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Dioscin has been used as a treatment for Hashimoto’s thyroiditis (HT) in China. However, the molecular mechanisms governing the modes of action of dioscin have not been elucidated. In this study, f...

Aims. To determine the role of regulatory cells (Treg) in patients with vulgar psoriasis (VP) and to provide evidence of its possible role in diagnosis, treatment, and measurement of therapeutic efficacy. Material and methods. We studied 60 patients (35 women and 25 men) with VP, ages ranging from 18 to 55 years. Of these, 28 patients had VP in the progressive stage, 19 were stable, and 8 had resolution of the disease. Overall, 42 patients had VP for less than 20 years and 28 patients for 20 years and more. Patients were divided based on VP severity into the following three groups: mild (10 patients), moderate (22 patients), and severe (28 patients). In addition, the amount of Treg was examined before and after narrow-band ultraviolet-B (UVB) therapy in 12 patients with the progressive stage of VP. The healthy donors (HD) group consisted of 22 persons. Results. A reduction in the number of Treg, CD4+CD25+Foxр3+CD127LOW, was observed in the peripheral blood of patients with VP (2.84% 1.00% for patients with VP and 4.02% 0.73% for HD), with an increase in their number with stage transition (2.59% 0.68% in the progressive stage, 2.82% 1.55% in stable, and 3.68% 1.62% in the resolution period). An inverse correlation was determined not only between the number of Treg and the degree of VP severity (r = -0.39) but also with disease duration (r = -0.46). In addition, NB-UVB phototherapy was noted to promote an increase in the amount of Treg.

Background: CXCL4 plays an essential role in the regulation of multiple immune diseases. However, the underlying role of CXCL4 is still not clear in sepsis. Aim: In the present study, we aimed to i...

Aim: The value of the peripheral blood lymphocyte subpopulation ratios and tumor diameter for prognosis in bladder cancer (BC) patients needs to be explored. Materials & methods: A total of 161 male BC patients and 68 male normal controls were retrospectively reviewed. The value of combining predictor consisted of both CD4+CD25+/CD4+ and computed tomography urography tumor diameter (CTU-D) on stage, overall survival (OS) and recurrence probability was analyzed by logistic regression, Kaplan–Meier method and log-rank test. Results: The combining predictor was a statistically independent risk for stage; dramatic differences in OS and recurrence probability were found between the combining predictor-high (cut-off point >0.08) and combining predictor-low groups (cut-off point ≤0.08). Conclusion: The combining predictor could be a significant predictor for advanced stage, OS and recurrence probability in male patients with BC.

Attention deficit hyperactivity disorder (ADHD) is a most common mental illness in both children and adults. Our recent studies revealed that high-dose taurine improves hyperactivity in SHR rats by reducing mALFF signal and striatal dopamine uptake. This study further revealed the association between immune factors and hyperactivity in SHR rats fed with high-dose taurine. A positive correlation was detected between systolic blood pressure (SBP) and horizontal locomotion in SHR rats fed with high-dose taurine. Significantly higher striatal Hsp27 and galectin-3 were detected in SHR rats fed with high-dose taurine. Significantly lower IL-2 and IL-6 were detected in SHR rats fed with high-dose taurine, whereas significantly higher IL-10 was detected. Significantly increased splenic CD4+CD25+Foxp3+ regulatory T cells was detected in SHR rats fed with high-dose taurine with a negative correlation. These findings suggest that high-dose taurine reduce hyperactive behavior in SHR rats probably via multifactorial modulation on immune system.

Background: Among CD4+ T helper (Th) cells, Th17-Treg imbalance is a major pathogenesis of Chronic periodontitis (CP). Treg cells are often considered to play a protective role but they may have plasticity. This study aimed to clarify the regulatory role of Th17-Treg balance in periodontitis and further reveal Treg plasticity.Methods: An experimental periodontitis model was established by ligation of a silk thread and local injection of Pg-LPS. Inflammatory factors in serum and gingival tissues were measured by ELISA and RT-PCR. The degree of alveolar bone absorption was evaluated by micro-CT and histomorphological analysis. Quantities of Treg and Th17 cell and their related gene expression levels were examined. Furthermore, after magnetic bead-sorting spleen Treg cells, their characteristic genes and Th17 cell-related factors were explored at the mRNA level.Results: Inflammatory cytokines in serum and gingival tissue increased significantly in experimental periodontitis, which revealed obvious crestal bone loss around maxillary second molars. Consistently, we found increased secretion of RANKL by osteoblasts, thereby promoting the formation of osteoclasts and enhancing their activity in periodontitis. Further analysis showed that the number of Th17 cells and expression of related genes increased more significantly than Treg cells, demonstrating Treg-Th17 imbalance in periodontitis. Flow cytometry showed that the proportions of Treg cells in the blood and spleen of the periodontitis group was lower than those of the control group. Furthermore, Treg cell-related gene Foxp3 was downregulated and their expression of Th17 cell-related genes Rorc and IL-17A were increased. Conclusions: These results provided evidence that periodontitis may lead to Treg-Th17 conversion, although its mechanisms requires further study.

