Th17 Cells In Blood Research Articles

Molecular mechanisms of interleukin-38 inhibiting inflammatory bowel disease in children by regulating nuclear factor-κB and signal transduction and activator of transcription 3 pathway

Objective To explore the role of interleukin (IL)-38 in inhibiting inflammatory bowel disease (IBD) in children and to investigate the potential molecular mechanisms. Methods From January 2014 to October 2017, 67 patients with ulcerative colitis (UC) and 115 patients with Crohn′s disease (CD) admitted to Wuhan Children′s Hospital were recruited, and 40 individuals with normal endoscopic findings were selected as control. Serum levels of IL-38 of IBD patients and healthy control were determined by enzyme-linked immunosorbent assay (ELISA). Immunohistochemical staining (IHC) was used to detect the expression level of IL-38, nuclear factor κB (NF-κB), phosphorylated signal transduction and activator of transcription 3 (p-STAT3), C-reaction protein (CRP) and erythrocyte sedimentation rate (ESR) in the intestinal mucosa of IBD patients and healthy controls. The extent of disease, therapeutic agents and disease activity scores (Mayo score system for UC patients, Crohn′s disease activity index (CDAI) for CD patients) were evaluated. IL-38-C57BL/6 transgenic mice model was established, and dextran sulfate sodium was used to induce IBD mice model. The intestinal inflammation levels were compared between the wild type IBD mice and IL-38 transgenic IBD mice. The levels of IL-38, NF-κB and p-STAT3 in intestinal mucosa of mice of different groups were determined by IHC. The ratio of CD4+ IL-17+ T helper (Th) 17 cells in peripheral blood of mice of different groups was detected by flow cytometry. Independent sample t test, chi square test and Pearson correlation were performed for statistical analysis. Results The results of ELISA showed that the serum levels of IL-38 of UC and CD patients were (6.1±1.9) ng/L and (9.8±2.1) ng/L, respectively, which both were lower than that of healthy controls ((16.4±2.7) ng/L), and the differences were statistically significant (t=23.107 and 15.853, both P<0.05). The results of IHC indicated that the levels of IL-38 in the intestinal mucosal tissues of UC and CD patients were 0.04±0.01 and 0.03±0.01, respectively, which were both lower than that of healthy controls (0.18±0.02), and the differences were statistically significant (t=48.186 and 69.443, both P<0.05). The levels of NF-κB and p-STAT3 of UC and CD patients were 0.150±0.030, 0.160±0.040 and 0.130±0.030, 0.110±0.010, which were all higher than those of healthy controls (0.020±0.003 and 0.010±0.002), and the differences were statistically significant (tUC=27.273 and 23.078, tCD=23.657 and 62.684; all P<0.05). The number of patients with disease at active phase, CRP level, ESR and disease activity scores of UC and CD patients with low IL-38 expression were all significantly higher than those of patients with high IL-38 expression (χUC2=11.552, tUC=7.118, 8.991 and 7.086; χCD2=5.675, tCD=9.559, 9.358 and 11.268; all P<0.05). The results of Pearson correlation analysis demonstrated that the level of IL-38 in the intestinal mucosal tissue of UC patients was negatively correlated with CRP, ESR and Mayo scores (r=-0.291, -0.672 and -0.639; all P<0.05). And the level of IL-38 in the intestinal mucosal tissue of CD patients was negatively correlated with CRP, ESR and CDAI (r=-0.559, -0.471 and -0.353; all P<0.05). The IHC results showed that the levels of NF-κB and p-STAT3 of IL-38 transgenic IBD mice were lower than those of wild type IBD mice (0.14±0.02 vs. 0.32±0.06, 0.12±0.02 vs. 0.44±0.07), and the differences were statistically significant (t=6.971 and 10.767, both P<0.05). The results of flow cytometry showed that the ratio of CD4+ IL-17+ Th17 cells in the peripheral blood of IL-38 transgenic IBD mice was lower than that of wild type IBD mice (0.030±0.006 vs. 0.280±0.050), and the difference was statistically significant (t=12.160, P<0.05). Conclusions The expression level of IL-38 significantly decreases in the intestinal mucosal tissues of IBD patients, while the level of NF-κB and p-STAT3 significantly increases. IL-38 may inhibit IBD by regulating NF-κB and p-STAT3 signaling pathway to alleviate intestinal immune reaction. Key words: Inflammatory bowel diseases; NF-kappa B; interleukin-38; STAT3

Combination of L-Arginine and L-Norvaline protects against pulmonary fibrosis progression induced by bleomycin in mice

