Effect of Buzhong Yiqitang on Th17/Treg Immune Imbalance and Notch1 Signaling Pathway in AIT Mice / 中国实验方剂学杂志

ObjectiveTo explore the effect of Buzhong Yiqitang on the immune imbalance of helper T cell 17 （Th17）/regulatory T cell （Treg） and Notch1 signaling pathway in mice with autoimmune thyroiditis （AIT）. MethodA total of 60 8-week-old NOD.H-2h4 mice were randomly divided into the normal group， model group， western medicine group （selenium yeast tablet， 32.5 mg·kg-1）， and low-dose （4.78 g·kg-1·d-1）， middle-dose （9.56 g·kg-1·d-1）， and high-dose （19 g·kg-1·d-1） Buzhong Yiqitang groups， with 10 mice in each group. The normal group was fed with distilled water， and the other groups were fed with water containing 0.05% sodium iodide for eight weeks. After the animal model of AIT was formed spontaneously， the mice were killed under anesthesia after intragastric administration for eight weeks. Serum anti-thyroglobulin antibodies （TGAb）， thyroid-stimulating hormone （TSH）， free triiodothyronine （FT3）， and free thyroid hormone （FT4） were detected by enzyme-linked immunosorbent assay （ELISA）， and thyroid tissue changes were observed by hematoxylin-eosin （HE） staining. The mRNA and protein expressions of retinoid-related orphan receptor-γt （RORγt）， interleukin （IL）-17， forkhead box P3 （FoxP3）， IL-10， Notch1， and hair division-related enhancer 1 （Hes1） in thyroid tissue were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultCompared with the normal group， the thyroid structure of the model group was severely damaged， and lymphocytes were infiltrated obviously. The levels of serum TGAb， FT3， and FT4 contents were significantly increased， and TSH content was significantly decreased （P<0.01）. The mRNA and protein expression levels of RORγt， IL-17， Notch1， and Hes1 were significantly increased， while those of FoxP3 and IL10 were significantly decreased in the model group （P<0.01）. Compared with the model group， thyroid structural damage and lymphocyte infiltration were improved in the treatment groups， and serum TGAb， FT3， and FT4 contents were significantly decreased. TSH content was increased， and mRNA and protein expression levels of RORγt， IL-17， Notch1， and Hes1 were decreased. mRNA and protein expression levels of FoxP3 and IL-10 were increased to different degrees （P<0.05， P<0.01）， and the middle-dose Buzhong Yiqitang group had the most significant intervention effect. ConclusionBuzhong Yiqitang can alleviate the thyroid structural damage in AIT mice， and its mechanism may be related to improving the abnormal differentiation of Th17/Treg immune cells and inhibiting the activation of the Notch1 signaling pathway.

Advances in application of regulatory T cells in transplant immune tolerance: from basic to clinical research / 器官移植

Regulatory T cells (Treg) are important inhibitory immune cells to establish immune tolerance, which play a pivotal role in regulating excessive immune response and autoimmune diseases of the host. Previous studies related to transplant immune tolerance have confirmed that increasing the number of Treg in vivo or enhancing the function of Treg serve as a therapeutic strategy to induce transplant immune tolerance. At present, Treg-based induction methods for transplant immune tolerance include adoptive infusion of Treg, in vivo amplification of Treg and utilization of antigen-specific Treg. In this article, the characteristics and mechanism of Treg, the latest research progress on basic experiments and clinical practice of Treg related to transplant immune tolerance at home and abroad were reviewed, and future challenges and development of Treg therapy were prospected, aiming to unravel the significance and application prospect of Treg in transplant immune tolerance, explore the advantages and limitations of Treg therapeutic strategies, and provide reference and evidence for subsequent research in this field.

