Blood Th17 Research Articles

Bone marrow IRF4 level in multiple myeloma: an indicator of peripheral blood Th17 and disease.

Interferon regulator factor 4 (IRF4) is characterized to be a member of interferon regulatory family, which is predominantly expressed in bone marrow plasma cells of patients with multiple myeloma (MM). Recent studies indicated IRF4 is critical for T-help cells (Th17) differentiation and interleukin-17 (IL-17) secretion. Here, a total of 58 MM patients were enrolled in this study, the proportions of Th17 cells and T regulatory (Treg) cells in peripheral blood mononuclear cells (PBMCs) were determined by flow cytometric analysis. Immunohistochemistry was employed to detect the IRF4 expression in bone marrow. Herein, we observed a significant increase of IRF4 in bone marrow accompanied with a notable up-regulation of Th17 cells in PBMC within MM patients compared with healthy donors. Furthermore, the proportions of Th17 cells and serum IL-17 levels were higher in patients with stage III than stage I & II MM patients, and those parameters were positively correlated with the expression of IRF4 in these cases. These results for the first time indicate that a crosstalk between IRF4 and Th17 cells is associated with MM prognosis, and IRF4 may be served an important target for MM immunotherapy.

Crossover Subsets of CD4+ T Lymphocytes in the Intestinal Lamina Propria ofPatients with Crohn's Disease and Ulcerative Colitis.

Hovhannisyan et al. first showed evidence of plasticity between Treg and Th17 in the inflamed intestine of Crohn's disease (CD) patients. Our previous report suggests that the inflammatory cytokine milieu generates IL-17+ Foxp3+ CD4+ T lymphocytes which is a crossover population converting Treg subset to Th17 in the peripheral blood of IBD patients. This is considered as an evidence of Treg/Th17 plasticity. The aim of this study was to characterize a variety of helper T cell crossover population, not limited to IL-17+ Foxp3+ CD4+ T lymphocytes, in the lamina propria (LP) of IBD patients. Fresh colonoscopic biopsies were obtained from patients with CD (n=50) and ulcerative colitis (UC, n=32) and from healthy controls (HC, n=25). LP mononuclear cells were assessed for intracellular cytokines and transcription factors such as IFNγ, IL-13, IL-17, IL-22, T-bet, Gata-3, RORγt, and Foxp3 using multicolor flow cytometry to detect subsets of LP CD4+ T lymphocytes. Patients with IBD demonstrated increased crossover populations in IL-17+ Foxp3+, T-bet+ Foxp3+, Gata3+ Foxp3+, RORγt+ Foxp3+ populations compared to HC. There was an inverse correlation of Harvey-Bradshaw index with Gata3+ Foxp3+ population in CD patients, while IL-13+ Foxp3+ population was directly correlated with Mayo clinical scores in UC patients. Furthermore, total IL-22 expressing cells as well as Th22 and IL-22+ Th1 populations were decreased in UC compared to CD and HC. IBD patients exhibit the increased crossover populations in LP Treg cells toward Th2 and Th17 compared to HC. The prevalence of Treg/Th2 crossover populations is associated with clinical disease score of IBD.

Immunosuppressive treatment using tacrolimus promotes pregnancy outcome in infertile women with repeated implantation failures.

We aim to investigate whether the peripheral blood T helper (Th) 1 cell level could predict pregnancy outcome in patients who have experienced repeated implantation failure (RIF, three or more) after ART cycles. This is a prospective cohort study of total 124 women with RIF who showed elevated Th1/Th2 (CD4+ IFN-γ+ /CD4+ IL-4+ ) cell ratios (≥10.3) and received tacrolimus at Sugiyama Clinic between November 2011 and July 2016. Patients were divided into three groups as per Th1 cell levels: Th1 level of <22.8 as Low; 22.8 to <28.8 as Middle, and 28.8 or greater as High group. The study patients received daily dose of tacrolimus 1-3mg based on initial Th1/Th2 cell ratio. The clinical pregnancy rates of Low, Middle, and High groups were 48.8%, 43.9%, and 33.3%, respectively (P=NS), with tacrolimus treatment. The ongoing pregnancy/delivery rate of Low group (46.3%) was significantly higher than that of High group (21.4%, P<.05). Middle group (34.3%) had higher success rate than High group, albeit without statistical significant. We confirm our previous report that Th1/Th2 ratio can predict ART outcomes in patients with RIF and immunosuppressant treatment with tacrolimus, and peripheral blood Th1 cell levels were negatively correlated with pregnancy outcome.