Background: Several clinical trials on cancer showed a correlation between elevated levels of regulatoryT cells with either poor prognosis or poor response to some therapies. Hence, in this study we tried tomeasure the regulatory T cells (CD4+CD245+) count and to evaluate the program death receptor 1(PD1)as a one of the main suppressive mechanisms that regulatory T cells use in CML patients with differentmolecular response to imatinib therapy. Method: Flow cytometry technique was used to analyze 30 sampleof optimal molecular response of CML patients (BCR-ABL transcripts ? 0.1%) and 30 sample of failuremolecular response patients (BCR-ABL transcripts >1%) with or without hematological failure, in order toassess the CD4+CD25+ T cells (Tregs) and PD1 expression on these cells. Thirty samples age and gendermatched were used as healthy controls. Results: A high proportion of CD4+CD25+ T cells was found infailure groups compared to control and optimal groups(P= 33.7 % washighly significant with high sensitivity and specificity. This value can be used to determine the failure fromthe optimal CML responders. There was no noticeable differences (P= 0.125) in PD1+ expression amongCD4+CD25+ T cells. Conclusion: A high percentage of Treg cells in the failure CML group might be anindication of immune escape of leukemic cells in those patients compared to the other investigated groups.

Thymic stromal lymphopoietin (TSLP) is a cytokine and is closely related to Interleukin (IL) - 7, and hTSLP can activation through the human thymus dendritic cell in thymic to indirectly promote the differentiation of natural Regulatory T cells (Tregs) of the thymus. In this study, we focused on recombinant TSLP to determine its effects on the differentiation of CD4+CD25-T cells separated from the thymus of myasthenia gravis (MG) patients. Our results demonstrated that exogenous TSLP could increase CD4+CD25+T/CD4+T cells ratio, up-regulate the expression of Foxp3 mRNA and protein expression in CD4+CD25+Treg cells. Furthermore, we found that CD4+CD25+ Treg cells induced by exogenous TSLP could secrete IL - 10, Transforming growth factor (TGF) - β and the ability to inhibit CD4+T cell proliferation improved. These results indicate that TSLP may promote the differentiation of thymic CD4+CD25-T cells of MG patient to CD4+CD25+Foxp3+ regulatory T cells and enhance the function of immune suppression.

This review focuses on the role of regulatory T cells (Tregs) in the process of carcinogenesis. The controversy of this issue arose due to the increasing therapeutic use of Tregs in humans (inter alia, in the treatment of autoimmune diseases). It is mainly due to potential dangers related to immunosuppressive activity of these cells, especially regarding cancer. The natural function of regulatory T cells (which is the suppression of excessive activity of the immune system) is purportedly linked to an increased risk of cancer initiation. This work brings together and summarizes the most important reports of researchers dealing with this problem and attempts to explain doubts and fears related to Tregs and their uncertain connection with cancer initiation and progression. It is clearly shown that regulatory T cells are associated with acceleration of existing tumors (they are attracted by microenvironments created by cancer cells) but cannot initiate them on their own.