Immunomodulatory Mechanism of Kangxian Yixin Prescriptions on Autoimmune Injury Mice Based on Th17/Treg Cell Balance / 中国实验方剂学杂志

ObjectiveTo investigate the immunomodulatory mechanism of Kangxian Yixin prescription （KYP） on autoimmune injury mice and its relationship with the T helper 17 （Th17）/regulatory T cell （Treg） balance. MethodSixty healthy 8-week-old male BALBc mice were randomly divided into a normal group and an experimental group at a ratio of 1∶5. On the 0th， 7th， and 28th days， 0.2 mL of porcine cardiac myosin emulsion （containing 0.1 mg of porcine cardiac myosin） was subcutaneously injected into the groin， armpit， and back of the mice in the experimental group to induce an animal model of myocardial immune injury. Mice with myocardial immune injury were randomly divided into a model group （Model）， a KYP group （20.4 g·kg-1·d-1， ig）， and a valsartan group （12 mg·kg-1·d-1， ig）. Mice in the control group and the model group received the same amount of normal saline by gavage. After four weeks of intervention， the heart tissues were collected. Hematoxylin-eosin （HE） staining and Masson staining were used to detect pathological damage in heart tissues. Enzyme-linked immunosorbent assay （ELISA） was used to detect the expression of type B-type natriuretic peptide （BNP）， anti-cardiac antibody， interleukin-17 （IL-17）， and interleukin-10 （IL-10） in the serum of mice， and the expression levels of Th17 cells and Tregs in the spleen were detected by flow cytometry. The protein expression of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X （Bax） in heart tissues was detected by Western blot， and the mRNA expression of retinoid-related orphan receptor gamma t （RORγt） and forkhead box P3 （FoxP3） in the spleen was detected by quantitative real-time polymerase chain reaction （Real-time PCR）. ResultCompared with the control group， the model group showed worsened pathological damage in heart tissues， elevated serum levels of BNP， anti-myocardial antibody， and IL-17， decreased serum expression of IL-10 （P<0.05）， increased expression of Th17 cells and reduced expression of Tregs in spleen tissues （P<0.05）， increased protein expression of Bax， diminished Bcl-2 protein expression， elevated Bax/Bcl-2 ratio， up-regulated mRNA expression of RORγt， dwindled mRNA expression of FoxP3， and elevated ratio of RORγt/FoxP3 （P<0.05）. Compared with the model group， the KYP group and the valsartan group displayed relieved pathological damage in heart tissues， decreased serum expression of BNP， anti-myocardial antibody， and IL-17， increased serum expression of IL-10 （P<0.05）， reduced expression of Th17 cells and increased Tregs in spleen tissues （P<0.05）， dwindled protein expression of Bax and elevated protein expression of Bcl-2 in heart tissues （P<0.05）， diminished Bax/Bcl-2 ratio， reduced mRNA expression of RORγt， up-regulated FoxP3， and down-regulated ratio of RORγt/FoxP3 （P<0.05）. ConclusionKYP may improve myocardial immune damage by regulating the Th17/Treg cell balance.

Regulation Effect and Clinical Efficacy of Addition and Subtraction Therapy of Jinkui Shenqiwan Combined with Buzhong Yiqitang on Immune Inflammatory Factors of Patients with Postmenopausal Osteoporosis and Deficiency of Spleen and Kidney / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of addition and subtraction therapy of Jinkui Shenqiwan combined with Buzhong Yiqitang to postmenopausal osteoporosis （PMO） with deficiency of spleen and kidney， and to investigate its regulation effect on immune inflammatory factors. Method:One hundred and sixty patients were randomly divided into observation group and control group， with 80 cases in each group. Both groups got comprehensive western medicine treatment measures. Patients in control group additionally got Zhuanggu Zhitong capsule， 4 capsules/time， 3 times/day. Patients in observation group additionally got addition and subtraction therapy of Jinkui Shenqiwan combined with Buzhong Yiqitang， 1 dose/day. The treatment was continued for 24 weeks. Before and after treatment， lumbar L2-4 bone mineral density （BMD） was detected by Dual energy X-ray absorptiometry （DXA） and lumbar BMD was detected by quantitative CT （QCT）. Scores of traditional Chinese medicine（TCM） syndromes and Chinese osteoporosis-targeted quality of life questionnaire （COQOL） were graded. Levels of Estradiol （E₂）， type Ⅰ procollagen amino terminal pro peptide （PINP）， serum osteocalcin （OC）， osteoprotegerin （OPG）， type Ⅰ collagen cross-linked C-terminal peptide （S-CTX）， tartrate resistant acid phosphatase （TRACP） and urinary pyridinoline （PYD） were detected. Levels of CD4⁺ T cells， CD8⁺ T cells， interleukin-17 （IL-17）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， <italic>γ-</italic>interferon（IFN-<italic>γ</italic>） and interleukin-4 （IL-4） were calculated. The proportion of T helper cell （Th）17 and regulatory T cell （Treg） in CD4⁺ T cells was calculated. Besides， the safety was evaluated. Result:Bone density was detected by DXA in observation group， and its T-value and bone density detected by QCT were all higher than those in control group （<italic>P</italic><0.01）. After treatment， scores of TCM syndrome and COQOL were lower than those in control group （<italic>P</italic><0.01）. Levels of PINP， OC， S-CTX， TRACP and PYD/Cr were all lower than those in control group （<italic>P</italic><0.01）. Levels of OPG， CD8⁺ and Treg were higher than those in control group （<italic>P</italic><0.05）， levels of Th17， Th17/Treg， CD4⁺/CD8⁺， IL-17， TNF-<italic>α</italic> and IFN-<italic>γ </italic>were lower （<italic>P</italic><0.01）， and levels of IL-4 and E₂ were higher than those in control group （<italic>P</italic><0.01）. The clinical efficacy in observation group was better than that in control group （<italic>Z</italic>=2.103， <italic>P</italic><0.05）. Conclusion:On the basis of calcium and vitamin D supplementation， Jinkui Shenqiwan combined with Buzhong Yiqitang can improve levels of E₂ and bone density， reduce clinical symptoms， improve quality of life， regulate bone metabolism index and immune inflammation reaction， with better clinical efficacy and safety.