073 Th17 and Th22 polarization in ichthyosis blood correlates with disease severity and highlights its systemic nature

073 Th17 and Th22 polarization in ichthyosis blood correlates with disease severity and highlights its systemic nature

Expression of T helper 17 cells and interleukin 17 in lupus nephritis patients

BackgroundModern data suggest that interleukin 17 (Il-17) and Il-17 producing cells play an important role in the pathogenesis of lupus nephritis (LN). It was reported that T helper 17 migrate to the kidney and contribute to inflammatory processes. Aim of the workTo measure the frequencies of peripheral blood T helper-17 (Th17) cells and IL-17 concentration in urine in systemic lupus erythematosus (SLE) patients and to study their association with disease activity and renal involvement. Patients and methodsThis work included 45 patients; 36 females and 9 males with 20 matching controls. Patients were subjected to clinical and laboratory assessments with emphasis on renal affection. The SLE disease activity index was also calculated. Expression of Th-17 cells in peripheral blood was measured by flow cytometry via the combined expression of CD3, IL-23R, intracellular IL-22, IL-17 and urinary IL-17 was measured by ELISA. ResultsThe patients’ mean age was 26.5±7.6years and disease duration was 6.03±4.3years. Th17 expression and urinary IL-17 were significantly higher in patients than controls (p=0.04 and p<0.001 respectively). Th17 cell frequencies and IL-17 levels significantly correlated with renal biopsy classification for LN (p=0.02 and p=0.04 respectively). Th17 cell expression and IL-17 levels were comparable in SLE patients with and without LN. Th17 cell frequencies significantly correlated with serum creatinine, SLEDAI and inversely with C3 (p=0.003), while IL-17 significantly correlated with CD3 cells, proteinuria and erythrocyte sedimentation rate. ConclusionPeripheral blood Th17 cell frequencies and IL-17 in urine are highly linked to LN in SLE and are promising markers of disease activity in LN. Both are valuable targets for future therapeutic applications.

Corrigendum to "Imbalance of peripheral blood Th17 and Treg responses in children with refractory Mycoplasma pneumoniae pneumonia" [J Infect Chemother 22 (2016) 162-166

Corrigendum to "Imbalance of peripheral blood Th17 and Treg responses in children with refractory Mycoplasma pneumoniae pneumonia" [J Infect Chemother 22 (2016) 162-166

AGE-DEPENDENT CHANGES OF T-REGULATORY AND Th17 SUBSET LEVELS IN PERIPHERAL BLOOD FROM HEALTHY HUMANS

Age-dependent development of Th17 and Treg lymphocyte subsets in healthy humans is studied insufficiently. The present study aimed to investigate quantitative characteristics of Th17 and Treg subsets in peripheral blood of healthy subjects for various age groups. 352 healthy humans (168 female and 184 male), one month to 85 years old, were subject to examination, including 79 infants in their first year of life; 34 children at aged 1 year to 2 years 11 months; 24, at 3 to 4 years 11 months; 28, at the age of 5 to 6 years 11 months; 25 children aged 7-8 years 11 month; 36 children aged 9 to 11 years 11 months; 39 adolescents aged 12 to 14 years 11 months; 26 adolescents aged 15 to 18 years; 25 young adults aged 20 to 35 years; 11 adults at 36 to 49 years old; 16 adults aged 59-70 years, and 9 elderly people over 70 years old. The study was performed with capillary blood in children under 2 years, and venous blood taken in elder persons. The basic and ‘minor’ subsets of peripheral blood lymphocytes were evaluated by flow cytometry using four-color staining of whole blood and following erythrocyte lysis. We used the following surface markers: CD3, CD4, CD8, CD25, CD127, CD161, CD45R0 for lymphocyte subsets detection. It has been shown that Treg percentage (a ratio of CD4 + CD25 hi CD127 low/neg in the CD3 + CD4 + gate) did not depend on age of the people under study, and can be approximated by a linear function. The absolute number of Tregs in childhood is progressively decreased and, after 10 years old, it reaches plateau values. This age-dependent relationship may be approximated by a logarithmic function. Evaluation of Th17 subset levels demonstrated a strong relative and absolute age-dependent growth of this cell subpopulation. Percentage and absolute numbers of Th17 lymphocyte (share of CD4 + CD161 + CD45R0 + in the CD3 + CD4 + gate), can be approximated by a square function. The age of 10-12 years seems to be critical to the immune system formation. We suppose the process of the immune system development to be completed at this age, and maturation of the immune cell populations is then observed. A decrease in both relative and absolute numbers of Treg and Th17 lymphocyte subsets was found in elderly persons (> 70 years old). Our data on peripheral blood Tregs and Th17 subsets, with respect to their percentage and absolute numbers in healthy humans, may be used as age-related reference values.

