLSs ( P<0.01 ) .CONCLUSION: STAT3 is critical in the process of IL-22-induced Bcl-2 upregulation in RA-FLSs, indicating that IL-22 may play a role in the apoptosis of RA-FLSs.
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Objective To study the relationship between the T helper cells (Th22)/interleukin (IL)-22 and rheumatoid arthritis (RA) with interstitial lung disease (ILD), and to define the clinical significance of Th22 cells for RA. Methods The quantity of Th22 cells in the peripheral blood from 40 patients with RA (20 RA with ILD, 20 RA without ILD) were examined by flow cytometry, the level of IL-22 in the sera was detected by enzyme-linked immunosorbent assay (ELISA). Comparisons between groups were analyzed by t-test, rank sum test, and the correlation of parameters were tested by linear correlation analysis. Results The quantities of CD4+IL-22+ cells (Th22) in RA patients [(0.15 ±0.07)%] were significantly higher than normal controls [(0.09 ±0.05)%] (t=4.097, P<0.01), and IL-22 levels in RA patients [(83 ±7) ng/L] were significantly higher than normal controls [(61±5) ng/L] (t=13.057, P<0.01). The quantities of Th22 cells in RA-ILD patients [(0.18±0.07)%] were significantly higher than RA-NILD patients [(0.13±0.05)%] (t=2.919, P=0.008), and IL-22 levels in RA-ILD patients [(87±6) ng/L] were significantly higher than RA-NILD patients [(80±6)ng/L] (t=3.624, P=0.001). The quantities of Th22 cells were positively correlated with erythrocyte sedimentation rate (ESR), rheumatoid factor(RF) and 1.4 disease activity score (DAS)28 (r=0.336, 0.377, 0.577, P<0.05),and the level of IL-22 were also positively correlated with ESR and DAS28 (r=0.406, 0.576, P<0.05). Conclusion The quantities of Th22 cells and IL-22 level are increased in RA patients, especially in RA-ILD patients. The quantities of Th22 cells and IL-22 level are positively correlated with ESR and DAS28. It may play a certain role in RA especially in RA with ILD.
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Objective To investigate the effects of interleukine-22 ( IL-22 ) on the expression of interleukin-6 (IL-6) by rheumatoid arthritis synovial fibroblasts (RASF), and to analyze their association with IL-17+CD4+T (Th17) cells differentiation.Methods RASF were isolated from six patients with rheu-matoid arthritis ( RA) and cultured in vitro.The expression of IL-6 at mRNA and protein levels by RASF were detected by qRT-PCR analysis and ELISA after treatment with different concentrations of IL -22 for dif-ferent periods of time.Anti-IL-22R1 blocking antibody and inhibitor assay were used to analyze the specific receptor and its downstream signaling pathways associated with IL-6 production.IL-22 pre-treated RASF and CD4+T cells were co-cultured for 3 days in the presence or absence of anti-IL-22R1 or anti-IL-6 to measure the percentage of Th 17 cells by flow cytometry .Results The expression of IL-6 by RASF was increased up-on IL-22 stimulation in a dose and time dependent manner (P<0.05), and that was closely related to IL-22R1 and its downstream signaling pathways of p38 and JAK2 (P<0.05).Co-culturing CD4+T cells with RASF and Transwell system indicated that the percentage of Th 17 cells was increased in IL-22 pre-treated group as compared with that in IL-22 untreated group , but it could be down-regulated by either blocking IL-22R1 or IL-6.Conclusion IL-22 promoted the expression of IL-6 by RASF and further enhanced Th 17 dif-ferentiation.Neutralizing IL-22 in synovium of patients with RA might be an effective therapeutic strategy for RA treatment.
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Objective: To analyze the correlation between single nucleotide polymorphisms (SNP) of solute carrier family 22 member4 (SLC22A4) and runt-related transcription factor 1 (RUNX1) gene with rheumatoid arthritis (RA) and ankylosing spondylitis(AS) in Chinese Han ethnicity. Methods: Case-control studies were conducted with an RA cohort (104 RA patients and 109 healthy subjects) and an AS cohort (278 AS patients and 417 healthy controls). Three SNPs of SLC22A4 gene and an SNP of RUNX1 gene were genotyped by direct sequencing; we also assessed whether these alleles and genotypes were associated with RA and AS. Results: No significant differences in the distribution of the alleles and genotypes of SLC22A4 and RUNX1 polymorphisms were found between patients with RA and AS and healthy controls. Conclusion: Our results suggest that SLC22A4 and RUNX1 polymorphisms analyzed in the present study are not the susceptible genes for RA and AS in Chinese Han ethnicity.