Effect of mechanical stimulation by rubbing abdomen by interfering with Th17 cell function on intestinal homeostasis in IBS-C model mice / 国际生物医学工程杂志

Objective:To investigate the possible mechanism of rubbing abdomen to regulate intestinal homeostasis in irritable bowel syndrome with constipation (IBS-C) mouse models.Methods:IBS-C mouse models of intestinal immune dysfunction were established using trinitrobenzene sulfonic acid (TNBS)-induced C57BL/J6 male mice. Thirty C57BL/J6 mice were randomly divided into three groups: the control group, the model group, and the mogul group. After 7 days of modeling, mice in the mogul group were given a mogul mechanical stimulation intervention once per day for 2 weeks, while mice in the control and model groups were not given any intervention. Changes in serum levels of interleukin-6 (IL-6) and IL-17A were detected by enzyme-linked immune sorbent assay (ELISA) and immunohistochemistry. The gene expression and protein levels of IL-17A and IL-23 were detected by quantitative real-time fluorescence PCR (qRT-PCR) and Western Blot, respectively. The morphological changes were observed by HE staining. The CD44 and CD62L expression changes were observed by immunofluorescence staining.Results:Compared with the model group, the levels of IL-6 and IL-17A in the serum of mice in the mogul group were decreased, and the expression of IL-6 and IL-7A in the tissues was down-regulated (all P<0.001). In addition, the gene expression and protein expression levels of IL-17A and IL-13 in the tissues of mice in the mogul group were decreased (all P<0.001). HE staining results showed that the mogul mechanical stimulation intervention could repair colonic tissues and reduce the inflammatory response. Immunofluorescence staining results showed that mogul mechanical stimulation intervention could downregulate the expression of CD44 but had no modulating effect on the expression of CD62L. Conclusions:Rubbing abdomen can improve intestinal homeostasis in IBS-C model mice by regulating changes in Th17 cell function.

Effect of Asiaticoside on Expression of Th17/Treg Cells in DBA/1 Mice with Collagen-Induced Arthritis / 中国实验方剂学杂志

ObjectiveTo observe the effect of asiaticoside （AC） on the expression of T helper 17 （Th17） cells and regulatory T （Treg） cells in DBA/1 mice with collagen-induced arthritis （CIA）. MethodMale SPF DBA/1 mice were randomized into six groups according to body weight： control group， CIA group， methotrexate group （MTX group， ip， 0.5 mg·kg-1）， and AC low-， medium-， and high-dose groups （ig， 5， 15， 45 mg·kg-1， respectively）. Modeling was performed in rats other than the control group. To be specific， they were immunized with bovine type Ⅱ collagen and complete Freund's adjuvant on the first day and with bovine type Ⅱ collagen and incomplete Freund's adjuvant on the 21st day. Administration began on the day of the second immunization， once a day for 28 days. On the 49th day， related tissues were collected. Then， hematoxylin-eosin （HE） staining was performed to observe the pathological changes of the joints. Immunohistochemical method was used to detect the expression of interleukin-17 （IL-17） and forkhead box protein-3 （FoxP3）， the markers of Th17 and Treg cells， respectively， immunofluorescence double staining the expression of IL-17 and FoxP3 in CD4+T cells of mouse joint tissue， and flow cytometry the proportions of Th17 and Treg cells in mouse lymph nodes. ResultCompared with the control group， CIA group demonstrated joint disorder， damage of articular cartilage and bone， severe bone erosion （P<0.01）， increase in stained CD4 and IL-17 and the integral absorbance （IA） （P<0.01）， decrease in stained FoxP3 and the IA （P<0.01）， rise of Th17/Treg ratio （P<0.01）， elevation of Th17 expression in mouse lymph nodes （P<0.01）， and reduction in Treg expression （P<0.01）. Compared with CIA group， MTX group and three AC groups showed normal joints， alleviated bone erosion and damage， intact and smooth joint surface， and decrease in stained IL-17 and IA （P<0.05， P<0.01）， and MTX group and AC medium-dose and high-dose groups registered decrease in stained CD4 and IA （P<0.01） and reduction in Th17/Treg ratio （P<0.05， P<0.01）. Moreover， AC medium-dose and high-dose groups showed rise in stained FoxP3 and IA （P<0.05， P<0.01）. In the lymph nodes of mice， decrease in expression of Th17 cells （P<0.05， P<0.01） and the increase in expression of Treg cells （P<0.05， P<0.01） were observed in all the three AC group. ConclusionAC can regulate Th17/Treg balance by inhibiting the expression of Th17 cells and promoting the expression of Treg cells in CIA mice.