Background Systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) as autoimmune diseases arise owing to failure of immunological self-tolerance. One of the mechanisms employed to control these potentially damaging cells are regulatory T cells (Tregs). The importance of Tregs is underscored by the overwhelming inflammation and autoimmunity that result from their absence. Forkhead box p3 (FoxP3) is an important regulator of Treg function, and the expression of FoxP3 correlates with the expression of other Treg-associated markers such as CD25 and CTLA-4. Aim To investigate the frequency of FoxP3+ CD4+ CD25+high cells (Tregs) in peripheral blood from patients with SLE and those with RA. Patients and methods A total of 25 patients with SLE (15 patients with active SLE and 10 patients with inactive SLE), 25 patients with RA (15 patients with active RA and 10 patients with inactive RA), and 10 age-matched and sex-matched healthy controls were enrolled in the study. Patients underwent clinical and laboratory assessment. The frequency of Tregs was determined by flow cytometry. Results The distribution of FoxP3+ CD4+ CD25+high cells (Tregs) revealed a highly significant decrease in the frequency of Treg in patients with SLE compared with healthy controls. Moreover, patients with active SLE showed significantly lower Tregs percent when compared with inactive group. Moreover, the distribution of FoxP3+ CD4+ CD25+high cells (Tregs) revealed a high significantly decrease in the frequency of Treg in patients with RA compared with healthy controls. Conclusion CD4+ CD25+ FoxP3 Tregs (as a percent of total CD4 cells) were significantly lower in patients with SLE and those with RA when compared with healthy controls.

Multipl Skleroz patogenezinde otoimmunitenin ortaya cikisinda periferik immun toleransin bozulmasi onemli rol oynamaktadir. Immun toleransin saglikli isleyisinde regulatuvar T hucreleri ise anahtar rol oynamaktadir. FoxP3 ise regulatuvar T hucreler yeterli ekpresyonu icin transkripsiyon faktoru olarak gorev almaktadir. Biz bu calismayla Mc Donald’s kriterlerine gore klinik olarak kesin MS tanisi almis 12 RRMS (relapsing Remitting Multipl skleroz), 11 SRMS ( Sekonder Relapsing Multipl skleroz), 8 atak RRMS hastasi olmak uzere toplam 31 hasta ile 12 saglikli kontrol gurubunun regulatuvar T hucre alt tipleri hematoloji laboratuarinda akim sitometrisi ile analiz ederek total CD4(+) T lenfosit icerisindeki yuzdeleri ve FoxP3 ekspresyonunu karsilastirdik. Atakli 8 RRMS hastasinin ise 7 tanesi 1000 mg/ gun IVMP tedavisinden once ve tedaviden sonra kendi icinde karsilastirildi. Sonuclar istatiksel olarak degerlendirildi. Regulatuvar T hucre profili CD4(+) CD25(+), CD4(+) foxP3(+), CD4(+) CD25(+) foxP3(+),CD4(+) CD25(+) foxP3(-), ve CD4(+) CD25(-) foxP3(+) karsilastirildi, hastalar ve kontrol grubu arasinda anlamli fark bulunmadi (p>0.005). Atak oncesi ve atak sonrasi IVMP tedavisi sonrasi 7 hastanin regulator hucre profilleri ve foxP3 ekspresyonu arasinda anlamli fark saptanmadi (p>0.05). Ancak, yuksek doz intravenoz metilprednizolon (IVMP) tedavisinin regulator hucre alt tipleri icinde hafif bir sayisal artis neden oldugu gozlenmistir. Regulatuvar T hucrelerin MS imunopatogenezinde sayisal yeterliligi yaninda islevsel ozelliklerinin onemli rol oynadigi dusunuldu.

Objective Early detection of severe forms with unfavorable outcome is the cornerstone that could provide reduction of morbidity and mortality in acute pancreatitis (AP). Methods The percentage of circulating CD4CD25CD127 regulatory T-cells (Tregs) was determined at admission, on the 48th hour, and on the fifth day in 72 patients with AP. We divided patients in 2 groups-Sev1, which includes 19 patients (26.4%) with moderate AP and 39 patients (54.2%) with mild disease, and Sev2, which includes 14 patients (19.4%) with severe AP. Seven patients (9.7%) developed septic complications. The mortality in our group was 9.7%. Results The patients in Sev2 had higher percentage of Tregs at admission and on the fifth day compared with patients in Sev1 (P = 0.007 and P = 0.033, respectively). There was no significant difference in percentage of Tregs at admission, on the 48th hour, and on the fifth day in patients who developed and did not develop infected necrosis (P = 0.50, P = 0.72, and P = 0.92, respectively). Patients with poor outcome had elevated percentage of Tregs on the fifth day (P = 0.045). Conclusions The percentage of circulating Tregs may be implicated in the development of early immune suppression in AP. Elevated percentage of circulating Tregs at admission in AP is an independent prognostic biomarker for severe disease.