Pulmonary fibrosis (PF) progression may be involved with arginine (Arg) metabolism and immune balance. The present study aimed to explore the effects of L-Arginine (L-Arg) and L-Norvaline (L-Nor) on bleomycin (BLM)-induced PF in mice, meanwhile, and observe dynamic changes of Arg metabolism, immune balance and crosstalk between them in PF progression. Followed intratracheal instillation of BLM or saline, Kunming mice were treated orally with saline, L-Arg, L-Nor and L-Arg + L-Nor three times a day. And the mice were sacrificed on Day 3, 14 and 28 after treatment. Changes of body weight, lung index, lung hydroxyproline and histopathology were analyzed to evaluate the PF degree. Peripheral blood Arg, Citrulline (Cit), Ornithine (Orn) and Proline (Pro), lung NO, NOS and arginase were analyzed to evaluate the Arg metabolism. Peripheral blood Tregs, Th17 and γδT cells were analyzed to evaluate the immune balance. Our data showed that combination of L-Arg and L-Nor dynamically reversed the weight loss, decreased lung index and hydroxyproline, and improved lung histopathological damages induced by BLM. The combination dynamically and significantly rectified Tregs, Th17, γδT and Tregs/Th17 abnormal changes. Meanwhile, these disorders of peripheral blood Arg, Cit, Orn, Pro, Orn/Cit and Pro/Orn, and lung NO, iNOS and TNOS were also improved accordingly. These results demonstrated that combination of L-Arg and L-Nor had inhibitory effects on BLM-induced PF progression, possibly due to their corrective action on immune imbalance, Arg metabolism disorder and crosstalk abnormality in the progression of PF.

Characterization of lymphocyte subsets in peripheral blood cells of children with EV71 infection.

Enterovirus 71 (EV71) is one of the major causative pathogens of hand, foot, and mouth disease (HFMD). Immune cells play a critical role in determining the outcomes of virus infection. We aimed to characterize the lymphocyte subsets and transcriptional levels of T lymphocytes-associated transcription factors in peripheral blood cells of children with EV71 infection. Peripheral blood samples from 32 children with EV71 infection and 32 control subjects were included in this study. The frequencies of T-, B-lymphocytes, and their subsets were determined by flow cytometry. The expression of transcription factors, including T-bet, Gata3, ROR γ t, Foxp3, TCF-1, and BCL-6 in the whole blood cells were evaluated by real-time reverse-transcription quantitative polymerase chain reaction (RT-qPCR). The frequencies of T cells, helper T cells (Th), cytotoxic T cells (Tc), IFN-γ+ Th1, IFN-γ+ Tc1, and regulatory T (Treg) cells were significantly decreased (P<0.01) in children with EV71 infection. As for IL-4+ Th2, IL-4+ Tc2, IL-17+ Th17, IL-17+ Tc17, follicular helper T cells (Tfh), CD3+CD8+IL-21+ T cells, CD19+ B cells, and CD19+IL-10+ B10cells, their frequencies were significantly increased in the EV71 group (P<0.01). The EV71 group had lower mRNA expressions of T-bet, Gata3, and Foxp3 than the control group (P<0.05), whereas the expressions of ROR γ t, TCF-1, and BCL-6 showed no significant difference between two groups. EV71 infection in children caused a decreased frequency of total Th, Tc and Treg cells, and increased percentages of B cell, Th2 and Th17cells in blood.

Research of the correlation between peripheral blood Th17/Treg balance and cerebral infarction

Objective To investigate the correlation between Th17/Treg balance in peripheral blood and cerebral infarction and its clinical value. Methods One hundred and twenty-two patients with cerebral infarction treated in affiliated Dongfeng Hospital of Hubei University of Medicine from February 2013 were randomly selected as the infarction group, and the other 122 healthy subjects for physical examination in Dongfeng Hospital at the same period were selected as the control group.The two groups were tested for peripheral blood Th17/Treg cells, and the content of interleukin-17 was detected and the correlation was analyzed. Results The percentage of Th17 cells and the levels of IL-17 in infarction group were (1.45±0.24)% and (58.20±22.19) ng/L, respectively, which were higher than those in the control group ((0.84±0.15)%, (21.00±11.93) ng/L)(t=10.648, 11.285, P<0.05). The proportion of Treg cells in the infarction group was (4.89±0.81)%, lower than that of the control group ((11.38±1.89)%) (t=11.298, P<0.05). Spearman rank correlation analysis showed that the proportion of Treg cells in patients with cerebral infarction were negatively correlated with Th17 cell ratio, IL-17 content, neurological deficit score and infarct volume (r=-0.671, -0.385, -0.414, -0.533, P<0.05). Multivariate logistic analysis showed that the percentage of Th17, IL-17 content and infarct volume were the main influencing factors of Treg cells (OR(95%CI)=2.216(1.120-4.283), 2.062(1.109-3.298), 1.022(1.008-4.192), P<0.05). Conclusion Peripheral blood Th17 cells and Treg cells have the proinflammatory and anti-inflammatory effects by secreting cytokines in patients with acute cerebral infarction.Th17/Treg balance in peripheral blood of patients with acute cerebral infarction is involved in the process of brain tissue damage after cerebral infarction. Key words: Cerebral infarction; Th17 cells; Treg cells; Interleukin-17

