Effect of continuous positive airway pressure treatment on Th17/Treg imbalance in patients with obstructive sleep apnea and a preliminary study on its mechanism.

BACKGROUND: Obstructive sleep apnea (OSA) can be considered a chronic inflammatory disease that impacts all bodily systems, including the immune system. This study aims to assess the Th17/Treg pattern in patients with OSA and the effect of continuous positive airway pressure (CPAP) treatment. METHODS: OSA patients and healthy controls were recruited. OSA patients recommended for CPAP treatment were followed up for three months. Flow cytometry was employed to determine the proportion of Th17 and Treg cells. Real-time quantitative polymerase chain reaction (PCR) and western blotting were utilized to detect the mRNA and protein levels of receptor-related orphan receptor γt (RORγt) and forkhead/winged helix transcription factor (Foxp3), respectively, in peripheral blood mononuclear cells (PBMCs). Enzyme-linked immunosorbent assay (ELISA) was performed to measure the serum levels of interleukin-17 (IL-17), IL-6, transforming growth factor-ß1 (TGF-ß1), and hypoxia-induced factor-1α (HIF-1α). RESULTS: A total of 56 OSA patients and 40 healthy controls were recruited. The proportion of Th17 cells, Th17/Treg ratio, mRNA and protein levels of RORγt, and serum IL-17, IL-6, and HIF-1α levels were higher in OSA patients. Conversely, the proportion of Treg cells, mRNA and protein levels of Foxp3, and serum TGF-ß1 levels were decreased in OSA patients. The proportion of Th17 and Treg cells in OSA can be predicted by the apnea hypopnea index (AHI), IL-6, TGF-ß1 and, HIF-1α. 30 moderate-to-severe OSA patients were adherent to three-month CPAP treatment, with improved Th17/Treg imbalance, IL-17, IL-6, TGF-ß1, and HIF-1α levels compared to pre-treatment values. CONCLUSION: There was a Th17/Treg imbalance in OSA patients. The prediction of Th17 and Treg cell proportions in OSA can be facilitated by AHI, as well as serum IL-6, TGF-ß1, and HIF-1α levels. Furthermore, CPAP treatment can potentially improve the Th17/Treg imbalance and reduce proinflammatory cytokines in OSA patients.

NF-κB-driven miR-34a impairs Treg/Th17 balance via targeting Foxp3.

The subset of regulatory T (Treg) cells, with its specific transcription Foxp3, is a unique cell type for the maintenance of immune homeostasis by controlling effector T (Teff) cell responses. Although it is common that a defect in Treg cells with Treg/Teff disorder causes autoimmune diseases; however, the precise mechanisms are not thoroughly revealed. Here, we report that miR-34a could attenuate human and murine Foxp3 gene expression via targeting their 3' untranslated regions (3' UTR). The human miR-34a, increased in peripheral blood mononuclear cells (PBMCs) and CD4+ T cells from rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) patients, displayed a positive correlation with some serum markers of inflammation including rheumatoid factor (RF), anti-streptolysin antibody (ASO), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) as well as Th17 signature gene RORγt, but inversely correlated with the mRNA expression levels of FOXP3. In addition, murine miR-34a levels were downregulated in TGF-ß-induced Treg cells but upregulated in Th17 cells induced in vitro compared to activated CD4+ T cells. It has also been demonstrated that elevated miR-34a disrupting Treg/Th17 balance in vivo contributed to the progress of pathogenesis of collagen induced arthritis (CIA) mice. Furthermore, IL-6 and TNF-α were responsible for the upregulation of miR-34a and downregulation of Foxp3, which was reverted by the addition of NF-κB/p65 inhibitor BAY11-7082, thus indicating that NF-κB/p65 inhibited Foxp3 expression in an miR-34a-dependent manner. Finally, IL-6 or TNF-α-activated p65 could bind to the miR-34a promotor and enhance its activity, resulting in upregulation of its transcription. Taken together, we show that NF-κB activated by inflammatory cytokines, such as IL-6 and TNF-α, ameliorates Foxp3 levels via regulating miR-34a expression, which provides a new mechanistic and therapeutic insight into the ongoing of autoimmune diseases.

Crying Time and RORγ/FOXP3 Expression in Lactobacillus reuteri DSM17938-Treated Infants with Colic: A Randomized Trial.