Aging weakens Th17 cell pathogenicity and ameliorates experimental autoimmune uveitis in mice

Aging-induced changes in the immune system are associated with a higher incidence of infection and vaccination failure. Lymph nodes, which filter the lymph to identify and fight infections, play a central role in this process. However, careful characterization of the impact of aging on lymph nodes and associated autoimmune diseases is lacking. We combined single-cell RNA sequencing (scRNA-seq) with flow cytometry to delineate the immune cell atlas of cervical draining lymph nodes (CDLNs) of both young and old mice with or without experimental autoimmune uveitis (EAU). We found extensive and complicated changes in the cellular constituents of CDLNs during aging. When confronted with autoimmune challenges, old mice developed milder EAU compared to young mice. Within this EAU process, we highlighted that the pathogenicity of T helper 17 cells (Th17) was dampened, as shown by reduced GM-CSF secretion in old mice. The mitigated secretion of GM-CSF contributed to alleviation of IL-23 secretion by antigen-presenting cells (APCs) and may, in turn, weaken APCs' effects on facilitating the pathogenicity of Th17 cells. Meanwhile, our study further unveiled that aging downregulated GM-CSF secretion through reducing both the transcript and protein levels of IL-23R in Th17 cells from CDLNs. Overall, aging altered immune cell responses, especially through toning down Th17 cells, counteracting EAU challenge in old mice.

Study on the mechanism of Wenyang huazhuo tongluo formula contained serum inhibiting the proliferation of Th17 cells in scleroderma patients / 中国药房

OBJECTIVE To study the effects of Wenyang huazhuo tongluo formula conta ined serum on the expression and methylation of retinoic acid related nuclear orphan receptor （RORγt），and to explore the mechanism of its regulation of Th 17 cell proliferation in the treatment of scleroderma. METHODS Wistar rats were given 15，30，60 g/（kg·d）Wenyang huazhuo tongluo formula intragastrically to prepare different doses of drug-contained serum ，and peripheral blood of scleroderma patients were collected to sort Th 17 cells. Using blank serum as blank control ，methyltransferase inhibitor decitabine （DCA）as positive control ， Th17 cells were treated with different concentrations of drug-contained serum ，and then the proliferation of Th 17 cells was detected by CCK- 8；mRNA expressions of RORγt and IL-17 were detected by qRT-PCR ，and the protein expression of RORγt was detected by Western blot assay ；the protein expression of IL- 17 in the cell supernatant was detected by ELISA ，and the methylation level of RORγt gene promoter was detected by methylation-specific PCR. The transcriptional activity of RORγt gene promoter was detected by dual-luciferase assay. RESULTS Compared with blank control ，Wenyang huazhuo tongluo formula contained serum and DCA could inhibit the proliferation of Th 17 cells（P＜0.05），reduced the mRNA and protein expressions of RORγt and IL-17，enhanced the methylation level of RORγt gene promoter and attenuated ， the transcription activity of RORγt gene promoter. Except for mRNA expression of Th 17 and the methylation level of RORγt gene promoter in drug-contained serum low-dose group ，there were statistical significance in above indexes of other groups（P＜0.05）. CONCLUSIONS Wenyang huazhuo tongluo formula can promote the methylation lev el of RORγt gene，and inhibit the expression of RORγt and the secretion of IL-17，so as to inhibit the proliferation of Th 17 cells.

MicroRNA-10b Plays a Role in Bone Formation by Suppressing Interleukin-22 in Ankylosing Spondylitis

OBJECTIVE: The microRNA (miR)-10b is the T helper (Th) 17 cell specific in patients with ankylosing spondylitis (AS). The interleukin (IL)-22, which is closely related to Th17 cells, has been implicated in the regulation of new bone formation in experimental models. Therefore, the aim of this study was to evaluate whether miR-10b affects bone formation via the IL-22 pathway in AS.METHODS: Primary CD4+ T cells from AS were purified and transfected with miR-10b, anti-miR-10b, or scramble. Cell-surface markers and cytokine expression were analyzed by flow cytometry and enzyme-linked immunosorbent assay. Primary bone-derived cells (BdCs) from the facet joints of the spine were isolated, then osteogenic differentiation of primary BdCs was performed. We assessed alkaline phosphatase (ALP) activity and staining of BdCs at early time points. Alizarin red S staining of BdCs was performed at late time points.RESULTS: Overexpression of miR-10b reduced both IL-22 producing cell frequencies and cytokine production in T cells from the patients with AS. The IL-22 significantly increased ALP staining and bone mineralization. The ALP promotor activity of AS-BdCs was notably higher for the IL-22 concentration. The supernatants of the miR-10b overexpression group suppressed ALP activity on osteogenic progenitor cells from the facet joints of the spine in patients with AS.CONCLUSION: Our data suggest that miR-10b suppresses IL-22 production, which was involved in osteogenic proliferation in AS. Therefore, miR-10b might be a potential therapeutic candidate for regulation of new bone formation in patients with AS.