Study of the role of CD4+CD45RClowTreg in regulating immune tolerance of rats undergoing liver transplantation / 器官移植

Objective To investigate the role of CD4+CD45RClow regulatory T cell (Treg) in the immune tolerance induction of rats undergoing liver transplantation. Methods Liver transplantation rat models of acute rejection (AR) [Lewis→Brown Norway (BN), AR group] and spontaneous tolerance (BN→Lewis, tolerance group) were established, with 6 rats in each group. Moreover, 3 Lewis rats and 3 BN rats were assigned into the sham operation group (control group). The liver tissues of rats in each group were subject to pathological staining. The expression of T cell subsets and plasmacytoid dendritic cells (pDC) in the peripheral blood, liver graft and spleen of rats was detected in each group. The correlation between pDC and CD4+CD45RClowTreg was analyzed. The expression levels of CD4, CD45RC and CD103 in the liver graft and spleen of rats were quantitatively measured in each group. Results In the AR group, pathological manifestations mainly consisted of inflammatory cell infiltration and structure disorders of transplant liver. Compared with the AR group, the expression levels of CD4+CD25+Treg and CD8+Treg in the peripheral blood were significantly up-regulated in the tolerance group (all P < 0.05). In the peripheral blood, the expression level of CD4+CD25+Treg was positively correlated with that of CD8+Treg (r=0.742, P=0.022). In the AR group, the expression level of CD4+CD45RChighT cell in the peripheral blood was significantly higher than those in the tolerance and control groups (both P < 0.05). Compared with the AR group, the expression level of CD4+CD45RClowTreg in the spleen, and the expression levels of CD8+CD45RClowTreg in the peripheral blood, transplant liver and spleen were significantly up-regulated in the tolerance group (all P < 0.05). Compared with the control and AR groups, the ratio of CD8+CD45RClowTreg/CD8+T in the peripheral blood and the expression levels of pDC in the peripheral blood, transplant liver and spleen were all significantly up-regulated in the tolerance group (all P < 0.05). The expression level of CD4+CD45RClowTreg was positively correlated with the changes of pDC (r=0.506, P=0.016). The expression levels of CD4, CD45RC and CD103 in the transplant liver and spleen of rats were up-regulated in the tolerance group. In the AR group, the expression levels of CD4 and CD45RC were up-regulated, whereas that of CD103 was down-regulated. Conclusions CD4+CD45RClowTreg is a cell subgroup with negative immune regulation, which may construct a regulatory cell network of immune tolerance induction along with CD8+CD45RClowTreg and pDC.