OBJECTIVES: To evaluate crying time, retinoid-related orphan receptor-γ (RORγ) and forkhead box P3 (FOXP3) messenger RNA levels (transcription factors that can modulate T cell responses to gut microbes), and to investigate gut microbiota and fecal calprotectin in infants treated with Lactobacillus reuteri for infantile colic. STUDY DESIGN: A double-blind, placebo-controlled randomized trial was conducted in primary care in Torino from August 1, 2015 to September 30, 2016. Patients suffering from infantile colic were randomly assigned to receive daily oral L reuteri (1 × 108 colony forming unit) or placebo for 1 month. Daily crying times were recorded in a structured diary. FOXP3 and RORγ messenger RNA in the peripheral blood was assessed with real-time TaqMan reverse transcription polymerase chain reaction. Gut microbiota and fecal calprotectin were evaluated. RESULTS: After infants with colic were supplemented with L reuteri DSM 17938 for 30 days, crying times were significantly shorter among infants with colic in the probiotic group compared with infants in the placebo group (74.67 ± 25.04 [IQR = 79] minutes /day vs 147.85 [IQR = 135] minutes /day [P = .001]). The FOXP3 concentration increased significantly (P = .009), resulting in decreased RORγ/FOXP3 ratios: 0.61 (IQR = 0.60) at day 0 and 0.48 (IQR = 0.28) at day 30 (P = .028). Furthermore, the probiotic increased the percentage of Lactobacillus (P = .049) and decreased fecal calprotectin (P = .0001). CONCLUSIONS: Infants with colic treated with L reuteri for 30 days had a significantly decreased crying time and an increased FOXP3 concentration, resulting in a decreased RORγ/FOXP3 ratio. The treatment reduced fecal calprotectin. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00893711.

Role of IL-6/RORC/IL-22 axis in driving Th17 pathway mediated immunopathogenesis of schizophrenia.

The immuno-inflammatory origin of schizophrenia in a subset of patients is viewed as a key element of an overarching etiological construct. Despite substantial research, the immune components exerting major effect are yet to be fully clarified. Disrupted T cell networks have consistently been linked to the pathogenesis of schizophrenia. Amongst the Th cell subsets, the Th17 cells have emerged as a paradigmatic lineage with significant functional implications in a vast number of immune mediated diseases including brain disorders such as schizophrenia. The present study was aimed at examining the functional role of the Th17 pathway in schizophrenia. To address this, genotyping of IL17A (rs2275913; G197A) Single Nucleotide Polymorphism was carried out by the PCR-RFLP method in 221 schizophrenia patients and 223 healthy control subjects. Gene expression of two transcription factors STAT3 and RORC was quantified in a subset of drug naïve schizophrenia patients (n = 56) and healthy controls (n = 52) by TaqMan assay. The plasma levels of fifteen cytokines belonging to Th17 pathway were estimated in a subset of drug naïve schizophrenia patients (n = 61) and healthy controls (n = 50) by using Bio-Plex Pro Human Th17 cytokine assays. The AA genotype was associated with higher total score of bizarre behaviour and apathy in female schizophrenia patients. A high gene expression level of RORC was observed in drug naïve schizophrenia patients. In addition, significantly elevated plasma levels of IL-6 and IL-22, and reduced levels of IL-1ß and IL-17F were noted in schizophrenia patients. Taken together, these findings indicate a dysregulated Th17 pathway in schizophrenia patients.

Increased Th17 functions are accompanied by Tregs activities in lupoid leishmaniasis.

