Wogonin promotes Th17/Treg cell balance in rat models with autoimmune hepatitis / 基础医学与临床

Objective To investigate the impacts of wogonin(WG)on Th17/Treg cell balance in autoimmune hep-atitis(AIH)rats.Methods A total of 10 rats were randomly selected as the control group.The remaining rats were injected with concanavalin A(ConA,12.5 mg/kg)solution via tail vein to construct AIH model rat,which were ran-domly divided into AIH group,L-WG group(10 mg/kg),M-WG group(20 mg/kg),H-WG group(30 mg/kg),H-WG+VPA group(30 mg/kg WG+300 mg/kg Notch signal pathway activator VPA),10 rats in each group and administered once a day for 10 days.Serum inflammatory factors and liver function indexes were detected by ELISA;HE staining was used to observe the pathological morphology of liver tissue;the level of spleen Th17/Treg cells was detected by flow cytometry;Western blot was used to detect the expression of spleen retinoid acid related orphan receptor γ t(RORγt),fork head box protein P3(Foxp3)and liver Notch signal pathway proteins.Results The liver tissue structure of control group was normal and the staining was clear;In AIH group,the cells of liver tis-sue showed edema,the increase of cell volume led to the compression and narrowing of liver sinuses,and a large number of inflammatory cell infiltration and a small amount of necrosis occurred.The contents of alanine aminotrans-ferase(ALT),aspartate aminotransferase(AST),interleukin(IL)-17 and IL-23,level of Th17 cells,Th17/Treg,the expression of RORγt,Notch,hes family bHLH transcription factor 1 gene(HES1)and hes related family bHLH transcription factor with YRPW motif 1(HEY1)protein in AIH group were greatly higher than those in control group(P＜0.05),the contents of IL-10 and TNF-β,level of Treg cells,and level of Foxp3 protein were greatly lower(P＜0.05);Compared with AIH group,the liver injury in L-WG group,M-WG group and H-WG group was im-proved,the contents of ALT,AST,IL-17 and IL-23,level of Th17 cells,Th17/Treg,the expression of RORγt,Notch,HES1 and HEY1 protein were greatly lower(P＜0.05),the contents of IL-10 and TNF-β,level of Treg cells,and level of Foxp3 protein were greatly higher(P＜0.05);VPA reversed the improvement effect of H-WG on AIH rats.Conclusions WG could promote Th17/Treg cell balance in AIH rats by down-regulating Notch signal pathway.

To Explore the Effect And Mechanism of Traditional Chinese Medicine on Chronic Urticaria Based on Th17/Treg Balance Axis / 世界科学技术-中医药现代化

Immune imbalance is believed to play a dominant role in the pathogenesis of chronic urticaria.While Th1/Th2 imbalance used to be considered as the main contributing factor of the development of chronic urticaria.Recently,Th17/Treg imbalance is found to be an important immune mechanism leading to the development of chronic urticaria.To be more specific,according to traditional Chinese medicine(TCM)'s comprehensive understanding of the etiology of chronic urticaria.it is generally believed that the pathogenesis of chronic urticaria is due to a lack of innate endowment,a lack of solidity of the body surface,and repeated exposure to six pathogenic factors.Another possible reason lies with dietary disorders that generate heat and wind or chronic illness and weakness and loss of nourishment of qi and blood.Therefore,in terms of the treatment,from the perspective of sthenia syndrome,it is advisable to remove the wind and disperse the pathogenic factors and clear the dampness and heat.From the perspective of asthenia syndrome,it is advisable to nourish the qi and blood and support the righteousness.As for mixed excess and deficiency,both support the healthy atmosphere and dispel the pathogenic factors are important.Regarding the effects of TCM on the balance of Th17/Treg in chronic urticaria and immune diseases,it mainly involved herbal compounding,herbal active ingredients and single herbs.However,the research attention has been drawn to investigating the role of TCM in the treatment of chronic urticaria and various immune diseases based on the research outcomes in modern pharmacological research.This can not only provide scientific evidence for TCM treatment of chronic urticaria,but also bring benefits to more patients with immune diseases.Therefore,the author reviews the recent research progress of TCM on the effects of Th17/Treg immune imbalance in chronic urticaria and other immune diseases by explaining the effects of Th17 and Treg cells in chronic urticaria.