Aim To investigate T-cell subpopulations in peripheral blood of human parvovirus B19 DNA-positive (B19+) and -negative (B19-) patients with rheumatoid arthritis (RA) and healthy persons. Patients and methods Blood samples were collected from 115 patients with RA and 47 healthy volunteers; 27 patients with RA and nine controls were B19+ Cluster of differentiation (CD) 4, 8, 25 and 45RA were analyzed on blood cells. CD25 expression on CD4+CD45RA+, CD4+CD45RA-, CD8+CD45RA+, CD8+CD45RA- subsets were analyzed by flow cytometry. Results The percentage of CD25low and CD25hi cells was increased on CD4+CD45RA+, CD4+CD45RA- T-cells and the percentage of CD25+ cells was increased on CD8+CD45RA+, CD8+CD45RA- T-cells of B19+ patients with RA in comparison with B19- patients and controls. Conclusion Raised levels of CD4 and CD8 regulatory T-cells in B19+ RA patients could cause down-regulation of antiviral clearance mechanisms and lead to activation of persistent human parvovirus B19 infection in patients with RA.

Background Regulatory T cells (Tregs) play a crucial role in maternal-fetal tolerance, but little is known about the characteristic of Tregs in peripheral blood (PB), maternal-fetal interface and cord blood (CB) in normal and complicated pregnancies with preeclampsia (PE), gestational diabetes mellitus (GDM) and premature rupture of membranes (PROM). Methods PB, retro-placental blood (RPB), and CB were collected immediately after delivery in women with normal full-term pregnancy (NP), PE, GDM, and PROM. The proportion of CD4 + CD25 + CD127 low/- T cells (Tregs) and the expression of PD-1, GITR, HLA-G and CTLA-4 on T cell subsets were investigated by flow cytometric analysis. The data were analyzed based on sample origins (PB, RPB, and CB) and the obstetrical study groups (NP, PE, GDM, and PROM). Results The proportions of Tregs in PB, RPB, and CB from NP were significantly higher than the other obstetrical groups (P < 0.01, respectively). However no significant differences were present among the PE, GDM and PROM groups (P = NS). The proportion of PD-1 + and GITR + Tregs in RPB and PB as well as PD-1 + Tregs in CB from NP were significantly higher than those of the same origin in each obstetrical study group (P < 0.01, respectively). There were no differences in HLA-G + and CTLA-4 + Tregs between different origins in each obstetrical study group. In NP, the proportion of PD-1 + Tregs was significantly decreased in CB as compared to those of PB and RPB (P < 0.05, respectively). Among the four groups, the proportion of GITR + Tregs were significantly higher in PB as compared to those of CB and RPB (P < 0.01, respectively). The proportion of HLA-G + Tregs in PB was significantly lower than that of CB and RPB (P < 0.01, respectively). The proportion of CTLA-4 + Tregs had no significant differences (P = NS). Conclusions In conclusion, the proportion and characteristics of Tregs vary in the maternal, fetal and maternal-fetal junction at the time of delivery. The different feature and density of Tregs in maternal and fetal tissue may suggest the multiple and complicated roles of Tregs during pregnancy.

Multiple sclerosis (MS) is an autoimmune disease characterized by defect in regulatory function of CD4+CD25+ T cells. We demonstrated difference in proportion of regulatory T cells CD4+CD25+FoxP3+CD127low (Tregs) within the same patients’ relapse and remission. Proportion of peripheral Tregs (pTregs) dropped almost two times in the relapse compare to remission. Levels of pTregs in patients’ remission were lower than in healthy donors. Suppressive ability of pTregs was decreased in MS patients compared to healthy donors. Injections of expanded ex vivo autologous Tregs (eTregs) could be helpful in bringing up the level of Tregs in patients’ blood. We developed a simple method for ex vivo expansion of autologous Tregs within a short period of time. The final pool of cells consisted of 90-95% eTregs. When we started the culture with 10-20 × 106 CD4+ T cells, we yield 300-400 × 106 eTregs in a week. Expression of FoxP3 and Helios was calculated by two methods. Expanded ex vivo patients’ and donors’ Tre...