The immunopathogenesis of lupoid leishmaniasis is challenging. Although an appropriate immune response is critical for controlling these parasites, inappropriate inflammatory reactions can also promote increased pathology. The role of immune modulatory effect of the main transcription factors and cytokines of T regulatory and Th17 cells in pathogenesis of leishmaniasis chronicity was investigated in this study. The gene expression of interleukin-10 (IL-10), transforming growth factor-ß (TGF-ß1), forkhead box P3 (Foxp3), interleukin-17(IL-17A) and retinoic acid-related orphan receptor gamma t (ROrC) was assessed in peripheral blood mononuclear cells of eighty blood samples from cutaneous leishmaniasis (CL) patients with usual lesions (n = 31), lupoid lesions (n = 29) and healthy volunteers (n = 20). Quantitative relative real-time PCR (qRT-PCR) was performed using the Taqman and Sybergreen methods for expression of target genes. Expression of Foxp3 (P = .013), IL-10 (P < .001) and IL-17A (P < .001) was significantly higher in lupoid patient compare to the nonlupoid group. Expression of Foxp3 (P < .001), IL-10 (P < .001) and IL-17A (P = .033) was significantly more in nonlupoid subjects than in healthy volunteers, except for RORγt. These findings suggest that Foxp3+ cells, IL-10 and IL-17 play important roles in the immunopathogenesis of CL and that these roles differ depending on the causal leishmania species and different body compartments.

Association between cytokine profile and transcription factors produced by T-cell subsets in early- and late-onset pre-eclampsia.

Pre-eclampsia (PE) is an obstetric pathology characterized by abnormal activation of the innate and adaptive immune systems dependent on the imbalance of T helper subsets. The present study aimed to evaluate the gene and protein expression of T helper type 1 (Th1)/Th2/Th17/regulatory T (Treg) cell transcription factors in peripheral blood lymphocytes from pregnant women with PE employing quantitative RT-PCR and flow cytometry techniques, as well as the cytokine profile produced by these CD4+ T-cell subsets in the plasma of pregnant women with PE, classified as early-onset PE (n = 20), late-onset PE (n = 20) and normotensive pregnant women (n = 20). Results showed a higher percentage of CD4+ T cells expressing the RORc transcription factor (Th17) and a lower percentage of cells expressing FoxP3 (Treg) in women with early-onset PE compared with late-onset PE and normotensive groups. A lower gene expression of GATA-3 transcription factor was detected in cells of women with early-onset PE compared with the late-onset PE group. Endogenous plasma levels of interleukin-6 (IL-6), IL-17 and tumour necrosis factor-α were significantly higher in the early-onset PE group than in the late-onset PE and normotensive groups, whereas IL-4 (Th2 profile) and IL-22 (Th17 profile), were not significantly different between the studied groups. The endogenous levels of transforming growth factor-ß and IL-10 were significantly lower in the pre-eclamptic than in the normotensive groups of the same gestational age, with a significant difference between early- and late-onset PE. The results show that in women with PE there is an imbalance between inflammatory and anti-inflammatory profiles in CD4+ T-cell subsets, with polarization to Th17 profiles in the early-onset PE, considered as the severe form of PE.

The effects of T helper 17 and regulatory T cells on patients with carotid atherosclerosis.

This study was aimed to observe the level of T helper 17 (Th17) cells and CD4+ CD25+ Foxp3+ regulatory T cells (Tregs) and their related factors in carotid atherosclerosis (AS) patients and AS model rats to explore the influence of Th17 on the pathological process of AS and its specific mechanism. 60 cases with AS in our hospital from July 2012 to July 2014 were recruited for this study as the observation group, and 40 healthy people who came to the hospital for a physical examination were the control group. Flow cytometry (FCM) was used to detect the Th17 and Treg cells in the peripheral blood of the two groups, ELISA was used to detect IL-17 and transforming growth factor-ß (TGF-ß), and RT-PCR was used to test the RORγT mRNA and Foxp3 mRNA expression levels. An AS model was created in rats using high-fat+ VD3 to explore the mechanism of Th17 on AS. The Th17 count, serum level of IL-17, and RORγT mRNA level of the observation group were significantly higher than those of the control group (P<0.001). The Tregs count, serum TGF-ß level, and Foxp3 mRNA level were significantly lower in the observation group than in the control group (P<0.001). In addition, the findings of the AS model in rats showed that the Th17 cell count and serum level of IL-17 in high-fat rats were significantly higher than in the normal rats (P<0.05). The Treg count and TGF-ß levels of the observation rats were significantly lower than in the normal rats (P<0.05). The IL-17 level, serum total cholesterol, and triglyceride in the high-fat-feed rats decreased after being injected with the IL-17 neutralizing antibody, but TGF-ß levels increased, and the difference was significant (P<0.05). Th17 cells and their related factors can be involved in promoting the pathological process of AS, while Tregs and its related factors can be involved in the inhibition of AS. Blocking IL-17 can be one potential method of treating AS.