Impact of HIV-ART on the restoration of Th17 and Treg cells in blood and female genital mucosa

The aim of this study was to evaluate the effectiveness of antiretroviral treatment (ART) on the proportion and functions of Th17 and Treg cells in peripheral blood and female genital tract (FGT) respectively. To this aim, samples from 41 HIV-neg, 33 HIV+ ART-naïve and 32 HIV+ ART+ subjects were obtained. In peripheral blood, altered Th17 and Th17/Treg proportions were normalized in HIV+ ART+, but certain abnormal Treg and activated T-cell proportions were still observed. In FGT, abnormal patterns of secretion for Th17-related cytokines were observed in cervical mononuclear cells (CMCs) from HIV+ women, even in those from HIV+ ART+, compared to the HIV-neg group. Moreover, these altered patterns of secretion were associated with diminished levels of CXCL5 and CXCL1 chemokines and with an immunoregulatory skew in the CCL17/CCL20 ratio in ectocervix samples of these women. Finally, ART did not restore proportions of Th17-precursor cells with gut-homing potential in PBMCs, and positive correlations between these cells and the levels of IL-17F and IL-21 production by CMCs may suggest that a better homing of these cells to the intestine could also imply a better restoration of these cells in the female genital tract. These results indicate that antiretroviral treatment did not restore Th17-related immune functions completely at the female mucosal level.

Role of Th9, Th17, Treg Cells levels and IL-9, IL-17 and TGF-β Expression in Peripheral Blood of Patients with ITP in Pathogenesis of ITP

To detect the levels of Treg, Th17, Th9 cells and expression of transforming growth factor-β (TGF-β), interleukin-17 (IL-17) and interleukin-9 (IL-9) in peripheral blood of patients with immune thrombocytopenia (ITP) and to explore its role in the pathogenesis of ITP. Fifty-four patients with ITP (ITP group) and 40 healthy volunteers (control group) were selected in our hospital. The of Treg, Th17 and Th9 cells in peripheral blood of 2 groups were measured by flow cytometry, and the expression of cytokines, such as TGF-β, IL-17 and IL-9 in the peripheral blood of 2 groups were detected by enzyme linked immunosorbent assay (ELISA). The level of Treg cells in the peripheral blood of the ITP group was significantly decreased in comparison with the control group, while the levels of Th17 and Th9 cells significantly increased in comparison with the control group (all P<0.01). The expression of cytokine such as TGF-β in the peripheral blood of the case group significantly decreased in comparison with the control group, while the expression levels of IL-17 and IL-9 significantly increased in comparison with the control group (P<0.01). The results of Pearson correlation analysis showed that there was a positive correlation between the level of Treg cells and platelet count (PLT) in peripheral blood of the ITP group (r=0.35, P<0.05), and there were negative correlation between the level rate of Th17, Th9 cells and Plt count (r=-0.37, -0.43, P<0.05); there was a positive correlation between the expression of the TGF-β in the ITP group and Plt count (r=0.46, P<0.05), while the expression of IL-17 and IL-9 showed negative correlation with PLT (r=-0.48, -0.54, P<0.05). The percentage of Treg, Th17 and Th9 cells in the peripheral blood of patients with ITP is abnormal, and the expression of TGF-β, IL-17 and IL-9 also is abnormal, which may play an important role in the pathogenesis of ITP.

Pregnancy Epigenetic Signature in T Helper 17 and T Regulatory Cells in Multiple Sclerosis.

Increasing evidence supports the anti-inflammatory role of estrogens in Multiple Sclerosis (MS), originating from the observation of reduction in relapse rates among women with MS during pregnancy, but the molecular mechanisms are still not completely understood. Using an integrative data analysis, we identified T helper (Th) 17 and T regulatory (Treg) cell-type-specific regulatory regions (CSR) regulated by estrogen receptor alpha (ERα). These CSRs were validated in polarized Th17 from healthy donors (HD) and in peripheral blood mononuclear cells, Th17 and Treg cells from relapsing remitting (RR) MS patients and HD during pregnancy. 17β-estradiol induces active histone marks enrichment at Forkhead Box P3 (FOXP3)-CSRs and repressive histone marks enrichment at RAR related orphan receptor C (RORC)-CSRs in polarized Th17 cells. A disease-associated epigenetic profile was found in RRMS patients during pregnancy, suggesting a FOXP3 positive regulation and a RORC negative regulation in the third trimester of pregnancy. Altogether, these data indicate that estrogens act as immunomodulatory factors on the epigenomes of CD4+ T cells in RRMS; the identified CSRs may represent potential biomarkers for monitoring disease progression or new potential therapeutic targets.