Background It was found that regulatory T cells (Tregs) importantly affect the maintenance of the kidney graft. However, Tregs are a heterogeneous population with less to more suppressive activity. The aim of this study was to determine the effects of different subsets of Tregs, as well as their ratio to effector T cells (Teff), on kidney transplantation outcomes. Methods A total of 58 participants were enrolled in this study and divided into four groups: (i) first kidney transplant recipients (stable 1); (ii) second kidney transplant recipients (stable 2); (iii) transplant recipients with acute rejection (AR); and (iv) healthy control subjects. By using flow cytometer, the frequencies of CD4+ CD25++ CD45RA− Foxp3hi activated Tregs (aTregs), CD4+ CD25+ CD45RA+ Foxp3lo resting Tregs (rTregs), CD4+ CD25+ CD45RA− Foxp3lo non-suppressive T cells, CD4+ CD25+ Foxp3− cells Teff, and total Tregs were analyzed in all subjects. Results The frequency of aTregs (as well as the ratio of aTregs/Tregs) was significantly lower in the AR patients than the other three groups. In contrast to AR patients, stables 1 and 2 had a higher aTreg/Treg ratio than those in the control group. Although patients with AR had a significantly lower total Tregs than the other three groups, the balance of total Tregs and Teff was similar between patients with and without AR. Conclusion Patients with AR had poorer immunoregulatory properties than those with normal graft functioning, as well as those in the control group. These reduced immunoregulatory properties in patients with AR could lead to graft rejection.

Effects of anesthesia methods on immune function in patients with lung cancer undergoing radical operation were investigated. A total of 122 patients undergoing radical resection of lung cancer who were treated in Zhejiang Cancer Hospital from September 2013 to April 2016 were randomly divided into the combined anesthesia group and the intravenous anesthesia group, with 61 cases in each group. The patients in the combined anesthesia group were given intravenous combined epidural anesthesia. Patients in the intravenous anesthesia group were given intravenous anesthesia. The change of CD3+, CD4+ and CD4+CD25+ at time-point T0 (before anesthesia), T1 (the time of anesthesia), T2 (after operation), T3 (24 h after operation), T4 (72 h after operation) were compared between the two groups. The levels of CD3+, CD4+ and CD4+CD25+ at T1, T2, T3 and T4 in the combined anesthesia group were higher than that in the intravenous anesthesia group (P

A rapidly growing evidence showed that regulatory T cells (Tregs) play a crucial role in tolerance to coagulation factors and may be involved in the pathogenesis of inhibitor formation in patients with hemophilia. We determined the percentage of Tregs (CD4CD25CD127) in 45 children with hemophilia A compared with 45 healthy controls, and assessed their relation to the clinical characteristics of patients and factor VIII (FVIII) inhibitors. Patients were studied stressing on frequency of bleeding attacks, joint pain, history of viral hepatitis, and the received therapy (FVIII precipitate/cryotherapy). FVIII activity and FVIII inhibitors were assessed with flow cytometric analysis of CD4CD25CD127 Tregs. According to residual FVIII activity levels, 30 patients (66.7%) had mild/moderate hemophilia A, whereas 15 (33.3%) patients had severe hemophilia A. The frequency of Tregs was significantly lower among all patients with hemophilia A compared with controls (2.59 ± 1.1 versus 3.73 ± 1.12%; P = 0.002). Tregs were significantly decreased among patients with FVIII inhibitors compared with the inhibitor-negative group (P < 0.001). Patients with hematuria or severe hemophilia A had lower Tregs levels than those without (P = 0.34 and P = 0.011, respectively). A significant positive correlation was found between the percentage of Tregs and FVIII among hemophilia A patients. ROC curve analysis revealed that the cut-off value of Tregs at 1.91% could differentiate patients with and without FVIII inhibitors, with a sensitivity of 100% and a specificity of 91.3%. We suggest that alteration in the frequency of Tregs in young patients with hemophilia A may contribute to inhibitor formation and disease severity.

