Disturbed Th17/Treg balance in patients with rheumatoid arthritis.

Proinflammatory Th17 cells and CD4(+)CD25(+) regulatory T (Treg) cells are two newly identified T lymphocyte subsets, which have opposite effects on autoimmunity and inflammation. To assess the Th17/Treg pattern and cytokine microenvironment in peripheral blood of patients with RA, we included 66 RA patients and 20 healthy volunteers. Of all these subjects, peripheral Th17 and Treg frequencies were analyzed by flow cytometry (FCM) and the plasma levels of interleukin (IL)-17, 23, 6, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-ß1 were detected by ELISA. The results demonstrated that RA patients revealed an obvious increase in peripheral Th17 frequencies and levels of Th17-related cytokines (IL-17, IL-23, IL-6, TNF-α) while a significant decrease in Treg frequencies and Treg-related cytokine (TGF-ß1) levels when compared with healthy people. Our study indicated that development of RA is associated with peripheral Th17/Treg imbalance and characterized by a proinflammatory cytokine microenvironment, which supports continuing generation of Th17 cells.

Association study of single nucleotide polymorphisms in pre-miRNA and rheumatoid arthritis in a Han Chinese population.

The aim of this study was to perform an association study between two single nucleotide polymorphisms (SNPs) rs2910164 G>C and rs3746444 T>C in pre-miRNA (hsa-mir-146a and hsa-mir-499) and rheumatoid arthritis (RA) in the Han Chinese population. 208 Han Chinese patients with RA and 240 healthy controls were recruited in this study. The SNPs was genotyped by polymerase chain reaction-restriction fragment length polymorphism. Anti-cyclic citrullinated peptide (anti-CCP) antibody was measured by enzyme linked immunosorbent assay and rheumatoid factor (RF) was measured by rate nephelometry. The genotype frequencies between cases and controls were compared by χ(2) analysis. No significant association between the SNPs (rs2910164 and rs3746444) and RA was observed (P = 0.631 and 0.775, respectively), and the SNPs did not show any association with the RF-positive (P = 0.631 and 0.775, respectively). However, there was a significant difference on the level of anti-CCP antibody between different genotypes in rs3746444 (P = 0.007). The heterozygote CT had significantly higher level of anti-CCP antibody compared with homozygote CC and TT (P = 0.054 and 0.003, respectively). We first investigated the association between the SNPs (rs2910164 G>C and rs3746444 T>C) in the pre-miRNA (hsa-mir-146a and hsa-mir-499) and RA in a Han Chinese population. We did not find a significant association between the SNPs and the susceptibility to RA, while the SNP rs3746444 may affect anti-CCP antibody production.

[Promising diagnostic model for systemic lupus erythematosus using proteomic fingerprint technology].

OBJECTIVE: To establish a diagnostic model for systemic lupus erythematosus (SLE) using proteiomic fingerprint techology. METHODS: Blood samples were collected from 64 cases of SLE, 30 cases of rheumatoid arthritis (RA), 30 cases of Sjogren's syndrome (SS), 25 cases of systemic sclerosis (SSc), as well as 83 healthy controls. Proteomic spectra of these 232 serum samples were generated by proteomic fingerprint technology. Diagnostic model was established by a machine learning algorithm called decision boosting. The sensitivity and specificity of the diagnostic model was validated with a blinded testing set. RESULTS: Sixty differential protein peaks (P<0.05) between SLE and control subjects were indicated, 28 of them were up regulated and 32 were down regulated in SLE patients. The algorithm identified a cluster pattern segregating SLE from non-SLE with sensitivity of 91% and specificity of 92%. The discriminatory diagnostic pattern correctly identified SLE. A sensitivity of 78% and specificity of 96% for the blinded test were obtained when comparing SLE vs non-SLE. CONCLUSION: This diagnostic model using proteiomic fingerprint techology appears to be a promising tools with high sensitivity and specificity in diagnosis of SLE.

[Study on proteomics of familial systemic lupus erythematosus patients in one family from Sichuan, China].

OBJECTIVE: To investigate the proteomic characteristics of systemic lupus erythematosus (SLE) in a SLE family from Sichuan, China which consisting of 7 members with 3 SLE cases, and to find the proteins correlated with the heredity of SLE. METHODS: A total of 153 serum samples were collected from 7 members including 3 SLE sisters in this SLE family, 63 individual SLE patients, as well as 83 healthy controls. The diagnosis of SLE is based on the American College of Rheumatology criteria (1997). All serum samples were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) combined with magnetic beads technology. Serum protein profiles were obtained by MALDI-TOF-MS combined with magnetic beads in order to identify predictive biomarkers of risk of suffering SLE. The resulting spectra were analyzed with Biomarker Wizard software 3.1.0. RESULTS: Four discriminative mass/charge (m/z) proteins serving as pathogenic biomarkers were identified on arrays for family SLE cases versus individual SLE and healthy controls. The protein level of peak intensities at m/z of 9342.23 was significantly greater in SLE family group compared with that in individual SLE patients and healthy controls (P<0.05), those of individual SLE patients were significantly greater compared with healthy controls (P<0.05); the proteins level of peak intensities at m/z of 4094.03, 5905.35 and 7973.53 in SLE family group were significantly lower compared with that in individual SLE patients and healthy controls (P<0.05), those of individual SLE patients were significantly lower compared with healthy controls (P<0.05). CONCLUSION: The proteins of m/z of 9342.23, 4094.03, 5905.35 and 7973.53 maybe play a great role in assemble pathogenesis of SLE and predict the risk of suffering SLE. The higher protein level of m/z of 9342.23 and the lower protein level of m/z of 4094.03, 5905.35 and 7973.53, the higher risk of sufferring with SLE.