Advances in mechanisms for psoriasis and drug regulation / 药学学报

Psoriasis, bringing great trouble to patients' life, is a common chronic immune skin disease which is relapsed and difficult to cure. Further understanding of the pathogenesis of psoriasis will be benefit for potential drug targets and the development of therapeutic drugs. In recent years, "interleukin-23/T helper17 (IL-23/Th17) cell axis" has attracted more and more attention in the pathogenesis of psoriasis. Some drugs targeting the cell axis have also been successfully listed in the market to improve psoriasis and achieved good results. This review focuses on the "IL-23/Th17 cell axis" system to elaborate the role of a variety of immune cells involved in the pathogenesis of psoriasis, such as dendritic cells, macrophages, neutrophils and T cells, and summarizes multiple therapeutic antibodies targeting this cell axis in the treatment of psoriasis. In addition, this review also summarizes the current small molecule drugs for the treatment of psoriasis.

Mannan-binding lectin regulates Treg/Th17 cell balance in mice with Candida albicans infection / 中华微生物学和免疫学杂志

Objective To investigate the regulation of Treg/Th17 axis by mannan-binding lectin (MBL) in mice with Candida albicans ( C. albicans ) infection. -ethods MBL gene-knockdown (MBL-/-) C57BL/6 mice and wild-type (WT) C57BL/6 mice were divided into four groups. C. albicans strains (2×107 CFU) were injected intraperitoneally into the mice of infection groups. Paraffin sections of mouse liver and kidney tissues were prepared on 3 d. Histopathological changes were observed with hematox-ylin and eosin ( HE) and Periodic acid-Schiff ( PAS) staining. Flow cytometry was performed to analyze Th17 and Treg cells in mice on 7 d. The levels of IL-10 and IL-17A were measured by ELISA. CD4+T cells were obtained from spleen cells by magnetic sorting. Expression of Foxp3 and RORγt at mRNA and protein levels were detected by qRT-PCR and Western blot, respectively. Results The mouse model of C. albicans infection was established successfully. After infection, the MBL-/- mice had higher percentages of Th17 cells, but lower percentages of Treg cells than the WT mice. ELISA results showed that compared with the WT mice with C. albicans infection, the MBL-/- mice had significantly increased IL-17A and decreased IL-10 after infection. Moreover, the expression of RORγt at both mRNA and protein levels was up-regula-ted, while that of Foxp3 was down-regulated in the MBL-/- mice than in the WT mice following infection. Conclusions MBL regulates the immune balance of Treg/Th17 cells in mice infected with C. albicans through promoting the differentiation of CD4+ T cells into Treg cell subsets and inhibiting the differentiation into Th17 cell subsets.

Expressions of Treg/Th17 cells in post liver transplantation acute rejection in pediatric recipients diagnosed with biliary atresia / 中华肝胆外科杂志

Objective To study the expressions of regulatory T cell (Treg) and T helper cell 17 (Thl7) in post liver transplantation (LT) acute rejection in pediatric recipients diagnosed with biliary atresia.Methods 12 pediatric recipients with post-LT acute rejection in Tianjin First Center Hospital from July 2016 to April 2017 were included in the rejection group,and 22 patients with normal graft functions for more than 1 year post-LT were included in the stable group.The pre-LT primary disease in all the recipients was biliary atresia with cholestatic cirrhosis.Ten healthy children were included in the control group.Peripheral blood samples were taken before and at 1 week after anti-rejection treatment in the rejection group.Blood samples were taken during follow-up in the stable group and in the normal control.The proportions of Treg cell and Th17 cell were detected by flow cytometry.Results The proportions of Treg cells in the rejection group and the stable group were both significantly lower than the control group (P＜0.05).There was no significant difference between the rejection group and the stable group (P＞0.05).The proportion of Th17 cells in the rejection group was significantly higher than the stable group (P＜0.05) and the control group (P＜ 0.05),respectively;and the proportion in the stable group was significantly higher than the control group (P＜0.05).Meanwhile,there was also significant increase in the proportion of CD3+CD8+CD28+T in the rejection group (P＜0.05).After anti-rejection therapy,the proportions of Treg cell and Th17 cell became significantly lower than before therapy (P＜0.05).Conclusions The balance in Treg/Th17 cells occurred in the acute phase of rejection post-LT in pediatric recipients with biliary atresia,which persisted into the early period after anti-rejection therapy.