Peripheral blood T-helper cells and eosinophil populations in patients with atopic and nonatopic chronic rhinosinusitis.

Analysis of recent research indicated that T-helper cells may play an important role in the pathogenesis of chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) and CRS without nasal polyps (CRSsNP). The purpose of this study was to investigate the peripheral blood Th1 and Th2 cells and eosinophil population in patients with CRS. Peripheral blood samples were obtained from nine nonatopic controls, 37 patients with CRSsNP, and 66 patients with CRSwNP. The samples were then analyzed by flow cytometry analysis (Th1 cell [CD4+, interleukin 4-, interferon γ+]; and Th2 cell [CD4+, interleukin 4+, interferon γ-]). The patients were stratified into four groups based on their allergic status by using skin-prick test results and immunoglobulin E level measurements as the following: (1) nonatopic CRSsNP, (2) nonatopic CRSwNP, (3) atopic CRSsNP, and (4) atopic CRSwNP. Eosinophil counts were also compared. The severity of nasal diseases in these patients was assessed via the Lund-Mackay score. No significant differences in peripheral blood Th1 and Th2 cells were found among all the atopic, nonatopic CRS groups, and the nonatopic control groups. Peripheral blood eosinophil levels in atopic CRSwNP were significantly elevated compared with the nonatopic controls (p < 0.05), but no significant difference was found among all atopic and nonatopic CRS groups. Analysis of our data demonstrated that a proportion of systemic Th1- and Th2-skewed lymphocytes in all CRS groups were similar to that in healthy subjects, irrespective of atopic status. The patients with CRSwNP and with atopy but not the patients with CRSsNP and with atopy demonstrated systemic eosinophilic inflammation. Further studies are needed to investigate underlying pathophysiologic mechanism or endotypes.

Abstract B90: PFK-158 is a first-in-human inhibitor of PFKFB3 that selectively suppresses glucose metabolism of cancer cells and inhibits the immunosuppressive Th17 cells and MDSCs in advanced cancer patients

Abstract Background: 6-phosphofructo-2-kinase (PFKFB3) is an enzyme that controls the intracellular concentration of fructose-2,6-bisphosphate which is an allosteric activator of 6-phosphofructo-1-kinase (PFK-1), a key enzyme of glycolysis. PFK-1 is tightly controlled by multiple metabolic feedback mechanisms and dictates the overall rate of glycolytic flux to lactate and the TCA cycle. In human cancers, several oncogenic proteins (e.g. HIF-1α, PTEN, and AKT) converge to increase the expression and activity of PFKFB3, leading to the high glycolytic rates typically observed in cancer cells. In addition to being a promising cancer metabolism target, PFKFB3 is required for the differentiation and tumor-promoting functions of the immunomodulatory Th17 cells and myeloid derived suppressor cells (MDSCs), which are attractive cellular targets to induce tumor immunity and potentially mediate intrinsic resistance to immune checkpoint inhibitors. PFK-158 is a potent selective small molecule inhibitor of PFKFB3 that displays broad anti-tumor activity causing significant growth inhibition in human and syngeneic preclinical models. As resistance mechanisms frequently activate pathways that result in up-regulation of glycolysis and PFKFB3, combination treatments of PFK-158 with cytotoxic and targeted agents have resulted in increased efficacy and tumor regression. Results: PFK-158 is a potent selective small molecule inhibitor of PFKFB3 that displays broad anti-tumor activity and causes significant growth inhibition in multiple human and syngeneic preclinical models. The tolerability and potential clinical benefit of PFK-158 are being investigated in advanced cancer patients with solid malignancies in a Phase 1 dose-escalation, multi-center clinical trial (clinicaltrials.gov # NCT02044861). The final cohort (650 mg/m2) has been open for enrollment and PFK-158 has been well tolerated to date. Secondary end-points have been incorporated to assess peripheral F2,6BP levels and immunosuppressive and effector cells populations. Of the 15 patients evaluable for response assessment at the end of two months of treatment, 6 patients have experienced a clinical benefit associated with PFK-158 administration, including a late stage pancreatic cancer patient that had a 75% reduction in her CA19-9 levels after 1 month, a renal cell carcinoma patient currently in month 9 and an adenocystic carcinoma patient in month 12. In addition, we examined the immunomodulatory effects of PFK-158 on Th17 cells and MDSCs in vitro, in B16 melanoma-bearing mice and in advanced cancer patients and found that PFK-158: (i) suppresses human Th17 cell and MDSC differentiation in vitro; (ii) decreases splenic and tumor-infiltrating Th17 cells, γδ T17 cells and MDSCs, and increases CD4+ and CD8+ T cells in the tumors of B16-F10 melanoma-bearing mice; and (iii) decreases peripheral blood Th17 cells, γδ T17 cells and MDSCs and increases activated effector CD4+ and CD8+ T cells in advanced cancer patients. Interestingly, we are discerning a correlation between the initial level of circulating Th17 cells and clinical responses to PFK-158. Conclusion: PFK-158 is the first-in-human and first-in-class PFKFB3 inhibitor that is currently under clinical development. To date, PFK-158 has been well tolerated and shows signs of clinical activity. In addition to controlling glycolysis, over expression of PFKFB3 in key immunesuppressive cells also leads to an immunomodulatory mechanism of action, suggesting that additional clinical benefit could result from combining PFK-158 with targeted agents as well as with immunotherapeutic agents. Note: This abstract was not presented at the conference. Citation Format: Sucheta Telang, Kavitha Yaddanapudi, Jaspreet Grewal, Rebecca Redman, Siqing Fu, Paula Pohlmann, Devalingam Mahalingam, Michael Kurman, Gilles Tapolsky, Jason Chesney.{Authors}. PFK-158 is a first-in-human inhibitor of PFKFB3 that selectively suppresses glucose metabolism of cancer cells and inhibits the immunosuppressive Th17 cells and MDSCs in advanced cancer patients. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B90.