Down-regulation of expression of retinoid acid-related orphan receptor C (RORC) in systemic lupus erythematosus.

CONTEXT: Retinoic acid-related orphan receptor C (RORC), the key factor orchestrating the transcription of genes encoding interleukin 17, plays a critical role in the regulation of inflammatory responses. OBJECTIVE: The objective of this study was to analyze the expression of RORC in the peripheral blood of patients with systemic lupus erythematosus (SLE) for a better understanding of the pathogenesis of SLE especially in relation to disease activity and clinical and biochemical findings. METHODS: The study included 24 patients with SLE and a control group of 18 healthy gender- and age-matched individuals. Evaluation of the level of expression of RORC mRNA was performed by real-time polymerase chain reaction. RESULTS: The results showed that patients with SLE had lower RORC gene expression levels compared with healthy subjects that were not correlated with disease activity. The down-regulation of RORC was significantly lower in patients with lupus nephritis in remission than active lupus nephritis and nonrenal patients. CONCLUSIONS: The findings suggest that RORC plays a significant role in the dysregulated immune response associated with SLE. Deciphering the intricate regulatory network and the target genes of RORC will help unravel new specific treatments for SLE.

Upregulated Cellular Expression of IL-17 by CD4+ T-Cells in Osteoporotic Postmenopausal Women.

AIM: The study aimed to investigate cellular expression of IL-17 by CD4+ T-cells in osteoporotic postmenopausal women. METHODS: We enrolled 25 postmenopausal women with osteoporosis (PostMO) and 25 postmenopausal women with normal bone mineral density measurements (PostM) to examine the production of IL-17, tumor necrosis factor α (TNFα) and receptor activator of nuclear factor x03BA;-B ligand (RANKL) by CD4+ T-cells and IL-17, RORx03B3;t, TNFα and RANKL mRNA levels in CD4+ T-cells. Circulating concentrations of IL-17 along with IL-6, TNFα, RANKL and osteoprotegerin (OPG) were also determined. RESULTS: Osteoporotic postmenopausal women had higher serum concentrations of IL-17 (3.7 ± 1.3 vs. 2.5 ± 1.1 ng/ml, p = 0.042), IL-6 (158 ± 56 vs. 105 ± 39 pg/ml, p = 0.044), TNFα (138 ± 41 vs. 74 ± 11 pg/ml, p < 0.001) and OPG (1.7 ± 0.4 vs. 1.3 ± 0.4 ng/ml, p = 0.039) than healthy controls. The IL-17-producing CD4+ T-cells were higher in the PostMO group than in the PostM group (7.1 ± 2.4 vs. 4.9 ± 1.4%, p = 0.0015). Additionally, osteoporotic postmenopausal women had greater mRNA levels of IL-17 (3.5 ± 2.9 vs. 1.2 ± 1.0%, p = 0.019) and RORx03B3;t (5.7 ± 2.5 vs. 2.2 ± 1.0%, p < 0.001) in CD4+ T-cells than in healthy controls. CONCLUSIONS: Our findings implied that the upregulated production of IL-17 may play an important role in regulating bone loss in osteoporotic postmenopausal women.

Expression of retinoid-related orphan receptor (ROR)γt on NK22 cells in the peripheral blood and uterine endometrium of women with unexplained recurrent pregnancy loss and unexplained infertility.

AIM: Recently, NK22 cells, a subset of interleukin (IL)-22-producing natural killer (NK) cells, were identified. We have previously reported the higher percentage of NK22 cells in women suffering recurrent pregnancy loss (RPL). Moreover, we have also reported lower expression of NKp46, a kind of natural cytotoxicity receptor (NCR), on NK cells and the changes of NK cell producing cytokines in women who experience RPL. NK22 cells express NCRs, such as NKp44 or NKp46. Retinoid-related orphan receptor γt (RORγt) is known as a regulator of NK22 cells; however, in NK22 cells of peripheral blood (PB) and the uterine endometrium (UE), the relationship between NCRs and RORγt is unclear. We investigate RORγt expression NK22 cells in the PB and UE of women with unexplained infertility (uI) or unexplained RPL (uRPL). METHODS: Lymphocytes were extracted from PB and UE, derived from women with uI or uRPL. Expression of RORγt and NCRs in NK cells and NK cell-produced cytokines were analyzed by flow cytometry. RESULTS: CD56+ /NKp46+ /RORγt+ cells were positively correlated with CD56+ /IL-22+ cells in both PB and UE. CD56bright /NKp46bright /RORγt+ cells were significantly higher in uRPL than in uI, and endometrial CD56bright /NKp46bright /RORγt+ cells were positively correlated with PB. In UE, CD56bright /RORγt+ cells were negatively correlated with CD56bright /interferon-γ+ and CD56bright /tumor necrosis factor-α+ cells of uRPL. CONCLUSION: RORγt may be associated with NK22 cells in reproduction. Particularly, higher expression of RORγt may be associated with elevated NK22 cells in uRPL.