Clinical Efficacy of Shensuyin in Treating Adult Viral Myocarditis with Syndrome of Insufficiency of Lung-Qi / 中国实验方剂学杂志

Objective: To investigate the efficacy of Shensuyin in treating viralmyocarditis (VMC) with syndrome of insufficiency of lung-qi and the effect on levels of Th17 and Treg cells and relevant factors. Method: One hundred-four VMC cases were regaded as object of study and randomly divided into control group and observation group, with 52 cases in each group. Control group was treated with routine therapy by reference to ‘Chinese Guidelines for Diagnosis and Treatment of Heart Failure 2014’. In addition to the therapy of control group, observation group on the basis of treatment in the control group with Shensuyin, 1 dose/d, bid. One course of treatment was 8 weeks for both groups. Scores of Shensuyin, serum levels of Troponin I (cTnI) and cardiac free fatty acid binding protein (H-FABP), heart function and total efficacy were compared for both groups. Flow cytometry was used to detect peripheral blood levels of Th17 and Treg cells for the two groups. Serum levels of interleukin (IL) -17, IL-21, IL-10 were detected in both groups. Result: After treatment, scores of syndrome of Yin and Yang deficiency (shortness of breath, fatigue, dull chest pain, bad breath, cough) of observation group were obviously lower than those of control group (PPPPPConclusion: In addition to the routine therapy of VMC, the efficacy of Shensuyin has a significant effect in treating VMC with syndrome of lung qi deficiency, and the regulatory effect on levels of Th17 and Treg cells and relevant factors may be one of the effective ways.

Clinical Efficacy of Taohua Tang and Buzhong Yiqi Tang in Treatment of Deficiency Syndrome of Crohn's Disease / 中国实验方剂学杂志

Objective:To observe clinical efficacy of Taohua Tang and Buzhong Yiqi Tang on Crohn's disease (CD) at active phase (deficiency-cold in spleen and stomach), in order to observed its effect on Th1 and Th17 cytokines. Method:According to random number table, 86 patients with CD were divided into control group (42 cases) and observation group (44 cases). The control group (mild) was given SASP, 3-4 g·d-1, Po, tid. The control group (moderate or poor efficacy of SASP) was given prednisone acetate, 0.75 mg·kg-1·d-1, Po, tid. Observation group was given Taohua Tang and Buzhong Yiqi Tang in addition to therapy of the control group, 1 dose·d-1. The course of treatment was 12 weeks. Before and after treatment, Best CDAI, SES-CD, IBDQ and deficiency syndrome were scored, and levels of CRP, ESR, ALB, HB, PLT, IFN-γ, TNF-α, IL-2 and IL-17 were measured before and after treatment. Result:After treatment, the effect of traditional Chinese medicine(TCM) syndromes in the observation group was better than that in the control group (Z=2.058, PPPPZ=2.112, PZ=2.288, PPPγ, TNF-α, IL-2 and IL-17 levels in the observation group were lower than those in the control group (PConclusion:In addition to the therapy of conventional western medicine, Taohua Tang and Buzhong Yiqi Tang in treatment of deficiency syndrome of Crohn's disease (CD) can control the activity degree of the disease, reduce the degree of illness and inflammation, and improve the remission rate and the quality of life, with a better clinical efficacy than the pure western medicine therapy.

Primary study on the effect of glucose on mouse CD4+ T cell differentiation and its mechanism / 上海交通大学学报（医学版）

Objective :To study the effect of glucose on mouse CD4+ T cell differentiation. Methods :Mouse naïve CD4+ T cells cultured in the regulatory T cell (Treg), Th1, Th17 or Th2 differention condition were treated with different concentrations of glucose for 5 days. Treg, Th1, Th17 or Th2 percentages were measured by flow cytometry. Quantitative real-time PCR was used to detect the gene expressions of related cytokines and transcriptional factors. Results :The proportions of Treg and Th2 as well as the gene expressions of transforming growth factor-β, interleukin-4 (IL-4) and IL-13, and transcriptional factors, Foxp3 (forkhead box P3) and Gata3 (GATA binding protein 3), were increased significantly with the treatment of increasing concentration of glucose. On the contrary, with the glucose treatment, the percentages of Th1 and Th17 were reduced, and the gene expressions of the related cytokines and cytokine receptors, such as interferon-γ, IL-17A, IL-17F, IL-22 and IL-23R, and the related transcriptional factors, Tbx21 (T-box transcription factor 21) and RORC (RAR related orphan receptor C), were decreased consistently. Conclusion :Glucose promotes Treg and Th2 differentiation while inhibits Th1 and Th17 differentiation in vitro.