Discussion on the Treatment of Thyroid associated ophthalmopathy from"Liver Opens at the Eyes"Based on the Imbalance of"Th17/Treg" / 世界科学技术-中医药现代化

Thyroid associated ophthalmopathy(TAO)is a refractory disease,which is related to Th17/Treg cellular immune imbalance."liver opens at the eyes"is a systematic summary of the theories of liver disease affecting the eye and eye disease affecting the liver in traditional Chinese medicine,which can guide the clinical diagnosis and treatment of TAO in traditional Chinese medicine.Studies have shown that the treatment of TAO from the perspective of"liver opens at the eyes"can regulate the immune imbalance of Th17/Treg cells to a certain extent,so as to achieve the purpose of disease prevention and treatment.This paper discusses the role of Th17/Treg cellular immune imbalance in TAO,from the aspects of Th17/Treg imbalance promoting the occurrence and development of TAO,the role of"liver opens at the eyes"theory in the etiology and pathogenesis of TAO,and the consistency between"liver opens at the eyes"physiology and Th17/Treg immune balance.In order to reveal the scientific connotation of traditional Chinese medicine in preventing and treating TAO by regulating Th17/Treg imbalance.

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.

Effect and Mechanism of Cxcr4 Gene-Modified BMSC-Derived Exosomes on Aplastic Anemia / 中国实验血液学杂志

OBJECTIVE@#To explore the improvement effect of CXC chemokine receptor 4 (Cxcr4) gene-modified bone marrow mesenchymal stem cell (BMSC)-derived exosomes on aplastic anemia (AA), and make a preliminary exploration of the mechanism.@*METHODS@#Mouse BMSCs were isolated and cultured, then infected by recombinant lentivirus carrying Cxcr4 gene. The expression of green fluorescence was observed through fluorescence microscope, the expression of Cxcr4 mRNA was detected by real-time fluorescence quantitative PCR, and the BMSC-derived exosomes modified with Cxcr4 gene were extracted. Mouse models of AA were constructed, and control group, model group (AA), AA+BMSC group, AA+NC-BMSC group, AA+Cxcr4-BMSC group were set up. Except control group and model group, the other three groups of mice were injected 400 μl exosomes from different sources via the tail vein, after 2 weeks, the routine blood indices and the number of bone marrow nucleated cells were detected, the pathological changes of bone marrow were observed by HE staining, and the expression level of Treg cells was detected by flow cytometry.@*RESULTS@#Mouse BMSCs were successfully isolated, and BMSCs with high expression of Cxcr4 and their exosomes were obtained. Compared with the control group, the number of red blood cell (RBC), white blood cell (WBC), and platelet (PLT), the hemoglobin (Hb) content and proportion of Treg cells in the peripheral blood of mice in the model group significantly decreased (P<0.01), as well as the number of bone marrow nucleated cells (P<0.01). The proliferation level of nucleated cells was low, and the medullary cavity was filled with a large number of fat cells. Compared with the model group, the number of RBC, WBC, PLT, the Hb content and proportion of Treg cells in the peripheral blood of mice in the AA+BMSC group, AA+NC-BMSC group, and AA+Cxcr4-BMSC group significantly increased (P<0.01), as well as the number of bone marrow nucleated cells (P<0.01), and pathological changes of bone marrow were improved. In addition, the number of RBC, WBC, PLT, the Hb content and proportion of Treg cells in the peripheral blood of mice in the AA+Cxcr4-BMSC group were significantly higher than those in the AA+BMSC group (P<0.01), as well as the number of bone marrow nucleated cells (P<0.01).@*CONCLUSION@#Injection of Cxcr4 gene-modified BMSC-derived exosomes has a certain improvement effect on AA mice, and the mechanism may be related to an increase of the proportion of Treg cells.