Comparison of Peripheral Blood Th17 Cells and Associated Cytokines in Fingolimod-Receiving and Untreated Multiple Sclerosis Patients

Comparison of Peripheral Blood Th17 Cells and Associated Cytokines in Fingolimod-Receiving and Untreated Multiple Sclerosis Patients

Effects of Early Enteral Nutrition on Immune Function and Prognosis of Patients With Sepsis on Mechanical Ventilation.

To explore the therapeutic effects of early enteral nutrition (EEN) on patients with sepsis on mechanical ventilation. Patients with sepsis on mechanical ventilation in the medical intensive care unit (ICU) from January 2013 to March 2016 were treated with enteral nutrition. Patients treated within 48 hours of initiation of mechanical ventilation were assigned to the EEN group, and the rest were assigned to the delayed enteral nutrition (DEN) group. Peripheral blood Th17 cells and Treg cells, endotoxin (ET) level, 28-day mortality, duration of mechanical ventilation, lengths of ICU stay and hospital stay, and incidence of ICU-acquired weakness (ICU-AW) were analyzed between the 2 groups. The proportion of Th17 cells and ET levels in the EEN group were significantly lower than those in the DEN group, whereas the proportion of Treg cells in the EEN group was remarkably higher than that in the DEN group (P < .05). The duration of mechanical ventilation, lengths of ICU stay and hospital stay, and incidence of ICU-AW were higher in the DEN group than in the EEN group (P < .05), but there was no significant difference in the 28-day mortality between the 2 groups. Patients with sepsis mainly present with an increased proportion of Th17 cells in the early stage, manifesting as enhanced immune response. Early enteral nutrition can inhibit the excessive immune response, shorten the duration of mechanical ventilation, lengths of ICU stay and hospital stay, and reduce the incidence of ICU-AW, but it has no obvious effect on 28-day mortality.

IL-17A contributes to HSV1 infection-induced acute lung injury in a mouse model of pulmonary fibrosis.

BackgroundPatients with idiopathic pulmonary fibrosis (IPF) often experience acute exacerbation (AE) after an episode of common cold.AimsTo establish a mouse model of virus infection‐induced AE‐IPF and investigate the mechanism underlying the AE‐IPF.MethodsHerpes simplex virus 1 (HSV1) was inoculated intranasally to wild‐type (WT) and IL‐17A gene knockout (IL‐17A‐/‐) mice 21 days after intratracheal administration of bleomycin (BLM).Results HSV1 infection caused acute exacerbation in mice with BLM‐induced fibrosis. Compared with the BLM+Saline mice, the mice with BLM+HSV1 showed significantly higher acute lung injury (ALI) score (P < 0.0001), lower survival rate (100% vs 21.4%, P < 0.0001), poorer lung function and higher inflammatory response representing by increased total inflammatory cells in bronchoalveolar lavage fluid (BALF) (P = 0.0323), increased proportion of Th17 cells in peripheral blood (P = 0.0004) and higher inflammatory factors in BALF. In addition, HSV1 infection increased the expression of endoplasmic reticulum stress (ERS)‐related proteins in mice with BLM‐induced fibrosis. The inhibition of ERS by tauroursodeoxycholic acid (TUDCA, an ERS inhibitor) significantly reduced the IL‐17A levels in BALF (P = 0.0140) and TH17 cells in the peripheral blood (P = 0.0084) of mice with BLM+HSV1, suggesting that suppression of ERS may reduce TH17 response in mice with AE‐IPF. Compared with WT mice with BLM+HSV1, IL‐17A‐/‐ mice with BLM+HSV1 had lower ALI score (P = 0.0119), higher survival rate (78.6% vs 21.4%, P = 0.004), improved lung function, and milder inflammatory response.Conclusions HSV1 infection in addition to BLM‐induced IPF can successfully establish AE‐IPF in mice. IL‐17A and ERS promote lung inflammation in AE‐IPF development.