Exclusive enteral nutrition's moderating effect on Th17 cell of juvenile rat with inflammatory bowel disease / 国际儿科学杂志

Objective Through the TNBS-induced inflammatory bowel disease model in young rats,to observe the level of Th17 cell related cytokine IL-17A on the intestinal tissue of IBD young rats model after feeding EEN,Peptisorb,and investigate the enteral nutrition's regulative effects on Thl7 cells.Methods Thirty six four-week healthy male Sprague-Dswley rats were selected and divided into 4 groups,blank control group and EEN control group with 8 rats in each group,IBD group and IBD-EEN group with 10 rats in each group:blank control group and EEN control group were given normal saline enema,then fed with normal diet and enteral nutrition separately;IBD group and IBD-EEN group were given 5％ TNBS enema,establish the IBD model,then fed with normal diet and enteral nutrition separately.The weights,defecate characters and occult blood situations of all rats were determined.Also,we carried out the activity index score of disease according to the Cooper score method.On the seventh day,all rats were sacrificed and the colon specimens were collected.We observed mucosal injury by HE staining and expression level of Thl7 cell related cytokine IL-17A that were detected by immunohistochemical staining.Results Compared with IBD group,IBD-EEN group was improved significantly in terms of defecate character and occult blood situation.Both IBD group and IBD-EEN group gained their body weights than before,(39.95 ± 2.929) g and (38.40 ± 4.545) g respectively after 7 days (P≥0.05).In IBD group,gross tissue and pathological section displayed significant intestinal inflammation change,showing typical inflammatory bowel disease change;under the microscope,intestinal mucosa showed a large amount of inflammatory cell infiltration,a part of section displayed inflammatory granuloma formation,normal glandular structure was damaged.After EEN treatment,DAI scores werr decreased significantly;intestinal inflammation changes were relieved;intestinal mucosa inflammatory cell infiltration were reduced.Immunohistochemical study showed that the expression of IL-17A cytokine in the intestinal mucosa of IBD group was higher than that of IBD-EEN group (P ＜ 0.05).Conclusion The exclusive enteral nutrition treatment possessed the effect of reducing IBD young rat's intestinal mucosal inflammation,the inner mechanism may be related with the changing expression of Th17-cell cytokines,IL-17A.

The correlation between STAT3 phosphorylation and the activity of Behcet's disease / 复旦学报（医学版）

Objective To explore the role of signal transducer and activator of transcription 3 (STAT3) phosphorylation in the pathogenesis of Behcet's disease (BD),and to investigate the association between STAT3 phosphorylation and disease activity in BD patients.Methods Peripheral blood mononuclear cell (PBMC) was isolated from 15 mL peripheral boood of 10 active BD patients (BD-A),10 BD patients in remission (BD-R) and 10 healthy controls (HC) respectively.The blockade of STAT3 phosphorylation was performed by Stattic.The PBMC was divided into Stattic subgroup (treated with 2.5 μmol/L stattic and 1 640 medium,5 mL) and blank control subgroup (treated with 5 mL 1 640 medium),respectively.The protein levels of phosphorylated STAT3 (pSTAT3) and STAT3 were examined by flow cytometry and Western blot.The protein and mRNA levels of TNF-α,IFN-γ and IL-17 were tested by RT-PCR and ELISA.Two-way ANOVA and Bonferroni post hoc test were used to analyze the results.Results Compared with HC,the BD patients showed higher protein levels of pSTAT3 and STAT3,and higher protein and mRNA levels of TNF-α,IFN-γ and IL-17;compared with blank control subgroup,the protein levels of pSTAT3 and STAT3,and protein and mRNA levels of TNF-α,IFN-γ and IL-17 decreased in Stattic subgroup.In the BD-A group,the protein level of pSTAT3,and protein and mRNA levels of TNF-α,IFN-γ and IL-17 were significantly higher than those in the BD-R group.Conclusions An increased activation of the STAT3 pathway may contribute to the pathogenesis of BD and relate to disease activity in BD patients by inducing TH1 and Th17 cells activation.