Th9, Th17, Treg Cells and their related cytokines in primary diagnosis of multiple myeloma / 中国输血杂志

【Objective】 To detect the abnormal expression of Th9, Thl7, Treg cells, interleukin-9 (IL-9), interleukin-17 (IL-17) and transforming growth factor-β(TGF-β) in patients with multiple myeloma (MM), and to explore their roles in primary diagnosis of MM. 【Methods】 The level of Th9, Th17 and Treg cells in peripheral blood of 54 MM patients with(patient) and 45 healthy volunteers (control) were measured by flow cytometry, and the levels of IL-9, IL-17 and TGF-β were detected by ELISA. 【Results】 The percentages(%) of Th9, Thl7 in MM patients increased significantly in comparison to controls [1.37±0.39 vs 0.79±0.26; 2.02±0.41 vs 1.18±0.32] (P<0.05). The proportion(%) of CCD4+ CD25+ FoxP3+ Treg cells in patients was significantly lower than those in controls (4.92±0.83 vs 7.04±1.85, P<0.05). The expression levels (%) of IL-9 and IL-17 in the peripheral blood of patients were significantly higher than those in controls (25.74 1±7.33 vs 16.82±5.58; 11.01±3.71 vs 7.68 ± 2.57, P<0.05). The levels of TGF-β in patients and controls were (3.73±1.44)% vs (6.95±2.12)%, showing a significant decrease (P<0.05). 【Conclusion】 The abnormal percentage of Th9, Thl7, Treg cells and the abnormal expression levels of IL-9, IL-17, TGF-β in MM patients may play an important role in the initial diagnosis of MM.

Klotho protein inhibits Treg/TH17 cell-mediated immune escape of cervical cancer cells by regulating TGF-β1/Foxp3/RORγt signaling pathway / 中国肿瘤生物治疗杂志

@#Objective: To study the effect of anti-aging Klotho protein on immune escape mediated by regulatory T cells (Treg)/helper T cells 17 (TH17) in mice bearing cervical cancer and its mechanism. Methods: The model of cervical cancer-bearing mice were established, and the control group (normal mice), model group (cervical cancer-bearing mice model), and Klotho treatment group (cervical cancer-bearing mice treated with Klotho protein, 200 ng/d) were set up. The weight of cervical cancer tumors in mice of each group was weighed at 7 and 14 days after treatment respectively, PBMCs were separated at the same time. Flow cytometry was used to detect the changes of T lymphocyte function and the proportion of Treg and TH17 cells in mice. qPCR was used to detect the expressions of Foxp3 and RORγt, the key transcription factors of Treg/TH17 cells, in PBMCs of mice in each group. The changes of IL-17, IL-6, IL10, TGF-β and IL-23 in PBMCs were detected by ELISA. The protein expressions of Klotho, TGF-β, Foxp3 and RORγt in PBMCs of mice were detected by WB assay. Results: On the 14th day, the tumor inhibition rate of the cervical cancer-bearing mice in the Klotho group was significantly higher than that in the Model group [(52.16±8.25)% vs (23.33±6.29)% the model group to be supplemented, P< 0.05). Compared with the Control group, the ratios of Treg and TH17 cells in the lymphocytes of the tumor-bearing mice significantly increased (all P<0.05), the ratios of total T lymphocytes (CD3+), auxiliary/induced T lymphocytes (CD3+CD4+) and immune index (CD3+CD4+/CD3+CD8+ cells) decreased significantly (all P<0.05); in addition, the mRNAexpressions of Foxp3 and RORγt genes, cytokines of IL-17, IL-6, IL-10, TGF-β and IL-23, as well as protein expressions of TGF-β1, Foxp3 and RORγt increased significantly (all P <0.05), while the level of Klotho protein significantly decreased in Model group (P<0.05). Compared with the Model group, the above indicators showed opposite changes in Klotho group (P<0.05), but there was no significant difference with the Control group (all P> 0.05). Conclusion: Klotho protein may inhibit Treg/TH17 cell-mediated immune evasion in cervical cancer-bearing mice by inhibiting TGF-β1/Foxp3/RORγt signaling pathway and exert anti-tumor effect.

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.