HIV persists in CCR6+CD4+ T cells from colon and blood during antiretroviral therapy.

Objectives:The objective of this article is to investigate the contribution of colon and blood CD4+ T-cell subsets expressing the chemokine receptor CCR6 to HIV persistence during antiretroviral therapy.Design:Matched sigmoid biopsies and blood samples (n = 13) as well as leukapheresis (n = 20) were collected from chronically HIV-infected individuals receiving antiretroviral therapy. Subsets of CD4+ T cells with distinct differentiation/polarization profiles were identified using surface markers as follows: memory (TM, CD45RA−), central memory (TCM; CD45RA−CCR7+), effector (TEM/TM; CD45RA−CCR7−), Th17 (CCR6+CCR4+), Th1Th17 (CCR6+CXCR3+), Th1 (CCR6−CXCR3+), and Th2 (CCR6−CCR4+).Methods:We used polychromatic flow cytometry for cell sorting, nested real-time PCR for HIV DNA quantification, ELISA and flow cytometry for HIV p24 quantification. HIV reactivation was induced by TCR triggering in the presence/absence of all-trans retinoic acid.Results:Compared with blood, the frequency of CCR6+ TM was higher in the colon. In both colon and blood compartments, CCR6+ TM were significantly enriched in HIV DNA when compared with their CCR6− counterparts (n = 13). In blood, integrated HIV DNA levels were significantly enriched in CCR6+ versus CCR6− TCM of four of five individuals and CCR6+ versus CCR6− TEM of three of five individuals. Among blood TCM, Th17 and Th1Th17 contributed the most to the pool of cells harboring integrated HIV DNA despite their reduced frequency compared with Th2, which were infected the least. HIV reactivation was induced by TCR triggering and/or retinoic acid exposure at higher levels in CCR6+ versus CCR6− TM, TCM, and TEM.Conclusion:CCR6 is a marker for colon and blood CD4+ T cells enriched for replication-competent HIV DNA. Novel eradication strategies should target HIV persistence in CCR6+CD4+ T cells from various anatomic sites.

Clinical Significance of the Balanceshift of Th1 and Th2 Type Cytokines in Patients with Primary Immune Thrombocytopenic Purpura Detected by Cytometric Bead Array

To investigate the levels of Th1/Th2 cytokines in peripheral blood of patients with primary immune thrombocytopenic purpura(ITP) before and after treatment and their clinical significance. Ninety-eight cases of ITP were treated with glucocorticoid(GC), then were divided into 2 groups: effectively treated group and uneffectively treated group according to efficacy of treatment, 40 healthy persons confirmed by health examination were selected and enrolled in control group. The levels of Th1 cytokines(IFN-γ,TNFα,IL-2) and Th2 cytokines(IL-4,IL-5,IL-10) were detected by cytometric bead array before and at 4 weeks, 3 and 6 months after treatment and the relationship among detected indexes was analyzed. Before treatment with glucocorticoid, the levels of Th1 type cytokines were in 98 patients with ITP were higher and the levels of Th2 type cytokines were lower, compared with the healthy controls(P<0.05). The IL-2/IL-4 ratio was significantly higher than that of healthy controls(P<0.05). After treatment, the levels of Th1 type cytokines in effectively treated group were significantly decreased and the levels of Th2 type cytokines were significantly increased, compared with level before treatment(P<0.05). The IL-2/IL-4 ratio was significantly decreased after treatment for 6 months, compared with that before treatment(1.05±0.43 vs 2.53±0.72)(P<0.05), but the level of Th1 or Th2 type cytokines did not obviously changed. Peripheral blood Th1 and Th2 cells express abnormally in ITP patients, ITP is a Th1 dominated disease; the change of IL-2/IL-4 ratio before and after treatment correlated with the prognosis of ITP patients, displaying clinical significance for ITP individual therapy.