RORC2 Genetic Variants and Serum Levels in Patients with Rheumatoid Arthritis.

BACKGROUND: In the present study, we aimed to evaluate whether polymorphisms within the RORc2 gene are involved in the risk and severity of rheumatoid arthritis (RA). METHODS: 591 RA patients and 341 healthy individuals were examined for RORc2 gene polymorphisms. Serum retinoic acid receptor-related orphan receptor C (RORc) levels were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The rs9826 A/G, rs12045886 T/C and rs9017 G/A RORc2 gene SNPs show no significant differences in the proportion of cases and control. Overall, rs9826 and rs9017 were in high linkage disequilibrium (LD) with D' = 0.952 and r² = 0.874, except rs9826 and rs12045886; and rs12045886 and rs9017 in weak LD. The genotype-phenotype analysis showed a significant association between RORc2 rs9826 A/G and rs9017 G/A single nucleotide polymorphisms (SNPs) and median of C-reactive protein (CRP). Serum RORc levels was higher in RA patients with rs9826AA, rs12045886TT and -TC, and rs9017AA genotypes compared to healthy subjects with the same genotypes (p = 0.02, p = 0.04 and p = 0.01, respectively). Moreover, the median of RORc protein level was higher in RA patients with number of swollen joints bigger then 3 (p = 0.04) and with Health Assessment Questionnaires (HAQ) score bigger then 1.5 (0.049). CONCLUSIONS: Current findings indicated that the RORc2 genetic polymorphism and the RORc2 protein level may be associated with severity of RA in the Polish population.

Expression of Th17 and CD4+ CD25+ T regulatory cells in peripheral blood of acute leukemia patients and their prognostic significance.

To discuss the expression of T helper cell 17 (Th17) cells and CD4+ CD25+ Foxp3+ regulatory T cells (Treg) in peripheral blood (PB) of patients with acute leukemia (AL), and to explore the relationship between them and disease prognosis. 40 patients diagnosed with acute leukemia in The First Affiliated Hospital of Zhengzhou University from July 2012 to August 2014 were selected as the observation group. Meanwhile, 40 healthy people were taken as the control group. Flow Cytometry Method (FCM) was used to detect the level of Th17 cells and CD4+ CD25+ Foxp3+ cells in peripheral blood of the two groups, and enzyme-linked immuno sorbent assay (ELISA) method was used to test the level of IL17 and TGF-ß in peripheral blood of two groups; reverse transcription-polymerase chain reaction (RT-PCR) was adopted to analyze the mRNA levels of RORγT and Foxp3 in peripheral blood. In addition, we examined the levels of Th17 and CD4+ CD25+ Foxp3+ cells and associated factor levels in patients with remission after AL chemotherapy. the Th17 cells (CD3+ CD4+ IL-17+) in acute leukemia patients accounted for (1.51±0.27)%, which was significantly higher than that of control group (0.36±0.23)%, with statistical significance (t=20.51, P<0.001); the percentage of CD4+ CD25+ Foxp3+ cells in AL patients was (3.37±0.48)%, which was significantly higher than that of control group of (1.26±0.27)%, with statistical significance (t=24.23, P<0.001); the serum levels of IL-17 and TGF-ß in AL patients were (28.12±6.33) pg/ml and (38.41±8.44) pg/ml respectively, which were all significantly higher than that of control group of (14.41±6.21) pg/ml and (24.49±7.42) pg/ml, with statistical significance (t=7.83, P<0.001; t=7.83, P<0.001); the RORγT mRNA and Foxp3 mRNA levels in AL patients were all significantly higher than that of control group, with statistical significance (t=12.27, P<0.001; t=7.89, P<0.001). In addition, compared with before chemotherapy, the levels of Th17 cells and CD4+ CD25+ Foxp3+ cells, and the serum levels of IL-17 and TGF-ß in acute leukemia patients all decreased significantly after 6 months of chemotherapy, and the difference was statistically significant (P<0.001). Th17 cells, CD4+ CD25+ Foxp3+ cells and their secretory proteins IL-17, TGF-ß and transcription factors were significantly increased in AL patients. Therefore, regular detection of peripheral blood Th17 and Treg cells, as well as their secretory proteins are useful for monitoring the immune status and prognosis of patients.