Blocking the PD-1/PD-L1 axis enhanced cisplatin chemotherapy in osteosarcoma in vitro and in vivo

BACKGROUND@#The blocking of the programmed cell death protein (PD-1)/programmed death-ligand 1 (PD-L1) axis has been found to have an anticancer activity against various types of cancer by enhancing T cell immunity, while there are no studies linking the PD-1/PD-L1 axis to chemotherapy drugs in osteosarcoma (OS). The present study aimed to investigate the effects of blocking PD-1/PD-L1 axis on the cisplatin chemotherapy in OS in vitro and in vivo.@*METHODS@#Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to detect PD-L1 mRNA in OS tissues. Cell proliferation and apoptosis were measured by Cell Counting Kit-8 (CCK-8) and flow cytometry assays, respectively. In vivo, the syngeneic mice were treated with cisplatin and anti-PD-1 antibody alone or jointly.@*RESULTS@#In this study, it revealed that PD-L1 mRNA was highly expressed in OS tissues. Further inhibitory evaluation showed that the K7M2-LV cells (PD-L1 overexpression) co-cultured with PD-1 lymphocytes could promote K7M2 cell proliferation. Meanwhile, the combination of anti-PD-1 antibody and cisplatin significantly decreased the proliferation and increased the apoptosis of K7M2 cells in a co-culture system. In vivo, the combination of anti-PD-1 antibody and cisplatin significantly inhibited tumor growth, while the mechanisms did not involve regulatory T cells.@*CONCLUSION@#The present data suggested that the blocking of PD-1/PD-L1 axis had a positive prognostic value, which can enhance the chemotherapeutic effect of cisplatin in OS. These findings provide a rationale for utilizing PD1/PD-L1 blocking antibodies as a single agent to cure refractory OS in patients receiving cisplatin treatment.

Effects of leptin on Treg cells and the possible mechanism / 中华微生物学和免疫学杂志

Objective To investigate the effects of leptin on Treg cells and the possible mecha-nism. Methods Leptin-deficient ( ob/ob) mice and homologous wild-type mice were used in this study. The percentages of Treg cells in spleen tissues and peripheral blood samples were measured by flow cytometry ( FCM) . Differences in Treg cell functionality were compared between the two groups. Splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by magnetic beads, were respectively cultured with leptin and anti-leptin neutralization antibody to evaluate the effects of leptin on Treg cells. Quantitative real-time PCR was performed to analyze the expression of Treg cell-related cytokines at transcriptional level. The levels of IL-10 and TGF-β in the supernatants of CD4+T cell culture were measured with Luminex technolo-gy. Results Compared with the wild-type mice, the ob/ob mice showed higher percentages of Treg cells in both peripheral blood samples and spleen tissues [(11. 56 ± 0. 72)％ vs (5. 47 ± 0. 81)％, (10. 16 ± 0.93)％ vs (6.29±0. 69)％]. Treg cells isolated from the ob/ob mice had stronger immunosuppressive effects on the proliferation of effector T ( Teff) cells and the secretion of TNF-α and IFN-γ than those from the wild-type mice [TNF-α:(1. 6±0. 2)％ vs (2. 4±0. 5)％, IFN-γ:(4. 3±0. 3)％ vs (7. 2±1. 2)％]. The percentages of Treg cells were decreased from (12. 2±1. 8)％ to (7. 6±0. 9)％ upon the in vitro treat-ment of CD4+ T cells from the ob/ob mice with leptin and the immunosuppressive effects of Treg cells were also weakened. However, the percentages of Treg cells were increased from (7. 8±0. 85)％ to (13. 1± 1. 5)％ upon the in vitro treatment of CD4+T cells from the wild-type mice with anti-leptin antibody and the immunosuppressive effects of Treg cells were improved as well. Moreover, the expression of Foxp3, IL-10 and TGF-β at transcriptional level and the levels of IL-10 and TGF-β in the ob/ob group were higher than those in the wild-type group. Conclusions Leptin deficiency significantly promoted the generation of Treg cells in mice and resulted in an increased expression of Foxp3, IL-10 and TGF-βat mRNA level and elevat-ed levels of IL-10 and TGF-β. The treatment of CD4+T cells with leptin might inhibit the generation of Treg cells through down-regulating the transcription of Foxp3, IL-10 and TGF-β.