High cord blood CCL22/CXCL10 chemokine ratios precede allergic sensitization in early childhood.

Atopic diseases are known to be characterized by a T helper (Th) 2-skewed immunity; however, there are few studies addressing the Th1/Th2 immunity at birth related to the development of atopic diseases in early childhood. We investigated 186 children followed up regularly at the clinic for 4 years in a birth cohort study. The Th1-associated CXC chemokine ligand (CXCL)-10, CXCL11, and the Th2-associated CC chemokine ligand (CCL)-17 and CCL22 were quantified in cord blood by multiplex Luminex kits. Specific immunoglobulin E antibodies against food and inhalant allergens were measured at 6 months as well as 1, 1.5, 2, 3, and 4 years of age. Cord blood CCL22 levels were positively associated with IgE sensitization at age 2, whereas cord blood CXCL10 levels were negatively associated with mite sensitization at age 3. Furthermore, a high cord blood CCL22/CXCL10 chemokine ratio was significantly associated with a higher risk of allergic sensitization at age 3 (OR, 1.02; 95% confidence interval [CI], 1.0051.039; P = 0.012). However, cord blood Th1- and Th2-associated chemokines and their ratios were not associated with atopic diseases at different age. Our study indicates that a Th2-skewed immunity at birth may increase risk of allergic sensitization but not of allergic outcomes later in life.

MicroRNA-155-induced T lymphocyte subgroup drifting in IgA nephropathy.

MicroRNA-155 (miR-155) is an important immune regulator of T lymphocyte subgroup balance and function. This study was performed to explore the relationships between miR-155 expression in peripheral blood mononuclear cells (PBMCs), T lymphocyte subgroups, T lymphocyte regulators, and the clinical manifestations of IgA nephropathy (IgAN) patients. Sixty biopsy-proven IgAN patients and 25 healthy controls were included in this study. The expression of miRNAs in PBMCs was determined using a microRNA microarray and real-time RT-PCR. T lymphocyte subgroups (Th1, Th2, Treg, and Th17), differentiation regulators (c-Maf, STATA-6, GATA-3, SOCS-1, and Fxop3), cytokines (IFN-γ, IL-5, IL-10, and IL-17), serum IgA1 glycosylation level, and Cosmc expression were examined using flow cytometry, qPCR, and ELISA. Microarray analysis and qPCR suggested that the miR-155 level in PBMCs from IgAN patients was significantly lower than that in healthy controls (decreased 61%, 0.197±0.068 vs 0.796±0.13, p<0.01). The expression of GATA-3 (0.08±0.02 vs 0.04±0.01, p=0.035), SATA-6 (0.12±0.02 vs 0.06±0.01, p=0.036), and SOCS-1 (0.04±0.01 vs 0.03±0.01, p=0.01) was significantly higher in IgAN PBMCs compared to healthy controls, while that of Foxp3 (0.013±0.003 vs 0.040±0.01, p=0.026) and Cosmc (0.08±0.02 vs 0.19±0.04, p=0.024) was remarkably lower. Flow cytometric analysis revealed that the percentages of peripheral blood Th1 (17.35±1.22 vs 20.89±1.22, p=0.042) and Treg cells (1.86±0.15 vs 2.48 vs 0.26, p=0.031) were significantly lower in IgAN patients than in normal controls; however, the percentages of Th2 (1.73±0.29 vs 1.04±0.18, p=0.046) and Th17 (4.09±0.45 vs 2.06±0.21, p<0.001) were remarkably higher. ELISA results indicated that serum Th1 cytokine INF-γ (40.77±8.39 vs 83.02±17.4pg/mL, p=0.035) and Treg cytokine IL-10 (0.77±0.28 vs 4.18±1.34pg/mL, p=0.02) levels were lower, while Th2 cytokine IL-5 (1.02±0.17 vs 0.65±0.07pg/mL, p=0.04) and Th17 cytokine IL-17 (53.78±12.20 vs 28.87±4.59pg/mL, p=0.05) were significantly higher in IgAN patients than in normal controls. Significant correlations were found between miR-155 levels and Foxp3, Cosmc level, 24-h urine protein amount, urine RBC count, serum IgA concentration, and IgA1 dys-glycosylation level. A remarkably lower expression of peripheral lymphocyte miR-155 was observed in IgAN patients, leading to T lymphocyte subgroup drifting (increases in Th2 and Th17 along with decreases in Th1 and Treg), which inhibits Cosmc gene expression and worsens the aberrant glycosylation of IgA1 in IgAN patients. These results suggest that miR-155 may play an important role in the pathogenesis of IgAN and could serve as a potential disease biomarker.