Basal level of Th17 immune response is not enhanced in aplastic amemia.

Acquired aplastic anemia (AA) is an immune mediated bone marrow failure syndrome which is associated with impaired T-cell immune responses. In this work we investigated the role of Th17 immune response in AA. Our results showed that the absolute numbers of circulating IL-17-producing CD4(+) T cells and CD4(+)CD161(+)CCR6(+) cells, the blood plasma level of IL-17, and the expression level of Th17 lineage-specifying transcription factor RORC in circulating CD4(+) T cells, were not increased in AA. These results suggest that Th17 immune response may not play an important role in the pathogenesis of AA.

The existence of Th22, pure Th17 and Th1 cells in CIN and Cervical Cancer along with their frequency variation in different stages of cervical cancer.

BACKGROUND: Recently, it is found that T-helper (Th) 22 cells are involved in different types of autoimmune and tumor diseases. But, till now, no study has been carried out to understand the involvement of these cells in cervical cancer (CC). METHODS: Flow cytometry was used to determine the expression of interferon gamma (IFN-γ), Interleukin-22 (IL-22), IL-17 in the peripheral blood of healthy controls (HC), CIN and cervical cancer patients. From peripheral blood mononuclear cells (PBMCs), mRNA expression levels of Aryl hydrocarbon receptor (AHR), RAR-related orphan receptor C (RORC), TNF-α and IL-6 were respectively determined. Using the method of ELISA, plasma concentrations of IL-22, IL-17 and TNF-α were examined. RESULTS: Th22 and Th17 cells were elevated in CC and CIN patients. Th1 cells and the plasma concentrations of IL-22 in CC patients were significantly increased compared with HC. In CC patients, an increased prevalence of Th22 cells was associated with lymph node metastases. There was a positive correlation between Th22 and Th17 cells, but an approximately negative correlation between Th22 and Th1 cells in CC patients. The mRNA expression of RORC, TNF-α and IL-6 was significantly high in CC patients. CONCLUSIONS: Our results indicate that there is a higher circulatory frequency of Th22, Th17 and Th1 cells in CC which may conjointly participate in the pathogenesis and growth of CC.

Increased interleukin-27 promotes Th1 differentiation in patients with chronic immune thrombocytopenia.

Chronic immune thrombocytopenia (cITP) is an autoimmune disease with disturbed cytokine profile. Although plasma levels of IL-27 are shown to be associated with cITP, its association with T cell subsets has not been studied. The objective of this study was to study the association between IL-27 and different T cell subsets in patients with cITP. Heparinized blood was collected from 31 patients with cITP and 36 healthy controls (platelet count <100 × 10(9)/l and 103-280 × 10(9)/l, respectively). The percentage of Th1, Th2 and Th17 cells in peripheral blood mononuclear cells (PBMCs) were enumerated by flow cytometry, and the mRNA levels of IL-27, T-bet, GATA-3 and retinoid-related orphan receptor gamma (RORγt) by real-time reverse transcriptase polymerase chain (RT-PCR). Plasma cytokine levels of IL-27, interferon-gamma (IFN-γ), IL-4 and IL-17A were estimated by flow cytometrix. The effect of exogenous recombinant IL-27(rhIL-27) on the differentiation of T cells into Th1, Th2 and Th17 cells was investigated by cell culture. The percentage of Th1 and Th17 cells and the plasma concentration and mRNA levels of IL-27 were significantly higher in cITP patients compared with healthy controls. Plasma levels of IL-27 correlated positively with percentage of Th1 cells in patients with cITP. Exogenous (rhIL-27) could significantly up-regulate the percentage of Th1 cells and down-regulate Th2 cells in vitro. Th17 cells were reduced in the presence of (rhIL-27) in controls but had no effect in patients with cITP. The up-regulation of IL-27 might cause Th1 differentiation and might be involved in the pathophysiology of cITP.