Enhanced IL-6/phosphorylated STAT3 signaling is related to the imbalance of circulating T follicular helper/T follicular regulatory cells in patients with rheumatoid arthritis.

BACKGROUND: Follicular helper T (Tfh) cells are specialized in helping B lymphocytes, which play a central role in autoimmune diseases that have a major B cell component, such as in rheumatoid arthritis (RA). Follicular regulatory T (Tfr) cells control the over-activation of Tfh and B cells in germinal centers. Dysregulation of Tfh cells and Tfr cells has been reported to be involved in the pathogenesis of some autoimmune diseases. However, the balance of Tfh and Tfr cells, and their roles in the development and progression of RA are still not clear. METHODS: In this study, we enrolled 44 patients with RA (20 patients with active RA and 24 patients with inactive RA) and 20 healthy controls, and analyzed the frequencies of circulating Tfh and Tfr cells, expression of programmed death-1 (PD-1), inducible co-stimulator (ICOS), intracellular IL-21, and pSTAT3 in Tfh cells, and serum levels of IL-6. The correlation among these parameters and that of Tfh or Tfr cells with disease activity were also analyzed. RESULTS: Patients with RA (especially active RA) had higher frequencies of Tfh cells, but lower percentages of Tfr cells, thereby resulting in elevated ratios of Tfh/Tfr. Expression levels of PD-1 and IL-21 in Tfh cells were higher in patients with RA than in healthy subjects, while no difference in ICOS expression was observed between patients and controls. Both pSTAT3 expression and serum IL-6 levels increased in patients with RA, and positive correlation between them was observed. Additionally, pSTAT3 expression was positively correlated with Tfh cell frequency. The Disease Activity Score in 28 joints based on C-reactive protein (DAS28-CRP) was negatively correlated with Tfr cell frequency, but was positively correlated with both Tfh/Tfr ratio and PD-1 expression. CONCLUSIONS: Results demonstrated that enhanced IL-6/pSTAT3 signaling may contribute to promotion of Tfh cells, consequently skewing the ratio of Tfh to Tfr cells, which may be crucial for disease progression in RA.

[Analysis of frequency of peripheral blood CD4+; CD25(high);Tregs and CD4+; CD25(low);T cells and expression of PD-1 in SLE and RA patients].

AIM: To explore the frequencies of peripheral blood CD4(+);CD25(high);Treg and CD4(+);CD25(low);T cells and the expression of the co-stimulatory molecule PD-1 on these two group cells surface in SLE and RA patients, and to explore their roles in cell immunity disorder of SLE and RA. METHODS: Flow cytometry (FCM) was used to determine the frequencies of peripheral blood CD4(+);CD25(high);Treg and CD4(+);CD25(low);T cells, and the expression percentage of PD-1. RESULTS: Compared with healthy control, the frequencies of peripheral blood CD4(+);CD25(high);Treg from both SLE and RA patients groups decreased significantly(P<0.05). Compared between two disease groups, the frequency of peripheral blood CD4(+);CD25(high);Treg in SLE patients was significantly lower(P<0.05). The expression percentage of PD-1 on CD4(+);CD25(high);Treg surface in RA group was obviously lower than that in both healthy control and SLE patients groups(P<0.05), while the percentage had no significant difference between SLE patients and healthy control(P>0.05). There was no significant difference in both the frequency of CD4(+);CD25(low);T cells and the expression percentage of PD-1 on this subset cells among three groups. CONCLUSION: The weakened ability of peripheral blood CD4(+);CD25(high);Treg to suppress effector T cells resulted from their production deficiency is the common characteristic of SLE and RA patients. Decreased expression of PD-1 is the primary cause leading to the suppressive function of peripheral blood CD4(+);CD25(high);Treg weakened in RA patients. However, PD-1 does not play major role in weakening the suppression activity of CD4(+);CD25(high);Treg from SLE patients.

[Study on ratio imbalance of peripheral blood Th17/Treg cells in patients with rheumatoid arthritis].

AIM: To observe the relationship between balance of peripheral blood Th17 cells and Foxp3(+) CD4(+) CD25(+) regulatory T (Treg) cells in patients with rheumatoid arthritis (RA), and to clarify the role the ratio imbalance of peripheral blood Th17/Treg cells playing in pathogenesis of RA. METHODS: The ratio of peripheral blood Th17 cells and Foxp3(+) CD4(+) CD25(+) Treg cells in RA patients and healthy subjects were determined by flow cytometry (FCM). RESULTS: Compared with healthy controls, the ratio of both CD3(+) CD4(+) T cells and Th17 cells in RA patients increased significantly (P<0.05), while the percentage of Foxp3(+) CD4(+) CD25(+) Treg cells was markedly lower (P<0.05). With the development of RA activity, the ratio of Th17 cells increased (P<0.05), and the ratio of Foxp3(+) CD4(+) CD25(+) Treg cells decreased (P>0.05). CONCLUSION: The disorder of peripheral blood T lymphocyte subsets in RA patients characterized by increased CD4(+) T cells. The imbalance between Th17 cells and Foxp3(+) CD4(+) CD25(+) Treg cells resulted from increased ratio of Th17 cells and decreased ratio of Foxp3(+) CD4(+) CD25(+) Treg cells may play a critical role in RA progression.