Advances in the study of the mechanism of vitamin D treatment of psoriasis from the perspective of T cell immunity / 医学研究生学报

The pathogenesis of psoriasis is complex,including genetic,immune and many other factors,and Th cell play an important role in this process. Vitamin D excerts excellent therapeutic effect on psoriasis,and the therapeutic effect may be related to T cells. In this passage,we look into the recent advances in the pathogenesis of psoriasis from the perspectives of Th1/Th2,Th9,Th17,Treg and the advances in treatment of psoriasis with vitamin D,for providing some ideas and entry points for future research.

1,25-Dihydroxyvitamin D3 protects obese rats from metabolic syndrome promoting regulatory T cell-mediated resolution of inflammation

Vitamin D has been found to produce therapeutic effects on obesity-associated insulin resistance and dyslipidemia through its potent anti-inflammatory activity, but the precise immunomodulatory mechanism remains poorly understood. In the present study we found that 1,25-dihydroxyvitamin D [1,25(OH)D], the biologically active form of vitamin D, significantly attenuated monosodium glutamate (MSG)-induced obesity and insulin resistance as indicated by body weight reduction, oral glucose tolerance improvement, and a glucose infusion rate increase as detected with hyperinsulinemic-euglycemic clamp. Moreover, 1,25(OH)D not only restored pancreatic islet functions but also improved lipid metabolism in insulin-targeted tissues. The protective effects of 1,25(OH)D on glycolipid metabolism were attributed to its ability to inhibit an obesity-activated inflammatory response in insulin secretory and targeted tissues, as indicated by reduced infiltration of macrophages in pancreas islets and adipose tissue while enhancing the expression of in liver tissue, which was accompanied by increased infiltration of Treg cells in immune organs such as spleen and lymph node as well as in insulin-targeted tissues such as liver, adipose, and muscle. Together, our findings suggest that 1,25(OH)D serves as a beneficial immunomodulator for the prevention and treatment of obesity or metabolic syndrome through its anti-inflammatory effects.

Influences of interaction between DENV-2-infected HUVECs and regulatory T cells on major inflam-matory cytokines / 中华微生物学和免疫学杂志

Objective To investigate the influences on major inflammatory cytokines after co-cul-turing regulatory T cells (Treg) with human umbilical vein endothelial cells ( HUVECs) that were infected with dengue virus type 2 (DENV-2). Methods Peripheral blood mononuclear cells (PBMC) were extrac-ted from concentrated human leukocytes by density gradient centrifugation. Treg cells were sorted by immu-nomagnetic beads. Expression of CD4,CD25 and CD127 molecules on the membrane of Treg cells was detec-ted by flow cytometry to identify the purity of Treg cells. HUVECs pretreated with or without sphingosine-1-phosphate S1P type 1 (S1P1)-specific receptor agonist CYM-5442 for 24 h were first infected with DENV-2 and then co-cultured with Treg cells. Expression of IL-6,IL-8,TNF-α,IL-10 and TGF-β at mRNA level was detected by real-time RT-PCR. Levels of IL-6,IL-8,IL-10 and TGF-β in the culture supernatants were detec-ted by a double-antibody sandwich ELISA. Results The purity of Treg cells was (84. 3±0. 5)%. Expression of NS1 at mRNA level in DENV-2-infected HUVECs first gradually increased and then decreased after reac-hing the peak at 24 h (3. 03±0. 26, P<0. 01). Enhanced expression of IL-6,IL-8 and TNF-α at mRNA level in HUVECs was observed after DENV-2 infection ( P<0. 01). Expression of these cytokines at every time point was decreased after co-culturing DENV-2-infected HUVECs with Treg cells ( P<0. 05),but was still higher than that before infection. CYM-5442 pretreatment decreased the expression of IL-6,IL-8 and TNF-α at mRNA level in DENV-2-infected HUVECs and inhibited the secretion of IL-10 and TGF-β by Treg cells that were co-cultured with DENV-2-infected HUVECs. Conclusion Primary HUVECs infected by DENV-2 can enhance the secretion of IL-10 and TGF-β by Treg cells,and the suppressive cytokines produced by Treg cells can reduce the production of inflammatory cytokines by DENV-2-infected HUVECs.