Role of Th17 Cells, Interleukin-17 and Matrix Metalloproteinase-13 in Pathogenesis of Henoch-Schonlein Purpura

To explore the immunopathogenesis of Henoch-Schonlein purpara (HSP) by detecting the levels of Th17 cells, IL-17 and matrix metallo proteinase-13 (MMP-13) in peripheral blood and the expression of IL-17 in skin lesions at acate phase of HSP. Th17 cell ratio in peripheral blood of HSP group and healthy control group was defected by flow cytometry, the plasma levels of IL-17 and MMP-13 in HSP group and healthy control group were defected by ELISA, and expression level of IL-17 in skin lesion of HSP group and skin tissue of healthy control group was deternined by: immunohistochemistry method. the ratio of Th17 cells in periphral blood of HSP group (1.21±0.59%) was very signif cantly higher than that in peripheral blood of control group (0.71±0.26%) ( t=4.907, P<0.01). The plasma levels of IL-17 and MMP-13 at acute phase of HSP were very significantly higher than those in control group (64.58±36.21) pg/ml vs (26.16±14.90) pg/ml and (17.57±5.40) pg/ml vs (11.53±4.40) pg/ml respectively (t=6.183, P<0.01 and t=5.022, P<0.01). The integrool optical density of IL-17 in skin lesin tissue of HSP group (7.26±2.34) was higher than that in control group (4.61±1.82) ( t=2.877, P<0.01). Th17 cells, IL-17 and MMP-13 may be involved in the immunological pathogenesis of HSP.

Peripheral Th17 cells expressing β7 intestinal homing receptor in recent and chronic HIV infections.

The objective of this study was to conduct an analysis of peripheral blood Th17 cells with the ability to home to gut mucosa (CD4+ Th17+ β7+ ) during recent or chronic human immunodeficiency virus (HIV) infections. The relationship between HIV load and systemic inflammation markers was studied. Twenty-five patients with recent (n=10) or chronic (n=15) untreated HIV infections; 30 treated HIV-infected patients with undetectable HIV load at the time of inclusion and 30 healthy controls were included. Bacterial translocation markers (16S rDNA), soluble CD14 (sCD14) and interleukin (IL)-6 monocyte activation parameters, CD4/CD8 ratio and T helper type 17 (Th17) subpopulations [CD4+ Th17+ expressing the IL-23 receptor (IL-23R) or β7] were analysed at baseline and after 6 and 12months of anti-retroviral therapy (ART). 16S rDNA was detected in all patients. Significantly increased serum levels of sCD14 and IL-6 and a decreased CD4/CD8 ratio were observed in patients. Similar percentages of CD4+ IL-23R+ and CD4+ Th17+ β7+ cells were observed in healthy controls and patients at baseline. After 12months of therapy, patients with a recent HIV infection showed significant increases of CD4+ IL-23R+ and CD4+ Th17+ β7+ cell percentages and a decrease in IL-6 levels, although 16S rDNA continued to be detectable in all patients. No significant differences were observed in Th17 subpopulations in patients with chronic HIV infection after therapy. Early initiation of ART helps to increase the number of Th17 cells with the ability to home to the intestinal mucosa and to partially restore gut mucosal homeostasis. These results provide a rationale for initiating ART during the acute phase of HIV infection.

Lung-Resident Mesenchymal Stem Cells Promote Repair of LPS-Induced Acute Lung Injury via Regulating the Balance of Regulatory T cells and Th17 cells.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are associated with high morbidity and mortality. Mesenchymal stem cells (MSCs) have been shown to improve ALI, and the imbalance of regulatory T cells (Tregs) and Th17 cells is associated with mortality in ALI/ARDS patients. However, whether administration of lung-resident MSC (LRMSC) improves lung injury and regulates the balance of Tregs and Th17 cells remains unknown. An ALI animal model was induced by LPS, and PBS or LRMSC were administered via tail vein after 4h. LRMSC were subsequently detected in the lungs by a live imaging system (Berthold LB983, Germany). Lung morphology; lung wet-to-dry weight ratio; and total protein concentration, inflammatory cells, and cytokines in bronchoalveolar lavage fluid (BALF) and plasma were determined. The percentage of Tregs in lung and spleen, and of Th17 cells in lung and blood, were also evaluated. The results showed that LRMSC not only attenuated histopathological damage but also mediated the downregulation of lung wet-to-dry weight ratio and the reduction of total protein concentration and inflammatory cells in BALF. LRMSC also decreased inflammatory cytokines in both BALF and plasma and increased KGF-2 and surfactant protein C (SPC) expression in the lung. Flow cytometry revealed the upregulation of Tregs and the downregulation of Th17 cells, and the increase in the ratio of Tregs and Th17 cells. The live imaging system showed that LRMSC migrated to and were retained in the injured area. In conclusion, the results indicated that administration of LRMSC attenuates LPS-induced ALI via upregulating the balance of Tregs and Th17 cells.