Peripheral blood T helper type 17 frequency shows an inverse correlation with disease activity and magnetic resonance imaging-based osteitis and erosions in disease-modifying anti-rheumatic drug- and steroid-naive established rheumatoid arthritis.

An increased expansion of T helper type 17 (Th17) cells in the synovium has been shown to play a key role in cartilage and bone destruction in rheumatoid arthritis (RA). Because the correlation of the peripheral blood helper T cell subsets and various inflammatory cytokines with the magnetic resonance imaging (MRI)-based parameters have not been studied adequately to date, we sought to look for the same in this study. RA patients with disease duration less than 36 months, disease-modifying anti-rheumatic drugs (DMARDs) and steroid-naive, were recruited. MRI of the dominant hand and wrist was performed using a 0·2 Tesla MRI machine. Peripheral blood Th1 and Th17 were enumerated by flow cytometry and serum interleukin (IL)-6 and IL-17 by enzyme-linked immunosorbent assay (ELISA). Forty consecutive seropositive RA patients [33 females, mean disease duration 12·2 months, mean disease activity score (DAS)28 = 4·4] were included. MRI revealed erosions in 80% of these subjects. On subgroup analysis, prevalence of erosions (94 versus 68%) as well as mean erosion score (11·5 ± 18·9 versus 3·5 ± 6·0) were significantly higher in established RA (13-36 months' duration) compared to early RA (0-12 months). The median peripheral blood Th17 frequencies were significantly higher in patients (1·4%) compared to healthy controls (0·7%) and had a strong negative correlation with MRI parameters of erosion and osteitis as well as with DAS28 in the established RA subgroup. The frequency of peripheral blood Th17 subset was significantly expanded in established RA which correlated inversely with disease activity as well as MRI based erosions and osteitis.

Study on the effect and mechanism of the dysfunction of CD4(+) T cells in the disease process of chronic cardiac failure.

To study the effect and mechanism of the dysfunction of CD4(+) T cells in the disease process of chronic cardiac failure (CHF). According to different group technologies, 100 CHF patients were divided into the following groups: ischemia group and non-ischemia group, heart function Ⅲ-Ⅳ group and heart function Ⅰ-Ⅱ group, event group and non-event group, and 50 healthy volunteers were included in the control group. Real-time PCR was used to detect transcription factors T-bet and GATA-3 of Th1 and Th2; flow cytometry was applied to determine the ratio of Th17 and Treg cells; ELISA was employed to test cytokines IFN-γ, IL-4, IL-17 and IL-10 of peripheral blood Th1, Th2, Th17 and Treg cells, respectively; ultrasonic cardiogram was used to exploit to LVEF and LVEDd; and electrochemilu minescene immunoassay was used to examine plasma BNP. The differences of all indexes of all groups were analyzed and the correlation between CD4 T cells and clinical indexes was analyzed by Pearson correlation analysis. As compared to the control group, the transcription factors T-bet and GATA-3 of Th1 and Th2, the ratio of cytokines Th17 and IFN-γ, cytokines IL-17, T-bet/GATA-3, IFN-γ/IL-4, Th17 cells/Treg cells, IL-17/IL-10 of the ischemia group and non-ischemia group, heart function Ⅲ-Ⅳ group and heart function Ⅰ-Ⅱ group, event group and non-event group were all increased significantly, while their transcription factor GATA-3 of Th2, cytokines IL-4, Treg cells ratio, cytokines IL-10 were decreased obviously. The differences showed statistical significance (P<0.05). The increase or decrease of the partial CD4+ T cells of the ischemia group, heart function Ⅲ-Ⅳ group and event group was more distinctly. The results of Pearson correlation analysis showed that IFN-γ and IL-17 were significantly positively correlated with LVEDd and BNP, IL-4 and IL-10 were also significantly positively correlated with LVEF, but correlated negatively with BNP, and IL-17 was negatively correlative with LVEF. There was a correlation between CHF and the dysfunction of CD4(+) T cells showing immune activation phenomenons of deviations from the Th1/Th2 balance towards Th1 and from the Th17/Treg balance towards Th17, which was also related to the types, severity and prognosis of the disease.