Enforced Expression of CXCR5 Drives T Follicular Regulatory-Like Features in Foxp3⁺ T Cells

CXCR5⁺ T follicular helper (Tfh) cells are associated with aberrant autoantibody production in patients with antibody-mediated autoimmune diseases including lupus. Follicular regulatory T (Tfr) cells expressing CXCR5 and Bcl6 have been recently identified as a specialized subset of Foxp3+ regulatory T (Treg) cells that control germinal center reactions. In this study, we show that retroviral transduction of CXCR5 gene in Foxp3⁺ Treg cells induced a stable expression of functional CXCR5 on their surface. The Cxcr5-transduced Treg cells maintained the expression of Treg cell signature genes and the suppressive activity. The expression of CXCR5 as well as Foxp3 in the transduced Treg cells appeared to be stable in vivo in an adoptive transfer experiment. Moreover, Cxcr5-transduced Treg cells preferentially migrated toward the CXCL13 gradient, leading to an effective suppression of antibody production from B cells stimulated with Tfh cells. Therefore, our results demonstrate that enforced expression of CXCR5 onto Treg cells efficiently induces Tfr cell-like properties, which might be a promising cellular therapeutic approach for the treatment of antibody-mediated autoimmune diseases.

On the correlation between different stages of non-small cell lung cancer with Treg cells and cytokines / 安徽医科大学学报

Objective To investigate the expression of CD4+CDhi25CDlow127 regulatory T cells (Treg cell), IL-2, IL-10 and IFN-γin peripheral blood of patients with different TNM stages of non-small cell lung cancer( NSCLC) , and to evaluate their correlation. Methods The proportion of CD4+CDhi25 CDlow127 Treg cells to CD4+cell in peripheral blood of 55 NSCLC patients were determined by flow cytometry, and levels of IL-2, IL-10 and IFN-γ were measured by ELISA. The proportion of CD4+CDhi25 CDlow127 Treg cells, IL-2, IL-10 and IFN-γ of patients with different TNM stages of NSCLC were compared, and correlation analysis was carried on. Results The proportion of CD4+CDhi25 CDlow127 Treg cells increased obviously together with the increasing of TNM stages of NSCLC(P<0. 05),so did the level of IL-10 (P<0. 05). On the other hand, the level of IL-2 and IFN-γdecreased apparently with the increasing of TNM sta-ges of NSCLC(P<0. 05). CD4+CDhi25CDlow127Treg cells and IL-10 were positively correlated(P <0. 05). IL-2 and IFN-γ were positively correlated too(P<0. 05). CD4+CDhi25CDlow127Treg cells and IL-10 were all negatively correlated with IL-2 and IFN-γ(P<0. 05). Conclusion The increasing of CD4+ CDhi25CDlow127Treg cells and IL-10 and the de-creasing of IL-2 and IFN-γ are possibly one of the reasons of antitumor immunity suppression in terminal NSCLC patients. The surveillance for them can be reference indicators to evaluate antitumor immunity status of terminal lung cancer patients.

The changes of ROR-γt,IL-17A and Foxp3 mRNA expression and promoter methylation in TDI-induced mouse asthma / 中国职业医学