Shaoyaotang Treats Ulcerative Colitis by Inhibiting HIF-1α and Regulating Th17/Treg Balance / 中国实验方剂学杂志

Objective:To explore the effect of hypoxia-inducible factor （HIF）-1<italic>α</italic> on T helper 17 （Th17）/regulatory T cell （Treg） balance in ulcerative colitis and the intervention mechanism of Shaoyaotang. Method:Forty-eight SD rats were randomly divided into normal group （normal saline）， model group， mesalazine group （0.42 g·kg^-1）， Shaoyaotang group （11.1 g·kg^-1）， inhibitor group ［2-methoxyestradiol （2ME₂）， 0.015 g·kg^-1］， and Shaoyaotang+inhibitor group. The ulcerative colitis model was induced by 2，4，6-trinitrobenzene sulfonic acid （TNBS）. The rats in all groups received corresponding treatments for 7 d， and the general condition and disease activity index （DAI） were observed. Hematoxylin-eosin （HE） staining was used to observe histopathological changes of the colon. Enzyme-linked immunosorbent assay （ELISA） was employed to detect serum levels of interleukin （IL）-10， IL-17， and IL-23 in rats. Western blot was used to detect the expression levels of forkhead box protein 3 （FoxP3）， retinoic acid-related orphan receptor <italic>γ</italic>t （ROR<italic>γ</italic>t）， and HIF-1<italic>α</italic> proteins in the colon tissue. Result:Compared with the normal group， the model group showed elevated disease activity index （DAI） score and pathological score for intestinal mucosa （<italic>P</italic><0.01）， reduced serum IL-10 level （<italic>P</italic><0.01）， up-regulated IL-17 and IL-23 levels （<italic>P</italic><0.01）， increased ROR<italic>γ</italic>t and HIF-1<italic>α</italic> expression （<italic>P</italic><0.01）， and decreased FoxP3 protein expression （<italic>P</italic><0.01）. Compared with the model group， the Shaoyaotang group displayed diminished DAI score and pathological score for intestinal mucosa （<italic>P</italic><0.05， <italic>P</italic><0.01）， increased serum IL-10 level （<italic>P</italic><0.01）， decreased IL-17 and IL-23 levels （<italic>P</italic><0.01）， dwindled protein levels of ROR<italic>γ</italic>t and HIF-1<italic>α </italic>（<italic>P</italic><0.01）， and up-regulated expression of FoxP3 （<italic>P</italic><0.01）. Compared with the inhibitor group， the Shaoyaotang group and the Shaoyaotang+inhibitor group exhibited significant differences in the expression of ROR<italic>γ</italic>t， FoxP3， and HIF-1<italic>α</italic> proteins （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:Shaoyaotang could effectively treat ulcerative colitis， and the underlying mechanism of action might be related to the regulation of Th17/Treg rebalance by inhibiting HIF-1<italic>α</italic>.

Traditional Chinese Medicine Intervention on Th17/Treg Balance in Chronic Obstructive Pulmonary Disease： A Review / 中国实验方剂学杂志

Chronic obstructive pulmonary disease （COPD）， a common clinical chronic respiratory disease， has a long course and is intractable. It is closely related to many factors， such as immune imbalance. Helper T cell 17 （Th17）， an immune-promoting cell， and regulatory T cell （Treg）， an immunosuppressive cell， maintain the balance of the immune microenvironment together. In the course of COPD， the proportion of Th17 cells usually increases， while the proportion of Tregs that inhibit Th17 activity decreases. Their coordination and balance are critical in the inflammatory and immune processes of COPD. At present， COPD is mainly treated with nasal inhalation preparations and oral drugs by western medicine. In spite of a certain therapeutic effect， side effects of drugs and heavy economic burden are becoming increasingly prominent. Modern research shows that traditional Chinese medicine （TCM） has the characteristics of few side effects， stable curative effect， and multi-target regulation， and it is advantageous and promising in the prevention and treatment of COPD. In recent years， a large number of TCM clinical and experimental trials on the intervention of Th17/Treg balance in COPD have been launched. Substantial pieces of evidence confirm that the intervention of Th17/Treg balance is an important potential target of TCM in the treatment of COPD. This study reviewed the previous research on the intervention effect of single Chinese medicine， effective components of Chinese medicine， and Chinese medicinal compound on Th17/Treg balance in COPD to comprehensively reveal the potential target of Th17/Treg balance in COPD for clinicians and scientific researchers， promote relevant research， and provide references for the rational application of TCM in the prevention and treatment of COPD.