Expression difference of Treg, Th17 cells and related cytokines between pre and post laparoscopic treatment of ovarian endometrioma / 中华内分泌外科杂志

Objective To investigate the changes and clinical significance of Treg and Th17 cells in patients with ovarian endometrioma (OEM) pre-and post-laparoscopy.Methods 36 patients with OEM undergoing laparoscopic surgery and confirmed by the pathology were enrolled as the experiment group,and 25 patients with fallopian tube obstruction who received laparoscopic examination were enrolled as the control group.The peripheral blood samples were collected from the control group and the experiment group before operation as well as from the experiment group 3 months after operation.Intraoperative peritoneal fluid were also collected from both groups.The levels of IL-17,IL-22,IL-23,IL-10 and TGF-β in serum and peritoneal fluid were detected by ELISA.The proportion of Treg and Th17 cells in peripheral blood was detected by flow cytometry.Results Compared with the control group,the levels of IL-17,IL-22 and IL-23 in the peritoneal fluid and serum of the experiment group increased significantly,while the levels of IL-10 and TGF-β were significantly decreased (P＜0.05).The proportion of Th17 cells in the peripheral blood of the experiment group increased significantly,and the proportion of Treg cells decreased significantly (P＜0.05).Compared with pre-laparoscopic treatment,the serum IL-17,IL-22 and IL-23,and the proportion of Th17 cells in peripheral blood were significantly decreased in the experiment group,and the levels of IL-10 and TGF-β in serum increased significantly (P＜0.05).Conclusions In patients with OEM,Treg and Th17 cell imbalance and the expression of related cytokines in disorder may be an important factor in the occurrence and development of disease.Detection of Treg/Th17 cells and their cytokines plays an important role in evaluating the severity of the disease and judging the efficacy of laparoscopic treatment and prognosis of OEM.

Role of Notch-Dll4 signaling pathway in autoimmune damage of Hashimoto thyroiditis / 中华内分泌代谢杂志

Forty patients with Hashimoto thyroiditis ( HT) and 20 healthy subjects with matched age-and sex-features ( NC) were selected. The patients with HT were further divided into normal thyroid function ( HT-A) and hypothyroidism ( HT-B) groups. Real-time PCR was performed to evaluate the expressions of Notch1, Dll4, and retinoid-related orphan receptor ( ROR )-γt mRNA. Flow-cytometry was used to detect the percentage of Th17 cells. Thyroid function, thyroid peroxidase antibody ( TPOAb) , and thyroglobulin antibody ( TgAb) were detected by electrochemiluminescence immunoassaies. The results showed that the Notch1, Dll4, ROR-γt mRNA levels and Th17 cell percentage were significantly increased in HT group compared with NC group (all P<0.01), especially in HT-B group. In HT patients, Notch1 and Dll4 mRNA expression levels were positively correlated with Th17 cell percentage and its transcription factor ROR-γt ( all P<0.01) . Besides, there were significantly positive correlations of Notch1 and Dll4 mRNA expressions with TPOAb and TgAb titers (P<0.05 or P<0.01). These results suggest that Notch1-Dll4 signaling pathway might be involved in the pathogenesis of thyroid-specific autoimmune damage by regulating Th17 cells in HT patients.

The role of intrahepatic regulatory T cells/helper thymphorytes 17 balance in patients with different phases of chronic hepatitis B virus infection / 中华传染病杂志