OBJECTIVE: To explore the changes of retinoic acid related orphan receptor-γt( ROR-γt),interleukin( IL)-17 A and forkhead / winged helix protein 3( Foxp3) mRNA expression and promoter methylation in the process of asthma induced by toluene-diisocyanate( TDI). METHODS: Specific pathogens free grade healthy male BALB / c mice were randomly assigned into asthma group and control group with 18 animals in each group. In the asthma group,the mice were sensitized with 0. 30% TDI( mass-volume concentration) dropped on the dorsum of both ears( 20 μL / ear) on day 1 and day 8. On day 15,the mice were challenged with 20 μL 0. 01% TDI( mass-volume concentration) by the trachea. The control group mice were sensitized and challenged by the same procedures with the same amount of solvent( acetone / olive oil). The mice were challenged 24 hours,the pathological changes of trachea and lung tissues were observed. The bronchoalveolar lavage fluid( BALF) from each group was collected,and the inflammatory cells in BALF were counted and classified. IL-4and Interferon-γ( IFN-γ) levels in BALF supernatant were measured by enzyme-linked immunosorbent assay. ROR-γt,IL-17 A and Foxp3 mRNA expression in the lung tissue were measured by real-time fluorescent quantitative polymease chain reaction. The degree of ROR-γt,IL-17 A and Foxp3 promoter methylation in lung issue were detected by Mass Array system.RESULTS: The asthmatic group demonstrated the symptoms of acute asthma,such as breathing deeply and fastly,bowing the back,lifting the forelimbs,et al. But the control group had no such symptoms in mice. Hematoxylin-Eosin staining showed obvious inflammatory lesions in the trachea and lung tissue of asthmatic mice. Compared with the control group,the white blood cell count,the neutrophil and eosinophil percentages in BALF,the IL-4,IFN-γ levels in BALF supernatant in asthma group were all significantly increased( P < 0. 01),meanwhile the lymphocyte and monocyte percentages in BALF were reduced( P < 0. 01); ROR-γt mRNA expression was significantly increased( P < 0. 01),and the degree of promoter methylation from sites 3,4,5,6,8,11 and 12 was significantly reduced( P < 0. 05); IL-17 A mRNA expression was significantly increased( P < 0. 01),and the degree of promoter methylation from sites 6 and 7 was significantly reduced( P < 0. 01); Foxp3 mRNA expression was significantly reduced( P < 0. 01),and the degree of promoter methylation from sites 1 and 10 was significantly increased( P < 0. 01). CONCLUSION: Th17 / Treg cell immune imbalance occurs in asthma induced by TDI. ROR-γt,IL-17 A and Foxp3 gene promoter methylation abnormalities may be involved in Th17 / Treg cell immune imbalance.

The effects of bone marrow mesenchymal stem cells on the treatment of pulmonary fibrosis in silicosis mice / 中国职业医学

OBJECTIVE: To observe the effects of bone marrow mesenchymal stem cells( BMSCs) on the treatment of pulmonary fibrosis in silicosis mice. METHODS: Specific pathogen free healthy male C57BL/6 mice were randomly divided into control group,silicosis group and treatment group with 10 mice in each group. The mice of the control group were given one intra-tracheal injection of 20. 0 μL 0. 90% sodium chloride solution. The silicosis group and treatment group received one 20. 0 μL( mass concentration 250 g/L) of silica dust suspension. After 4 weeks,mice in treatment group were injected with 250. 0 μL of BMSCs suspension( cell density 2 × 10~9/L) by tail vein and silicosis group injected with 250. 0 μL of 0. 90% sodium chloride solution instead,once a week with continuous treatment for 4 weeks. Control group was not given any treatment. Mice were euthanized two weeks after the last treatment. Pathological sections were observed,pulmonary fibrosis score( Ashcroft scores) was marked. Lung coefficient was measured. Lung tissue hydroxyproline( HYP) level and serum transforming growth factor β1( TGF-β1) level were measured. The level of pulmonary fibrosis was scored and the percentages of T helper cell 17( Th17 cell) and regulatory T cell( Treg cell) of spleen and hilar lymph node( HLN) were measured by flow cytometry. RESULTS: The results of lung histopathological examination showed that the pulmonary fibrosis was severe in silicosis group. Massive collagen fiber accumulation and silicotic nodule were found. In treatment group,fibrosis was mild,little collagen fiber accumulation and silicotic nodule were found. The lung coefficient,Aschcroft scores,lung tissue HYP level,serum TGF-β level and the percentage of Th17 cell of spleen and HLN in silicosis group were higher than that of control group( P < 0. 05),while the above indexes of treatment group were lower than that of silicosis group( P < 0. 01). The percentage of Treg cell of spleen and HLN in silicosis group were lower than that of control group( P < 0. 05),while those indexes of treatment group were higher than that of silicosis group( P < 0. 01).CONCLUSION: BMSCs could effectively alleviate the pulmonary fibrosis in silicosis mice and correct the imbalance of Th17/Treg.