Application of chimeric antigen receptor-regulatory T cell immunotherapy in organ transplantation / 器官移植

Regulatory T cell (Treg) is a subset of T cells that negatively regulates immunity and has the function of inhibiting rejection. The specific modification of Treg by chimeric antigen receptor (CAR) technology can successfully chime donor-specific antigen onto the surface of Treg, thus regulating the immune function of the body in a real-time manner. It provides a novel and promising therapeutic option for inducing immune tolerance. In this article, research progresses on Treg in immune related diseases, main difficulties in the realization of CAR-Treg technology and its role in inducing transplantation immune tolerance were reviewed, and the opportunities and challenges of CAR-Treg application in the field of organ transplantation are prospected.

The function of regulatory immunological cell in xenotransplantation immunity / 器官移植

Xenotransplantation is the most promising method to resolve the organ shortage problem in the future. In recent years, the advances in gene editing and immunological technique have driven the rapid development of xenotransplantation. However, there are still many insurmountable obstacles in the clinical application of xenotransplantation, among which the rejection is the most important cause of the xenotransplantation failure. Regulatory immunological cells are a group of immunological cells with the negative regulation function in the body, which can inhibit allotransplantation rejection and prolong the survival time of the graft. This paper summarized the research progress of regulatory immunological cells in the xenotransplantation application in recent years, providing reference for the prevention and treatment of xenotransplantation rejection.

Research progress on myeloid-derived suppressor cell and transplantation immune tolerance / 器官移植

Myeloid-derived suppressor cell (MDSC) is a type of heterogeneous cell derived from bone marrow, which was first found in tumor. MDSC can inhibit the function of T cell with immunosuppressive effect. In recent years, more and more studies have shown that in the field of organ transplantation, MDSC can also regulate the host's immune function, induce specific immune tolerance, and play a protective role in transplant organs, which is expected to become a new target in clinical treatment of transplant rejection. The biological characteristics of MDSC and the mechanism of immune tolerance induced by MDSC were reviewed in this paper.

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.

Changes in dendritic cell subsets in mice with type 1 diabetes mellitus / 中华微生物学和免疫学杂志

Objective To investigate the changes in dendritic cell (DC) subsets in mice with type 1 diabetes mellitus (T1DM). Methods A mouse model of diabetes was established through intravenous injection of streptozotocin (STZ) solution. At different time points after STZ treatment, changes in body weight and blood glucose level were observed. Flow cytometry analysis was performed to detect DC subsets, neutrophils, mono-cytes and CD4+Foxp3+regulatory T cells ( Treg) in spleen and lymph nodes. Results STZ treatment led to weight loss and increase in blood glucose in mice. Ten days after STZ treatment, no significant difference in the percentages of DC subsets, neutrophils and monocytes in both spleen and lymph nodes was found between T1DM mice and normal controls. However, spleen volume in the mice treated by STZ significantly reduced, and the percentages of DC subsets in spleen decreased whereas the percentages of neutrophils and monocytes increased as compared with those of normal controls on 30 d and 60 d. Moreover, the percentages of DC subsets, neutro-phils and monocytes in lymph nodes of STZ-treated mice were significantly lower than those of normal controls. Meanwhile, the percentage of CD4+Foxp3+Treg cells in lymph nodes but not in spleen was significantly upregu-lated as compared with that of normal controls. Conclusion Significant changes in DC subsets and other in-flammatory cells in spleen and lymph nodes are observed in mice with STZ-induced diabetes, which reflects the changes in T cell responses and suggests that DC subsets might play an important role in the pathogenesis of T1DM.

Expression and prognostic value of PD-1 and Tregs in metastatic renal cell carcinoma / 中国肿瘤临床

Objectives: To evaluate the tumor-infiltrating PD-1 positive lymphocytes and regulatory T cells (Treg) as prognosis predic-tors of metastatic renal cell carcinoma (mRCC), and investigate the expression of PD-1 and Treg cells in mRCC and elucidate their corre-lation with clinicopathological parameters and prognosis. Methods: A total of 269 mRCC patients from June 2007 to June 2017 in Zhongshan Hospital, Fudan University, were included in the study. The expression of PD-1 and Tregs in mRCC samples were detected by immunohistochemistry. The relationship between the expression of PD-1 and Tregs was analyzed. Results: PD-1 positive expression in mRCC was 31.60% (85/269) and it was positively correlated with the tumor Fuhrman grade and negatively correlated with progno-sis. Tumor infiltration of Tregs in mRCC was 36.80% (99/269), and it was also positively correlated with the tumor Fuhrman grade and negatively correlated with prognosis. Univariate analysis showed that PD-1 positive lymphocytes and high Treg infiltration numbers were negatively correlated with overall survival (OS) and progression free survival (PFS) rates. Thus, PD-1 positive lymphocytes and high Treg infiltration numbers are independent prognostic indicators of OS and PFS and when combined, they can render a better pre-diction for prognosis. Conclusions: Intra-tumoral infiltration of PD-1 positive lymphocytes and Tregs can be used as significant prognos-tic indicators of mRCC, and the combined predictive effect is better than the individual predictive effect. Therefore, evaluating the number of PD-1 positive lymphocytes or infiltrating Tregs in mRCC is helpful in clinically estimating mRCC prognosis in patients.