Interleukin-17 inhibits the growth of oral squamous cell carcinoma by promoting the differentiation of T helper 17 cells

Background: The infiltration of T helper 17 (Th17) cells has been reported in multiple solid tumors. Recent studies implicated that Th17 cells played an inhibitory role in the tumor growth. However, if Th17 cells infiltrated and played a role in oral squamous cell carcinoma (OSCC) remained unknown. Methods: Fluorescence activated cell sorting (FACS) was performed to confirm the increase of Th17 cells in the peripheral blood and the infiltration in tumor samples from OSCC patients. To mimicking the effect of Th17 cells on OSCC growth, an OSCC line stably expressing human interleukin-17A (IL-17A) was established. Human-PBL-SCID-beige mice were employed as the hosts for the implantation of OSCC/IL-17A cells. Quantitative PCR, ELISA and Western blotting were performed to analyze Th17 differentiation and function in the implanted tumors. Results: The percentage of Th17 cells increased significantly in both the peripheral blood and the tumor tissues from OSCC patients. Expression of IL-17A promoted the differentiation of Th17 cells and suppressed the growth of implanted OSCC tumors. The transcription as well as the protein levels of IL-6 and granzyme B in the implanted OSCC cells was remarkably elevated by IL-17A expression. Conclusions: The increased infiltration of Th17 cells was suggested to result in a therapeutic effect on OSCC as Th17 cells did in other tumors. IL-17 could promote the differentiation of Th17 cells by increasing IL-6 expression. The inhibitory effect of Th17 cells on OSCC growth was associated with their increased cytotoxic activity.

Spontaneous Diabetic FATZO Mice with Intact Leptin Signaling Develop Peripheral Immune Cell Responses Similar to T2D and Obese Patients

Introduction: Type 2 diabetes (T2D) and obesity are strongly associated with peripheral immune cell responses. Subsets of T cells in the circulation are correlated with the severity of the diseases, which may serve as novel targets for antidiabetic or obese treatment. Higher numbers of blood Th1, Th17 and Th22 cells of T helper cells were found in disease patients, while Th2 and Treg cells were in reverse correlation of the severity of the disease. FATZO mouse is a new generation of T2D mouse model with intact leptin signaling. The model is featured with slow onset of diabetes and obesity. Here, we aim to characterize the peripheral immune cell responses in FATZO mice for the use of preclinical drug development targeting diabetes or obesity related inflammation. Methods: Freshly collected blood from 12 FATZO mice at age 6, 12, 22 and 34 weeks were analyzed for the presence of monocytes, B, T, NK, NK-T, CD4+, CD8+ T cells and subsets of T helper cells (Th1, Th2, Th17, Th22, Treg) using FACS. Serum inflammatory cytokines such as IL1, IL2, IL6, IL17, TNFα and IFNγ were also measured. Results: FATZO mice developed obesity and hyperglycemia from age of 12 weeks. Concomitantly, higher numbers of leukocytes were noticed with an increase and decrease in frequency of B and NK cells respectively. Within T cells, numbers of CD4+ T cells decreased by 20%, while CD8+ T cells increased by 25%. When subsets of T helper cells were assessed, percentages of Th1 and Th17 cells significantly elevated during the development of diabetes and obesity, while frequencies of Th2 and Treg cells declined. Serum inflammatory cytokine measurements showed a tendency in higher levels of serum IL1a when the animals became diabetic. Conclusion: FATZO mice develop diabetes and obesity with changes in blood immune cell responses that closely mimic disease patients. The results suggest that FATZO might serve as an ideal model for preclinical research for the diabetes and obesity related inflammation. Disclosure G. Sun: None. W. Ye: Employee; Spouse/Partner; Crown Bioscience Inc.. X. An: None. Y. Xiao: None. Y. Wang: None.

Higher levels of serum interleukin-35 are associated with the severity of pulmonary fibrosis and Th2 responses in patients with systemic sclerosis.