Alteration of Th17 and Treg cells in patients with unexplained recurrent spontaneous abortion before and after lymphocyte immunization therapy.

BACKGROUND: Several types of T cells have been associated with the pathogenesis of unexplained recurrent spontaneous abortion (URSA), including Th1/Th2/Th17/Tregs cell. It has been appreciated that immunotherapy with paternal or third party lymphocytes is an effective method of treatment for URSA patients. The balance of Th1/Th2 cells could be maintained and an increase of Treg cells would be beneficial after immunotherapy; however, the mechanism by which the Th17/Treg balance affects URSA has not yet been fully elucidated. METHODS: Here, we used flow cytometry, liquid chip technology and quantitative real-time PCR (qPCR) methods to characterize Th17/Treg cell populations after immunotherapy. We found that after immunotherapy in URSA patients, the percentage of Th17 cells decreased and the percentage of Treg cells in peripheral blood mononuclear cells (PBMC) increased, as detected by flow cytometry. RESULTS: Immunotherapy may induce a decrease in the Th17/Treg ratio and the Treg bias, which may be beneficial for the maintenance of pregnancy. The expression level of ROR gamma t, a transcription factor found in Th17 cells, decreased and the expression of the Treg-specific transcription factor Foxp3 increased in peripheral blood as detected by qPCR. Immunotherapy may induce a decrease in the ratio of ROR gamma t to Foxp3 and a Treg cell bias, which would be beneficial for pregnancy maintenance. The secretion of the Treg-associated cytokine TGF-beta, as well as Th2 cytokines, was increased in serum, while the secretion of Th17-associated cytokine IL-17A and Th1 cytokine production was decreased. The Th1/Th2 cytokine ratio significantly decreased. Similarly, the Th17/Treg ratio significantly decreased in the total patient after immunotherapy. CONCLUSIONS: These results indicate that in patients with URSA, immunotherapy with mononuclear cells derived from the baby's father could affect both Th1/Th2 and Th17/Treg balance, and we found that the Th2 and Treg bias would be beneficial for pregnancy, which may lead to a balancing of the Th17/Treg ratio in URSA patients after immunotherapy.

Effects of probiotic yogurt on fat distribution and gene expression of proinflammatory factors in peripheral blood mononuclear cells in overweight and obese people with or without weight-loss diet.

OBJECTIVES: The purpose of this study was to investigate whether probiotics had an effect on proinflammatory markers and cytokines in overweight and obese individuals and whether they could have synergistic effects with weight-loss diets. METHODS: A total of 75 healthy overweight and obese individuals completed this randomized doubled-blind controlled clinical trial. Participants were randomly assigned to groups consuming regular yogurt with a low-calorie diet (LCD, RLCD; n = 25) or receiving probiotic yogurt with LCD (PLCD; n = 25) or consuming probiotic yogurt without LCD (PWLCD; n = 25) for 8 weeks. The pribiotic regimen contained 200 g/day yogurt, enriched by Lactobacillus acidophilus La5, Bifidobacterium BB12, and Lactobacillus casei DN001 10(8) colony-forming units/g. Body fat percentage, high-sensitive C-reactive protein (hs-CRP), tumor necrosis factor-alpha (TNF-α), leptin, and mRNA levels of inflammation-related genes (TNF-α and RAR-related orphan receptor gamma [ROR-γt]) in peripheral blood mononuclear cells (PBMCs) were measured. RESULTS: A reduction in body mass index (BMI), fat percentage, and leptin level was observed that was more obvious in groups who received the weight-loss diet with probiotic yogurt. Reduction in the gene expression of ROR-γt was significant in the PLCD group (p < 0.001). The expression of TNF-α did not change among all groups after intervention. The mean concentration of leptin was significantly decreased in all groups after the dietary intervention, but the mean changes in leptin level in the PLCD group was more prominent compared to the other two groups (-2.38, p < 0.001 [PLCD] vs -1.75, p = 0.002 [RLCD] and -0.55 ng/mL, p = 0.12 [PWLCD]). The reduction in serum levels of hs-CRP was more evident in the PWLCD group compared to the PLCD and RLCD groups after the 8-week intervention (-3.4, p = 0.03 vs -1.76, p < 0.001 and -2.98 pg/mL, p < 0.001, respectively). CONCLUSION: Our results suggested that the weight-loss diet and probiotic yogurt had synergistic effects on T-cells subset specific gene expression in PBMCs, fat percentage, and body weight among overweight and obese individuals.