Cystic fibrosis (CF) lung disease is characterized by dysregulated inflammatory response in the airways. CD4(+)CD25(+) regulatory T cells play a crucial role in maintaining the immune homeostasis. However their role in the disease pathogenesis of CF remains unexplored.To determine the percentage of circulating CD4(+)CD25(high), FoxP3(+) T cell expression in children with CF and controls. Furthermore to evaluate the relationship between CD4(+)CD25(high), FOXP3 T cell % and the clinical status of the disease (lung function).CD4(+)CD25(+), intracellular FoxP3 expression in peripheral blood were estimated using flowcytometry in 20 children with CF and 10 healthy controls. Spirometry was carried out according to the standard guidelines.We observed a significant difference in CD4(+)CD25(+)T cell% in children with CF (5.2 ± 1.2) versus controls (6.8 ± 1.4, p0.05), CD4(+)CD25(high)T cell% in CF (1.72 ± 0.36) versus controls (2.59 + 0.42, p0.003). Similarly a significant difference was observed in FoxP3 T cell% CF: (60.7 ± 6.19) versus controls (76.8 ± 5.16), p0.001. A significant positive correlation between FoxP3 T cell% and FEV1 in children with CF(r = 0.822, p0.01) was observed.CD4(+)CD25(high) T cell% correlated positively with FEV1 (r = 0.742, p0.01).Our findings report the first evidence of a decreased expression of circulating CD4(+)CD25(high) FoxP3(+) T cells in children with CF. Furthermore circulating CD4+ CD25(high), FOXP3(+) T cell percentage correlated with FEV1.

Background/Purpose(s)We investigated whether CD4+CD25+Foxp3+ regulatory T cells (Tregs) are induced in patients suffering from early-stage septic shock and distinguish them from noninfectious patients with systemic inflammatory response.MethodsThe study included 37 patients with early-stage septic shock, 15 patients with noninfectious systemic inflammatory response syndrome (SIRS), and 24 heath controls. We prospectively assayed the fraction of Tregs expressing high levels of CD25 and forkhead box P3 (Foxp3) as well as the plasma levels of interferon-γ (IFN-γ), interleukin-4 (IL-4), and soluble CD25 in all the subjects studied.ResultsCompared with the control groups, the plasma levels of IFN-γ [66.10 (45.23–85.08) pg/mL vs. 20.97 (17.58–26.21) pg/mL, p

To investigate the expression of CD4+CD25+T cells and the Foxp3 in peripheral blood of rheumatoid arthritis（RA），and to analyze the relationship between their activities and patho- genesis.The number of CD4+CD25+T lymphocytes in the peripheral blood and Foxp3 mRNA expression on peripheral blood monocytes of 48 RA patients and 35 normal controls were analyzed by three-color FACs analysis and reverse transcription-polymerase chain reaction（RT- PCR）．The expression of CD4+CD25+T cells RA patients in active group was significantly lower than that in remission group and healthy controls（P0.05）； The relationship between peripheral blood CD4+CD25+Tregcells as well as the Foxp3 mRNA and active renal score was negatively correlated. The expression of CD4+CD25+Tregcells and the Foxp3 mRNA of peripheral blood in RA patients is abnormal and it is correlated with pathogenesis and activity of RA.

Jingya Li,1,* Ruizhen Zhao,1,* Xiaoli Li,1 Wenjun Sun,1 Miao Qu,1 Qisheng Tang,1 Xinke Yang,1 Shujing Zhang2 1Third Affiliated Hospital, 2School of Basic Medical Sciences, Beijing University of Chinese Medicine, Beijing, People’s Republic of China *These authors contributed equally tothis work Background: Shen-Qi-Jie-Yu-Fang (SJF) is composed of eight Chinese medicinal herbs. It is widely used in traditional Chinese medicine for treating postpartum depression (PPD). Previous studies have shown that SJF treats PPD through the neuroendocrine mechanism. Aim: To further investigate the effect of SJF on the immune system, including the inflammatory response system and CD4+CD25+ regulatory T (Treg) cells. Materials and methods: Sprague Dawley rats were used to create an animal model of PPD by inducing hormone-simulated pregnancy followed by hormone withdrawal. After hormone withdrawal, the PPD rats were treated with SJF or fluoxetine for 1, 2, and 4 weeks. Levels of Treg cells in peripheral blood were measured by flow cytometry analysis. Serum interleukin (IL)-1β and IL-6 were evaluated by enzyme-linked immunosorbent assay, and gene and protein expressions of IL-1RI, IL-6Rα, and gp130 in the hippocampus were observed by reverse-transcription polymerase chain reaction and Western blot. Results: Serum IL-1β in PPD rats increased at 2 weeks and declined from then on, while serum IL-6 increased at 1, 2, and 4 weeks. Both IL-1β and IL-6 were downregulated by SJF and fluoxetine. Changes in gene and protein expressions of IL-1RI and gp130 in PPD rats were consistent with changes in serum IL-1β, and were able to be regulated by SJF and fluoxetine. The levels of Treg cells were negatively correlated with serum IL-1β and IL-6, and were decreased in PPD rats. The levels of Treg cells were increased by SJF and fluoxetine. Conclusion: Dysfunction of proinflammatory cytokines and Tregs in different stages of PPD was attenuated by SJF and fluoxetine through the modulation of serum concentrations of IL-1β and IL-6, expressions of IL-1RI, and gp130 in the hippocampus, and CD4+CD25+ Treg cells in peripheral blood. Keywords: postpartum depression, inflammatory cytokines, antidepressant drugs, Chinese herbal medicine, shenqi jieyu fang, animal model