The function and association between PD-L1 and Treg cells in tumor immunity / 医学研究生学报

Programmed death-1 (PD-1) is the member of the inhibitory costimulatory molecules,has its ligand (programmed death ligand 1,PD-L1),expressing on the surface of a variety of tumor cells.PD-1/PD-L1 plays an important role in the negative regulation of immune responses,promoting tumor immune escape.Regulatory T cell (Treg) is a kind of T cell subsets with immunosuppressive effect,which is highly expressed in tumor tissues,and plays a role in immune tolerance.Recent studies demonstrate key roles of PD-L1 in promoting iTreg cell development and function.This paper gives a brief review of their immune regulations and the association between PD-L1 and Treg.

Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G / 器官移植

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.

The role of the balance between Th17 and Treg in liver disease / 中华肝脏病杂志

Naïve CD4+T cell differentiates into either Th17 or Treg in the microenvironment of various cytokines, among that, Th17 is induced by TGF-β and IL-6, while differentiates into Treg when there is only TGF-β. As the mainly transcription factors of Th17 and Treg respectively, RORγt and Foxp3 have important role in maintaining balance of Th17/Treg. Th17 and Treg are function antagonistically, so the balance of Th17/Treg means a lot for human body. Liver-a metabolic organof the body, it is susceptible to factors in vivo and in vitro. From basal hepatis to end-stage malignancy, common liver diseases are mainly including viral hepatitis, alcoholic liver disease, nonalcoholic fatty liver disease, autoimmune liver disease, hepatocellualr carcinoma and parasitic disease of liver, besides, the occurrence and development of all the diseases are correlated to the balance of Th17/Treg.

Effects of moxibustion on Treg/Th17 cell and its signal pathway in mice with rheumatoid arthritis / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of moxibustion on Treg/Th17 imbalance and related signal pathway in mice with rheumatoid arthritis (RA), so as to explore the action mechanism of moxibustion on RA.</p><p><b>METHODS</b>Twenty-four DBA/1J male mice were randomly divided into a normal group, a model group, a sham moxibustion group and a moxibustion group, 6 mice in each one. RA model was induced by subcutaneous injection of typeⅡcollagen and adjuvant at tail in mice other than the normal group. The mice in the moxibustion group were treated with moxibustion at"Zusanli" (ST 36) and "Shenshu" (BL 23), 1 mg per cone, 6 cones per acupoint. The consecutive 6-day treatment was taken as one course, and totally 2 courses were given with an interval of 2 d between courses. The mice in the sham moxibustion group were treated with immobilization as the moxibustion group. The effects of moxibustion on joint swelling was evaluated by RA scale of collagen induced arthritis (CIA); the pathological changes of joint inflammation were observed by HE staining; the cell count of Th17 and Treg in spleen was analyzed by flow cytometry; the content of cytokine IL-1β, IL-6, IL-10, IL-17, IL-23, TGF-β and Galectin-9 were analyzed by ELISA; the mRNA and protein expression of Foxp3, Galectin-9, RORγt, CARMA1, NF-κB were analyzed by Real-time PCR and Western Blotting method.</p><p><b>RESULTS</b>Ten to 12 d after the secondary immune, red and swelling of ankle joint, feet and toe joint were observed, indicating successful establishment of RA model. 15 d into moxibustion treatment, the joint swelling was improved in the moxibustion group and the sham moxibustion group, which was superior in the moxibustion group (<0.05). As for pathological changes, compare with the normal group, the articular surface was rougher and synovial layer thinner in the model group, which was recovered to a certain extent in the sham moxibustion group; the articular surface was smooth and synovial layer was thicker in the moxibustion group, which was similar to the normal group. The results of flow cytometry test indicated the cell count of Treg in the model group was reduced but that of Th17 was increased than the normal group (both<0.01); the moxibustion could increase significantly the cell count of Treg (<0.05), but no effect was observed on Th17 (>0.05). The results of ELISA test indicated the differences of increasing of IL-1β, IL-6, IL-17, IL-23, TGF-βas well as the reducing of IL-10 were not significant between the sham moxibustion group and the moxibustion group (all>0.05); moxibustion treatment could increase the content of Galectin-9 which was reduced in RA mice (<0.05). The results of RT-PCR and Western blotting test indicated the mRNA and protein expression of Foxp3, Galectin-9 were reduced in the model group (all<0.01), which could be up-regulated by moxibustion treatment (<0.05,<0.01); the mRNA and protein expression of RORγt, CARMA1, NF-κB was increased (all<0.01), which could be down-regulated by moxibustion treatment (<0.05,<0.01).</p><p><b>CONCLUSION</b>Moxibustion could improve the swelling of joint and inflammatory reaction of joint synovial in RA mice; the mechanism may be related to the regulation of Treg cells number in spleen and the expression of Foxp3, Galectin-9, RORγt, CARMA1, NF-κB, mRNA and protein expression.</p>