Objective To analyze the change of intrahepatic regulatory T cells (Treg )/helper thymphorytes (Th)17 balance in patients with different phases of chronic hepatitis B virus (HBV ) infection ,and to explore the role of Treg/Th17 balance in maintaining immune tolerance and inducing immune clearance ,and its influence on disease progression .Methods Sixty-eight patients with chronic HBV infection who underwent liver biopsy in Tianjin Second People′s Hospital were included .The 68 patients included 20 cases in immune tolerant (IT) phase ,36 cases in immune clearance (IC) phase and 12 cases in inactive phase .Eight healthy liver transplant donors were collected as healthy controls .The intrahepatic Treg/Th17 levels were detected by immuno-histochemical method . The changes of Treg/Th17 balance in patients with different phases of chronic HBV infection ,and the relationship between Treg/Th17 balance and the decreases of hepatitis B surface antigen (HBsAg ) , hepatitis B antigen (HBeAg) and HBV DNA levels in the peripheral blood were analyzed in patients with IC phase at two weeks of admission .Results The intrahepatic Treg and Th17 levels in IC phase group were the highest , then and they were higher in inactive phase group were higher than those in IT phase group ,And they were the lowest in control group .The Treg level in IC phase group increased significantly compared with the other three groups (all P< 0 .01) ,and there were no significant differences among the other three groups (all P> 0 .05) .The Th17 level between IT phase group and inactive phase group was not significantly different (P> 0 .05) ,while the differences were not significant in other groups (all P>0 .05) .Treg/Th17 ratio of IT phase group was the highest ,then the ratio of control group was higher than that of inactive phase group ,and IC phase group was the lowest ratio .The differences between IC phase group ,control group and IT phase group were significant (all P< 0 .05) ,and the difference between inactive phase group and IT phase group was also significant (all P<0 .05);and there was no significant difference among other groups (all P>0 .05) .The decreases of HBsAg ,HBeAg and HBV DNA levels in the peripheral blood at two weeks admission were negatively correlated with the intrahepatic Treg cell level in patients in IC phase of chronic HBV infection ( r= -0 .941 ,-0 .869 ,and -0 .883 ,respectively ,both P<0 .01) .The Treg ,Th17 levels and their ratio in IC phase group with different degree of inflammation and fibrosis had significant differences :G4 group > G3 group > G2 group ,S3 group > S2 group > S1 group (all P<0 .05) .Conclusions There is no change of the Treg/Th17 balance in IT phase ,and Treg has no influence on maintaining immune tolerance in chronic HBV infection .T he imbalance of Treg/Th17 is observed in IC phase .Th17 may actively participate in the immune-mediated liver injury and the development of hepatic fibrosis in CHB patients .Treg may inhibit inflammation and reduce liver injury via the negative feedback regulation mechanism ,and may impede the eradication of HBV simultaneously .

Role and clinical significance of imbalanced Th17/CD4+CD25+ regulatory T lymphocytes in children with Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis / 中华微生物学和免疫学杂志

Objective To investigate the changes in Th17 cells and CD4+CD25+regulatory T lym-phocytes ( Treg) as well as transcription factors and cytokines relating to them in children with Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) and to analyze their role and clinical significance. Methods Thirty-two children with newly diagnosed EBV-associated HLH in the Hematology/Oncology Department of Zhengzhou Children′s Hospital from January 2012 to December 2016 were enrolled in this study. Thirty healthy children taking physical examination in the same hospital in the corresponding period were recruited as controls. Percentages of Th17 and Treg cells in peripheral blood T lymphocytes were detected by flow cytometry. Expression of RORγt and Foxp3 at mRNA level in peripheral blood mononuclear cells was detected by real-time PCR. Levels of IL-6, IL-17, IL-10 and TGF-β1 in serum samples were measured by ELISA. Results Compared with the control group, the EBV-associated HLH group showed in-creased percentage of Th17 cells [(1. 09±0. 43)％ vs (0. 39±0. 19)％, P<0. 05] and enhanced expres-sion of RORγt at mRNA level [(1. 41±0. 37) vs (0. 67±0. 13), P<0. 05], but decreased percentage of Treg cells [(3. 66±1. 13)％ vs (6. 80±1. 15)％, P<0. 05] and inhibited expression of Foxp3 at mRNA level [(15. 97±5. 11) vs (30. 23±4. 95), P<0. 05]. All of the above mentioned changes were reversed af-ter treatment (P<0. 05). Serum levels of IL-6 and IL-17 of EBV-associated HLH group were higher than those of control group, while serum levels of IL-10 and TGF-β1 were lower (P<0. 05). Conclusion Im-balanced Th17/Treg cells might play an important role in the pathogenesis of EBV-associated HLH. Cyto-kines relating to the maintenance of Th17/Treg cell balance could be used as indicators of disease develop-ment.