Myasthenia gravis (MG) is a CD4(+) T cell-dependent autoimmune disease, and close attention has been paid to the role of CD4(+)CD25(+)Treg cells (Tregs). Previous results regarding Tregs in MG patients have been conflicting. The discrepancy was partly ascribed to selecting different Treg-associated molecules in defining Tregs. Therefore, we considered it necessary to find a reliable index for assessing the immunologic state in MG patients and explore the effect of IS on them.We adopted flow cytometric techniques to measure the numbers and frequencies of Tregs in peripheral blood taken from 57 patients and 91 age-matched healthy donors, and we also analyzed FOXP3 mean fluorescence intensity on Tregs.The number and frequency of Tregs in peripheral blood of MG patients significantly decreased, together with down-regulation of FOXP3 expression. There was dynamic change of Treg cell level and the inverse relationship with clinical symptom, suggesting that the immunologic disorder in MG patients was related to peripheral Tregs population. Meanwhile, CD4(+)CD25(+)FOXP3(+)Helios(+)T cells might be activated Tregs, rather than nTregs. Moreover, the number and frequency of CD4(+)CD25(+)FOXP3(+)Helios(+)T cells significantly decreased in MG patients, indicating that the reduction of the activated Tregs population might be a critical contributor to the pathogenesis of MG.The significant reduction of the peripheral Tregs population in MG patients might be responsible for the immunologic disorders in MG patients. IS such as GC took its effect possible by increasing the population size, and the underlying mechanism should be further investigated.

Inflammatory bowel disease (IBD), which is characterized by a dysregulated intestinal immune response, is postulated to be controlled by intestinal self-antigens and bacterial Ags. Fecal extracts called cecal bacterial Ag (CBA) have been implicated in the pathogenesis of IBD. In this study, we identified a major protein of CBA related to the pathogenesis of IBD and established a therapeutic approach using Ag-pulsed regulatory dendritic cells (Reg-DCs). Using two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry, carbonic anhydrase I (CA I) was identified as a major protein of CBA. Next, we induced colitis by transfer of CD4+CD25− T cells obtained from BALB/c mice into SCID mice. Mice were treated with CBA- or CA I-pulsed Reg-DCs (Reg-DCsCBA or Reg-DCsCA1), which expressed CD200 receptor 3 and produced high levels of IL-10. Treatment with Reg-DCsCBA and Reg-DCsCA1 ameliorated colitis. This effect was shown to be Ag-specific based on no clinical response of irrelevant Ag (keyhole limpet hemocyanin)-pulsed Reg-DCs. Foxp3 mRNA expression was higher but RORγt mRNA expression was lower in the mesenteric lymph nodes (MLNs) of the Reg-DCsCA1–treated mice compared with those in the MLNs of control mice. In the MLNs, Reg-DCsCA1–treated mice had higher mRNA expression of IL-10 and TGF-β1 and lower IL-17 mRNA expression and protein production compared with those of control mice. In addition, Reg-DCsCBA–treated mice had higher Foxp3+CD4+CD25+ and IL-10–producing regulatory T cell frequencies in MLNs. In conclusion, Reg-DCsCA1 protected progression of colitis induced by CD4+CD25− T cell transfer in an Ag-specific manner by inducing the differentiation of regulatory T cells.