The aim of the study is to investigate the levels of serum interleukin-35 (IL-35) before and after treatment and its relationship with clinical parameters as well as the frequency of different subsets of CD4+ T cells in patients with systemic sclerosis (SSc) to explore the role of IL-35 in the pathogenesis of SSc. The levels of serum IL-35, interferon-γ (IFN-γ), IL-4, IL-17A, and IL-10 in 49 patients with SSc and 50 age- and gender-matched healthy subjects were measured by enzyme-linked immunosorbent assay (ELISA). The percentages of peripheral blood Th1, Th2, Th17 cells and Tregs in 49 SSc patients and 20 healthy controls were characterized by flow cytometry. The relationship between the levels of serum IL-35 and the percentages of different subsets of CD4+ T cells, disease duration, the values of forced vital capacity (FVC), modified Rodnan skin scores (MRSS) or high-resolution computed tomography (HRCT) scores was analyzed in patients with SSc. The levels of serum IL-35 in SSc patients were significantly higher than that of healthy controls (P < 0.001), but significantly reduced after treatment for 3months (P = 0.001). The levels of serum IL-35 were positively correlated with the HRCT scores in SSc patients (P = 0.014) and with disease duration in patients with diffuse cutaneous SSc (P = 0.03), but negatively correlated with the FVC values in SSc patients (P = 0.034). In comparison with that in the healthy controls, significantly decreased percentages of Th1, but increased Th2 and Th17 cells were detected in patients, leading to reduced ratios of Th1/Th2 and increased ratios of Th17/Tregs (P < 0.001). The levels of serum IL-35 were positively correlated with IL-4 (P = 0.036) and tended to be positively associated with the frequency of Tregs in SSc patients (P = 0.054). Higher levels of serum IL-35 may be associated with the development of SSc and severity of pulmonary fibrosis in SSc patients.

Th17 immune responses in Brazilian dyslipidemic patients with atherosclerosis

Lymphocytes, macrophages, and immunological mediators are critical in atherogenesis, but Th17 cells are still little studied. We investigated blood lymphocyte profiles, cytometric changes in blood Th17 cells, and production of Th17-associated cytokines by peripheral blood mononuclear cells (PBMCs) stimulated with phytohemagglutinin (PHA) in dyslipidemic patients. Forty-nine Brazilian dyslipidemic patients, including 14 with negative coronary angiograms (CAs) and 35 with positive CAs (atherosclerosis), were classified into groups G1 and G2, respectively, and 27 healthy individuals without dyslipidemia were included in the control group (HI). In all participants, lipid profiles, CVD risk factors, atherogenic risk indexes, cytometry of blood T lymphocytes (CD4+ T, CD8+ T, and Th17 cells), and Th17 cytokine production were determined. Production of interleukin (IL)-17A, IL-17F, IL-21, and IL-22 was measured in culture supernatants of PHA-stimulated PBMCs. Patients with atherosclerosis exhibited low levels of high-density lipoprotein cholesterol and apolipoprotein A and showed increased atherogenic risk indexes and a higher frequency of prior acute myocardial infarction (AMI). CD8+ T-cell counts were lower in patients with previous AMI and higher in dyslipidemic obese individuals. Th17 cell frequency was increased in dyslipidemic women with atherosclerosis. PBMCs from patients with atherosclerosis produced less IL-17A and IL-21, but more IL-22. IL-17F production was unaltered. In summary, cytometric changes in CD8+ T and Th17 lymphocytes and altered production of Th17-associated cytokines were observed in these individuals and were related to endocrine factors, obesity, and AMI. The practical implications of these findings need to be investigated in future studies.

Relationship Between Dynamic Changes in Expression of IL-17/IL-23 in Lacrimal Gland and Ocular Surface Lesions in Ovariectomized Mice.

An ovariectomized mouse model was constructed to observe the dynamic effects of hormone changes on the expression of interleukin (IL)-17A and IL-23 in the lacrimal glands. The ovariectomized mouse model was constructed by bilateral ovary removal. The concentrations of serum estradiol and testosterone in mouse cardiac blood were detected by radioimmunoassay. Mice in both groups underwent the phenol red cotton thread test and corneal fluorescein staining to assess the ocular surface, whereas Th17 cells in blood and spleen were detected by flow cytometry. IL-17A and IL-23 expression in the lacrimal glands was detected by immunohistochemistry, enzyme-linked immunosorbent assay, and real-time fluorescence quantitative polymerase chain reaction. Serum estradiol and testosterone levels were significantly lower in the ovariectomized group compared with those in the control group. There was lymphocytic infiltration in the lacrimal gland of the ovariectomized group mice. At 6 months after the surgery, aqueous tear production was significantly lower, and statistically significant corneal fluorescein staining was found in the ovariectomized group, compared with that in the control group. In the ovariectomized group, IL-17A and the IL-23 expression in the lacrimal glands and the Th17 expression in the blood and spleen were significantly higher than in the control group. The hormone levels are significantly reduced and lymphocytic infiltration in the lacrimal gland in ovariectomized mice, whereas the frequency of Th17 cells in the blood and spleen and IL-17A and IL-23 expression in the lacrimal glands are increased, leading to reduced tear production and positive fluorescein staining in the cornea.