IL-17A-producing T cells are associated with the progression of lung adenocarcinoma.

Accumulating evidence has shown that T cells are crucial in shaping the tumor microenvironment and regulating tumor development. However, the roles of IL-17A-producing T cells (IL-17A+CD4+ Th17, IL-17A+CD8+ Tc17 and IL-17A+ γδT17 cells) and related cytokines in the progression of lung cancer (LC) remain uncertain. Here, we found that the frequencies of both Th17 and γδT17 cells in the peripheral blood of patients with lung adenocarcinoma (LA) were higher than those in healthy controls (HCs), whereas the frequency of Tc17 cells in the patients with LA was decreased. In addition, the frequencies of circulating Th17 and γδT17 cells, but not Tc17 cells, were positively associated with tumor invasion and metastasis. Furthermore, the major source of IL-17A production was Th17 cells, followed by Tc17 and γδT17 cells, in peripheral blood from patients with LA and HCs; but the percentages of Th17 and γδT17 cells in total intracellular IL-17A+ cells obtained from the patients with LC were higher than those from HCs. Moreover, the protein and corresponding mRNA levels of IL-17A, IL-23, IL-1β, and TGF-β1 were much higher in the patients with LA than those in HCs, and the levels of IL-17A in patients were positively correlated with numbers of both Th17 and γδT17 cells, but not Tc17 cells. Finally, the frequencies of circulating Th17 and γδT17 cells, along with the levels of IL-17A, IL-23, IL-1β, and TGF-β1 were decreased in the patients with LA after tumor resection, whereas the frequency of circulating Tc17 cells was inversely increased in these patients. Our findings indicate that Th17, Tc17, γδT17 cells, and IL-17A-associated cytokines contribute to the development of LA and thus represent promising targets for therapeutic strategies.

Dietary ω-6/ω-3 Polyunsaturated Fatty Acid Ratios Affect the Homeostasis of Th/Treg Cells in Mice With Dextran Sulfate Sodium-Induced Colitis.

This study evaluated the effect of different dietary ω-6/ω-3 polyunsaturated fatty acid (PUFA) ratios on modulating helper T (Th) and regulatory T (Treg) lymphocytes in mice with dextran sulfate sodium (DSS)-induced colitis. There were 3 control and 3 colitis groups. Mice were fed for 24 days with diets with soybean oil (S), a mixture of soybean oil and low fish oil content (LF), or high fish oil content (HF). The ratio of ω-6/ω-3 PUFA in the LF diet was 4:1, and that in the HF diet was 2:1. The control groups drank distilled water while colitis groups were provided 2% DSS in drinking water during days 15-19. All mice drank distilled water from days 20-24 for recovery and were sacrificed on day 25. Colitis resulted in higher blood Th1, Th2, and Th17 and lower Treg percentages. Also, plasma haptoglobin and proinflammatory chemokines were elevated in colon lavage fluid. Colitic groups with fish oil had lower inflammatory mediators in the plasma and colon lavage fluid. Furthermore, the percentages of blood Th1, Th2, and Th17 cells were lower, whereas Treg cell percentages were higher than those in the soybean oil group. The colitis group with an ω-6/ω-3 PUFA ratio of 2:1 had more pronounced effects than the group with a ratio of 4:1. Diets with an ω-6/ω-3 PUFA ratio of 2:1 or 4:1 regulate the Th/Treg balance and attenuate inflammatory mediator production in colitis. Compared with the ω-6/ω-3 PUFA ratio of 4:1, the ratio of 2:1 was more effective in reducing inflammatory reactions in DSS-induced colitis.