Trichomonas vaginalis α-Actinin 2 Modulates Host Immune Responses by Inducing Tolerogenic Dendritic Cells via IL-10 Production from Regulatory T Cells

Trichomonas vaginalis is a pathogen that triggers severe immune responses in hosts. T. vaginalis α-actinin 2, Tvα-actinin 2, has been used to diagnose trichomoniasis. This study was undertaken to examine the role of Tvα-actinin 2 as an antigenic molecule to induce immune responses from humans. Western blot analysis using anti-Tvα-actinin 2 antibodies indicated its presence in the secreted proteins of T. vaginalis. ELISA was employed to measure cytokine production by vaginal epithelial cells, prostate cells, mouse dendritic cells (DCs), or T cells stimulated with T. vaginalis or Tvα-actinin 2 protein. Both T. vaginalis and rTvα-actinin 2 induced cytokine production from epithelial cell lines, including IL-10. Moreover, CD4+CD25− regulatory T cells (Treg cells) incubated with rTvα-actinin 2-treated DCs produced high levels of IL-10. These data indicate that Tvα-actinin 2 modulates immune responses via IL-10 production by Treg cells.

Effects of Hp infection on expression of Foxp3 and RORγt gene in peripheral blood of patients with hepa-titis C cirrhosis / 实用医学杂志

Objective To observe the effect of Hp infection on the expression of Foxp3 and ROR in peripheral blood of patients with hepatitis C cirrhosis ,and to explore the effect of Hp infection on the regulation of Treg cells. Methods Totally 42 hepatitis C cirrhosis patients with Hp infection were chosen as observation group and 30 patients without as control group. Gene expressions were determined by RE-PCR and relative cell factors were detected by ELISA in two groups. Results Expression of FoxP3 and RORγt in observation group was significantly higher than that in control group;the expression level of IL-10,IL-6,IL-17,TGF-β1 and IFN-γin observation group was significantly higher than that in control group but IL-2 expression level was lower than that in control group and expression of IL-10 and TGF-βwas positively correlated with Foxp3 expression and was negatively correlated with IL-2. Conclusion Through inducing the high expression of Treg cells ,Hp may promote the expres-sion of IL-10,TGF-β1,IL-6 and IL-17,inhibit the expression of IL-2 and IFN-γ,imbalance of Treg/Th17 and Th1/Th2 balance in the body and promote the liver inflammation and fibrosis in patients with hepatitis C liver cirrhosis.

Effects of Hp infection on expression of Foxp3 and RORγt gene in peripheral blood of patients with hepa-titis C cirrhosis / 实用医学杂志

Objective To observe the effect of Hp infection on the expression of Foxp3 and ROR in peripheral blood of patients with hepatitis C cirrhosis ,and to explore the effect of Hp infection on the regulation of Treg cells. Methods Totally 42 hepatitis C cirrhosis patients with Hp infection were chosen as observation group and 30 patients without as control group. Gene expressions were determined by RE-PCR and relative cell factors were detected by ELISA in two groups. Results Expression of FoxP3 and RORγt in observation group was significantly higher than that in control group;the expression level of IL-10,IL-6,IL-17,TGF-β1 and IFN-γin observation group was significantly higher than that in control group but IL-2 expression level was lower than that in control group and expression of IL-10 and TGF-βwas positively correlated with Foxp3 expression and was negatively correlated with IL-2. Conclusion Through inducing the high expression of Treg cells ,Hp may promote the expres-sion of IL-10,TGF-β1,IL-6 and IL-17,inhibit the expression of IL-2 and IFN-γ,imbalance of Treg/Th17 and Th1/Th2 balance in the body and promote the liver inflammation and fibrosis in patients with hepatitis C liver cirrhosis.