Activated Α7nachr Improves Postoperative Cognitive Dysfunction and Intestinal Injury Induced by Cardiopulmonary Bypass in Rats: Inhibition of the Proinflammatory Response Through the Th17 Immune Response

Backgrund/Aims: To investigate the effects of activated α7 nicotinic acetylcholine receptor (α7nAChR) on postoperative cognitive dysfunction (POCD) and intestinal injury induced by cardiopulmonary bypass (CPB) and its relationship with the Th17 response in order to provide a theoretical basis for organ protection and targeted drug therapy during the perioperative period. Methods: Sprague-Dawley rat models of CPB were established. Rat intestinal and brain injuries were observed after CPB using hematoxylin and eosin staining. Cell apoptosis was determined using terminal deoxynucleotidyl transferase dUTP nick end labeling. Inflammatory factors and markers of brain injury in rat serum were measured using enzyme-linked immunosorbent assay. The expression levels of Bcl-2, Bax, caspase-3, ZO-1, occludin, AQP4, RORγT, and α7nAchR were examined using western blotting. Transcription factor RORγT expression was determined using real-time fluorescent quantitative polymerase chain reaction. Th17 cells in the peripheral blood and spleen were determined using flow cytometry. α7nAchR knockout rats were established. The Th17 response in the peripheral blood and spleen of α7nAchR knockout rats was further verified using flow cytometry. Results: CPB can induce POCD and intestinal injury in rats. α7nAchR activation markedly reduced intestinal injury, POCD, neuronal apoptosis, proinflammatory factor expression, and number of CD4⁺IL-17⁺ cells. α7nAchR knockout significantly increased serum D-lactic acid, FABP2, S-100β, NSE, TNF-α, IL-6, and IL-17 secretion. The number of CD4⁺IL-17⁺ cells was also significantly increased. Conclusion: α7nAchR activation markedly ameliorates the intestinal injury and POCD induced by CPB. Inhibition of the Th17 immune response can reduce the proinflammatory response, which could provide a new method for clinical perioperative organ protection and targeted drug therapy.

Change of Peripheral Blood Treg/Thl7 in Cognitive Impairment with Chronic Renal Failure Patients

Background/Aims: To investigate the changes in peripheral blood Treg/Th17 cell balance and its significance in patients with chronic renal failure (CRF) and cognitive impairment. Methods: A total of 71 patients with CRF were enrolled as a study group. The patients were divided into a cognitive impairment group and a normal cognitive function group according to the Mini-Mental State Examination (MMSE). Peripheral blood Treg and Th17 cells were analyzed by flow cytometry and their relevant cytokines (IL-17, IL-10 and TGF-β) and other biochemical indicators, including C-reactive protein (CRP) and IL-6, were determined by ELISA. Results: Thepatients with both CRF and cognitive impairment were older than the cognitive normal groups. Peripheral blood Treg cells by Flow cytometry (the CRF cognitive impairment group 5.57±1.3%, CRF group with normal cognitive function 7.5 ± 0.9% and normal control group 9.7 ± 1.7%,P<0.05) and its related cytokines (IL-10 and TGF-β) by ELISA detection were lower in the group with cognitive impairment than in the group without cognitive impairment ( IL-10, 7.4±4.2 pg/mL, 13.8±3.9 pg/mL, 18.3±3.2 pg/mL; TGF-β 335.6±175.3 pg/mL, 512.7 ± 114.6 pg/mL, 953.8±373.4 pg/mL P < 0.05, respectively).However, Th17 cell numbers (the CRF cognitive impairment group 3.3 ± 0.7%, CRF group with normal cognitive function2.2 ± 0.5% and normal control group 1.5 ± 0.3%),and cytokine levels (IL-17, IL-6 and CRP) were higher in the group with cognitive impairment IL-6 (21.3 ± 5.1 pg/mL), IL-17 (18.5 ± 4.2 pg/mL) and CRP (20.3 ± 5.9 mg/L) in the CRF group with cognitive impairment when compared with the CRF group and normal cognitive function (12.2 ± 4.5 pg/mL, 12.1 ± 3.7 pg/mL and 13.5 ± 4.6 mg/L, respectively) or the normal control group (9.2 ± 5.8 pg/mL, 7.4 ± 2.6 pg/mL and 3.2 ± 1.3 mg/L, respectively, P<0.05). The frequencies of Treg in patients with CRF were positively correlated with the MMSE scores ((r = 0.518, P < 0.05), but the Th17 numbers were negatively correlated (r = -0.435, P < 0.05). Conclusion: An imbalance of peripheral blood Treg/Th17 cells is associated with cognitive impairment in patients with CRF.