BCL-2 protects human and mouse Th17 cells from glucocorticoid-induced apoptosis

Glucocorticoid resistance has been associated with Th17-driven inflammation, the mechanisms of which are not clear. We determined whether human and mouse Th17 cells are resistant to glucocorticoid-induced apoptosis. Freshly isolated human blood Th17 cells and in vitro differentiated Th17 cells from IL-17F red fluorescent protein reporter mice were treated with dexamethasone, a potent glucocorticoid. Apoptosis was measured using annexin V and DAPI staining. Screening of apoptosis genes was performed using the apoptosis PCR array. Levels of molecules involved in apoptosis were measured using quantitative RT-PCR, flow cytometry, and Western blotting. Knockdown of BCL-2 in murine Th17 cells was performed via retroviral transduction. Cytokines were measured using ELISA. A murine Th17-driven severe asthma model was examined for Th17 glucocorticoid sensitivity in vivo. Human and mouse Th17 cells and mouse Th2 cells were resistant to glucocorticoid-induced apoptosis. Th17 cells had glucocorticoid receptors levels comparable to those in other T effectors cells. Th17 cells had high levels of BCL-2, knockdown of which sensitized Th17 cells to dexamethasone-induced apoptosis. Production of IL-22, but not IL-17A and IL-17F, was suppressed by glucocorticoids. STAT3 phosphorylation in Th17 cells was insensitive to glucocorticoid inhibition. Lung Th17 cells in the murine severe asthma model were enhanced, rather than suppressed, by glucocorticoids. Th17 cells are resistant to glucocorticoid-induced apoptosis and cytokine suppression, at least in part due to high levels of BCL-2. These findings support a role of Th17 cells in glucocorticoid-resistant inflammatory conditions such as certain endotypes of asthma.

Imbalance of peripheral blood Th17 and Treg responses in children with refractory Mycoplasma pneumoniae pneumonia.

Mycoplasma pneumoniae (MP) pneumonia is usually a benign self-limiting disease, but some patients suffer from acute lung injury in MP infections who need to use immune-modulators with conventional antibiotic treatment. The imbalance between CD4+CD25+Foxp3+regulatory T (Treg) cells and T helper (Th17) cells has been found in a number of different inflammation and autoimmune disease, while the role of the Th17/Treg balance in MP pneumonia remains largely unknown. The aim of this study was to investigate the Th17/Treg pattern and its impact on disease severity in patients with MP pneumonia. This study involved 36 healthy control and 83 patients including 25 patients with refractory MP pneumonia and 58 patients with macrolide responsive MP pneumonia. The percentages of circulating Th17 cells and Tregs were determined by flow cytometry. The concentrations of serum Th17- and Treg-related cytokines were measured by enzyme-linked immunosorbent assay (ELISA). In comparison with that in the patients with macrolide responsive MP pneumonia and healthy control, significantly higher frequencies of Th17 cells and higher levels of IL-17 were detected in patients with refractory MP pneumonia. But there are no significant difference with the frequencies of Tregs and the levels of TGF-β1 in the patients with refractory MP pneumonia and patients with macrolide responsive MP pneumonia. However the Th17/Treg ratio was found to be significantly higher in the patients with refractory MP pneumonia than the patients with macrolide responsive MP pneumonia. An imbalance of circulating Th17 cells and Tregs is associated with the deterioration of pulmonary injury in patients with MP infections.

Change of Th17 Lymphocytes and Treg/Th17 in Typical and Atypical Optic Neuritis.

BackgroundTypical and atypical optic neuritis (ON) are two clinical types of autoimmune inflammatory diseases of the optic nerve that causes acute vision loss, and are difficult to distinguish in their early stages. The disturbance in the balance of Th17 and Treg lymphocytes is thought to play an essential role in these autoimmune inflammatory diseases.ObjectivesTo detect the clinical relevance of Th17 and Treg in peripheral blood and the ratio of Treg/Th17 in patients with typical and atypical ON. To determine whether analysis of Th17 and Treg lymphocytes will provides insights into the different disease phenotypes of typical and atypical ON.MethodsWe studied a consecutive series of patients aged 14–70 years who presented to our neurological department with typical ON (n = 30) or atypical ON (n = 33) within 4 weeks of their acute attacks. Routine clinical tests and ophthalmological examination were performed in all patients. Blood samples were collected from untreated patients and from gender- and age-matched healthy controls (n = 30). The proportion of peripheral blood Th17 cells and Treg cells was determined by flow cytometry.ResultsPatients with atypical ON had a higher proportion of Th17 cells than patients with typical ON (3.61±1.56 vs 2.55±1.74, P<0.01) or controls (1.45±0.86, P<0.01). The proportion of Th17 cells in patients with typical ON was also markedly higher than in controls (P<0.01). The mean percentage of Treg cells in atypical ON (6.31±2.11) and typical ON (6.80±2.00) were significantly lower when compared to controls (8.29±2.32, both P<0.01). No significant difference in Treg frequency was observed between typical ON and atypical ON (p>0.05).ConclusionsThe frequency of Th17 cells is higher in atypical ON than typical ON, and patients with atypical ON have a greater imbalance of pro-inflammatory and regulatory cells than patients with typical ON when compared with controls. These changes are indicative of distinct pathological mechanisms and may provide useful information to distinguish typical and atypical ON.