Effect of azitromycin on Th17/Treg balance in bronchiolitis obliterans mice after lung transplantation / 器官移植

Objective To evaluate the effect of azitromycin upon the bronchiolitis obliterans and T helper (Th)17/regulatory T cell (Treg) balance after lung transplantation. Methods Twenty-four specific pathogen free(SPF) C57BL/6 mice were used as the donors and 48 Balb/c mice were utilized as the recipients. The Balb/c mice were randomly divided into the control (C group), azitromycin control (Cazm group), transplantation (T group) and transplantation + azitromycin groups (Tazm group), 12 mice in each group. In the T and Tazm groups, heterotopic tracheal transplantation models were established to simulate bronchiolitis obliterans after lung transplantation. From 1 d post-transplantation, intragastric administration of azitromycin was given at a dose of 30 mg/kg three times per week in the Cazm and Tazm groups. At 14 and 28 d after transplantation, the transplanted trachea was removed and peripheral blood was collected. The tracheal sample was prepared for hematoxylin-eosin (HE) staining for pathological observation. The expression levels of ROR-γt and Foxp3 messenger ribonucleic acid (mRNA) in the peripheral blood were quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). The variation in the related cytokines levels of Th17 cells and Treg in the plasma was detected by enzyme linked immunosorbent assay (ELISA). Results After heterotopic tracheal transplantation, compared with the C group, thetracheal occlusion accompanied with inflammatory infiltration was observed in the T and Tazm groups. The severity of relevant symptoms in the Tazm group was slighter than that in the T group. Compared with the T group, the expression level of ROR-γt mRNA in the Tazm group was significantly down-regulated (P<0.05). No statistical significance was identified in the expression of Foxp3 mRNA between two groups (P>0.05). Compared with the T group, the levels of interleukin (IL)-6 and IL-17 cytokines in the Tazm group were significantly down-regulated (all P<0.05). Conclusions Persistent therapy of azitromycin can delay the progression of bronchiolitis obliterans after transplantation, which is probably associated with inhibiting Th17 cell differentiation and inflammation.

Roles of regulatory T cell in tumor immunity and tumor immunotherapy / 中国肿瘤生物治疗杂志

There are multiple types of inhibitory immune cells in tumor. Among these cells, Treg (regulatory T cell) plays an extremely important role in tumor development and progression. Treg exihibits potent inhibitory effects on effector cells by a variety of mechanisms, which might be the the key factor for tumor immune escape. These mechanisms include inhibiting the effector cell function by inhibitory cytokines, killing effector cells by granzyme and profrin, interfering effector cell metabolism, and affecting Treg differentiation and proliferation by regulating the function of dendrtic cells, etc. The research on Treg has provided new strategies for tumor immunotherapy. Tumor immunotherapies targeting Treg and related immunosuppressive factors, such as deleting Treg nonsepcificlly or sepcificlly controling the numbers and functions of Treg, might have a bright future in clinical application.keyword regulatory T cell(Treg); neoplasms; immune escape; immunotherapy