Effects of mannan-binding lectin on differentiation of Th17 cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of mannan-binding lectin ( MBL) on the differen-tiation of Th17 cells (T helper cell 17, Th17). Methods CD4+T cells were separated in vitro from fresh human cord blood by MACS ( magnetic-activated cell sorting ) separator. Anti-CD3 monoclonal antibody ( McAb) and anti-CD28 McAb were used to stimulate CD4+T cells with IL-6 and TGF-β1 as inducers. Then, these cells were treated with ( MBL group) or without ( control group) different concentrations of MBL. Percentages of Th17 cells in different groups were detected by fluorescence-activated cell sorting( FACS) after 72 hours of culturing. Quantitative real-time PCR ( Q-PCR) was used to analyze the expres-sion of RORγt ( retinoid-related orphan receptor-γ-t) at mRNA level in both control and MBL groups. En-zyme-linked immunosorbent assay (ELISA) was performed to detect the levels of IL-17A in supernatants of cell culture from different groups. FACS was used to detect the percentages of Th17 cells in MBL-/-and wild type ( WT) mice. Results MBL could significantly reduce the percentage of Th17 cells after 72 hours of culturing as compared with the control group. Moreover, MBL could significantly down-regulate the expres-sion of RORγt at mRNA level and decrease the expression of IL-17A. Results of animal experiments showed that the percentages of CD4+RORγt+Th17 cells in MBL-/- mice were significantly higher than those in WT mice. Conclusion MBL can inhibit the differentiation of CD4+T cells to Th17 cells, which is induced by IL-6 and TGF-β1.

Role and clinical significance of imbalanced Th17/CD4+CD25+ regulatory T lymphocytes in children with Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis / 中华微生物学和免疫学杂志

Objective To investigate the changes in Th17 cells and CD4+CD25+regulatory T lym-phocytes ( Treg) as well as transcription factors and cytokines relating to them in children with Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) and to analyze their role and clinical significance. Methods Thirty-two children with newly diagnosed EBV-associated HLH in the Hematology/Oncology Department of Zhengzhou Children′s Hospital from January 2012 to December 2016 were enrolled in this study. Thirty healthy children taking physical examination in the same hospital in the corresponding period were recruited as controls. Percentages of Th17 and Treg cells in peripheral blood T lymphocytes were detected by flow cytometry. Expression of RORγt and Foxp3 at mRNA level in peripheral blood mononuclear cells was detected by real-time PCR. Levels of IL-6, IL-17, IL-10 and TGF-β1 in serum samples were measured by ELISA. Results Compared with the control group, the EBV-associated HLH group showed in-creased percentage of Th17 cells [(1. 09±0. 43)％ vs (0. 39±0. 19)％, P<0. 05] and enhanced expres-sion of RORγt at mRNA level [(1. 41±0. 37) vs (0. 67±0. 13), P<0. 05], but decreased percentage of Treg cells [(3. 66±1. 13)％ vs (6. 80±1. 15)％, P<0. 05] and inhibited expression of Foxp3 at mRNA level [(15. 97±5. 11) vs (30. 23±4. 95), P<0. 05]. All of the above mentioned changes were reversed af-ter treatment (P<0. 05). Serum levels of IL-6 and IL-17 of EBV-associated HLH group were higher than those of control group, while serum levels of IL-10 and TGF-β1 were lower (P<0. 05). Conclusion Im-balanced Th17/Treg cells might play an important role in the pathogenesis of EBV-associated HLH. Cyto-kines relating to the maintenance of Th17/Treg cell balance could be used as indicators of disease develop-ment.

Effects of mannan-binding lectin on differentiation of Th17 cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of mannan-binding lectin ( MBL) on the differen-tiation of Th17 cells (T helper cell 17, Th17). Methods CD4+T cells were separated in vitro from fresh human cord blood by MACS ( magnetic-activated cell sorting ) separator. Anti-CD3 monoclonal antibody ( McAb) and anti-CD28 McAb were used to stimulate CD4+T cells with IL-6 and TGF-β1 as inducers. Then, these cells were treated with ( MBL group) or without ( control group) different concentrations of MBL. Percentages of Th17 cells in different groups were detected by fluorescence-activated cell sorting( FACS) after 72 hours of culturing. Quantitative real-time PCR ( Q-PCR) was used to analyze the expres-sion of RORγt ( retinoid-related orphan receptor-γ-t) at mRNA level in both control and MBL groups. En-zyme-linked immunosorbent assay (ELISA) was performed to detect the levels of IL-17A in supernatants of cell culture from different groups. FACS was used to detect the percentages of Th17 cells in MBL-/-and wild type ( WT) mice. Results MBL could significantly reduce the percentage of Th17 cells after 72 hours of culturing as compared with the control group. Moreover, MBL could significantly down-regulate the expres-sion of RORγt at mRNA level and decrease the expression of IL-17A. Results of animal experiments showed that the percentages of CD4+RORγt+Th17 cells in MBL-/- mice were significantly higher than those in WT mice. Conclusion MBL can inhibit the differentiation of CD4+T cells to Th17 cells, which is induced by IL-6 and TGF-β1.