Increased frequency of Th17 cells in the peripheral blood of children infected with enterovirus 71.

Enterovirus 71 (EV71) affects the health of young children globally causing severe neurologic diseases. The relationship between EV71 infection and T helper type 17 (Th17) has not been described, although this new Th subset or interleukin-17 (IL-17) has been reported to be associated with other viral infections. The purpose of the current study was to describe the immune profile involving Th17 cells, neutrophils, and related factors and to speculate on the possible immunopathogenesis of EV71 infections. Flow cytometry and an automatic blood cell counter were used to analyze circulating Th17 cells and count neutrophils, respectively. Expression of acid-related orphan nuclear receptor gamma t (ROR γt) was evaluated by reverse-transcriptional PCR, and enzyme linked immunosorbent assays (ELISAs) were used for detecting concentrations of IL-17, IL-23, and IFN-γ. The results showed that the frequencies of Th17 cells (1.47 ± 0.87%) and the number of neutrophils (7.4 ± 4.1 × 10(9) /L) in peripheral blood samples from children infected with EV71 were significantly higher compared to controls. In addition, there was a statistically higher expression of ROR γt in peripheral blood mononuclear cells (PBMCs) and elevated concentrations of IL-17 and IL-23 in sera, but lower IFN-γ production during EV71 infections. The findings suggest that Th17 cells are mediators during the immunologic process.

Th17/Treg imbalance in adult patients with minimal change nephrotic syndrome.

To determine whether Th17/Treg balance was abnormal in adult patients with minimal change nephrotic syndrome (MCNS), we studied 25 patients with new-onset MCNS and 20 normal persons. The results showed that MCNS patients exhibited a significant increase in Th17 number, Th17-related cytokines (IL-17 and IL-23), and transcription factor (RORγt) levels, as well as an obvious decrease in Treg number, Treg-related cytokines (TGF-ß1 and IL-10), and transcription factor (Foxp3) levels. The Th17/Treg ratios increased along with increased proteinuria and decreased albumin levels in patients with MCNS. IL-17 protein expression was also detected in the renal biopsy tissue of MCNS patients, particularly in patients with acute renal failure. Further, Th17/Treg balance returned to normal after effective corticosteroids therapy in 16 MCNS patients. These results indicated that Th17/Treg imbalance existed in MCNS patients, suggesting a potential role of Th17/Treg imbalance in the pathogenesis of MCNS.

Decreased expression of interleukin-27 in immune thrombocytopenia.

Primary immune thrombocytopenia (ITP) is an immune-mediated disorder in which disturbed cytokine profiles have been found. Interleukin-27 (IL27) has been shown to bear both proinflammatory and anti-inflammtory effects. In the present study, plasma levels of IL27, interferon gamma (IFNG), IL4, and IL17A were determined by enzyme-linked immunosorbent assay in 23 active ITP patients, 20 patients in remission and 20 healthy controls. mRNA expression levels of IL27, EBI3, IL27 receptor (IL27RA), IL17A and RAR-related orphan receptor C (RORC) were determined by real-time quantitative polymerase chain reaction. Significantly lower levels of plasma IL27, IL4, mRNA expression of IL27, EBI3 and higher levels of plasma IFNG as well as mRNA expression of IL17A, RORC were observed in active ITP patients compared with healthy controls or patients in remission. No statistical difference was found in IL27RA mRNA expression or plasma IL17A levels among active ITP patients and controls. A negative correlation was found between the IL27 and RORC mRNA expression levels in active ITP patients. Our data demonstrated that active ITP patients had decreased plasma and mRNA expression levels of IL27, suggesting that it might be involved in the pathophysiological process of ITP.