The immunosuppressive properties of regulatory T cells have emerged as an attractive tool for the development of immunotherapies in various disease contexts, e.g. to treat transplantation induced immune reactions. This paper focuses on the process of obtaining and functionally characterizing CD4+CD25+FoxP3+ regulatory T cells and Tr1 cells from uremic patients awaiting kidney transplantation.From October 2010 to March 2011 uremic patients awaiting living donor kidney transplantation, and their corresponding kidney donors, were enrolled in the study. A total of seven pairs were included. Isolation of CD4+CD25+FoxP3+ regulatory T cells was performed by magnetic activated cell sorting of peripheral blood mononuclear cells obtained from the uremic patients. Donor specific Tr1 cells were differentiated by repetitive stimulation of immature CD4+ T cells with immature dendritic cells, with the T cells coming from the future kidney recipients and the dendritic cells from the corresponding kidney donors. Cells were then expanded and functionally characterized by the one-way mixed leukocyte reaction and assessment of IL-10 production. Phenotypic analysis was performed by flow cytometry.The fraction of CD4+CD25+FoxP3+ regulatory T cells after expansion varied from 39.1 to 50.4% and the cells retained their ability to substantially suppress the mixed leukocyte reaction in all but one patient (3.8-19.2% of the baseline stimulated leukocyte activity, p0.05). Tr1 cells were successfully differentiated from all but one patient and produced high levels of IL-10 when stimulated with immature dendritic cells (1,275-11,038% of the baseline IL-10 secretion, p0.05).It is practically feasible to obtain and subsequently expand CD4+CD25+FoxP3+ regulatory T cells and Tr1 cells from uremic patients without loss of function as assessed by in vitro analyses. This forms a base for adoptive regulatory T cell therapy in the setting of living donor kidney transplantation.

CD4 + CD25 + regulatory T-cells: a potential target for treating patients with chronic HBV infection

Background: CD4+CD25+ regulatory T (Treg) cells can control the allergic response to allergen, airway eosinophilia and airway hypersensitivity. We speculated that chronic inflammation persisting in asthma airways is dependent on abnormalities of these Treg cells. There are differences in the pathology of asthma in adults and children, and the airways of pediatric asthma are considered to be more naive than those of adults. Therefore, we analyzed the functionality of Treg cells in pediatric asthma and the relationship between Treg function and asthma symptoms. Methods: The anergic state, which is one of the defining properties of Treg, was analyzed by measuring intracellular Ca2+ influx following T cell receptor (TCR) stimulation. FOXP3-positive cells and FOXP3 mRNA expression were measured by flow analysis and real-time PCR with the SYBR method, respectively. Results: CD45RO+ cells make up approximately 99% of CD4+CD25high T cells and 89% of CD4+CD25low T cells in human adult blood. The proportion of CD45RO+ cells in CD4+CD25+ (high + low) T cells from pediatric asthma was much smaller (about 56%). Interestingly, our data indicated that CD45RO+ Treg cells from pediatric asthma aberrantly increased intracellular Ca2+ concentrations following TCR activation compared with pediatric nonasthma controls. Conclusion: These impaired CD45RO+ Treg cell functions were correlated with asthma symptoms. The correlation was observed in the group with a highly expressed atopic phenotype and longer duration of asthma. We suggest that chronic inflammation in pediatric asthma airways may be the result of impaired regulatory functions of CD45RO+ Treg cells.

Background: Systemic lupus erythromatosus (SLE) is an autoimmune disease with 20–65% of patients developing lupus nephritis (LN). Studies have reported 10% of LN patients will end up with end stage renal disease and their mortality rate is higher compared to patients without LN. Abnormality of regulatory T cells (Tregs) level is thought to be a potential factor for this LN development. The aim of study was to evaluate the percentage of Tregs in LN patients.Methods: This was a comparative cross sectional study involving LN patients and age and gender matched controls with a 2:1 ratio. The patients were grouped into active and inactive LN based on their lupus activity index; complement levels, ANA, dsDNA antibodies, ESR, SLE Disease Activity Index (SLEDAI2K) score and also urine PCI (uPCI>0.05 for active group). Disease history, demographic data, routine blood test, peripheral blood for differentials count were taken and recorded. Peripheral blood mononuclear cells were stained with CD4, CD25 and Foxp3 antibodies and percentage of Tregs was analysed using BD fluorescence-activated cell sorting (FACS) cytometer. We compared demographic and laboratory parameters between healthy controls and LN patients as well as active and inactive LN patients.Results: A total of 34 LN patients (32 females, 2 males) were recruited. Their mean age and disease duration were 37.97±11.14 years and 110.95±65.07 months respectively. Thirteen matched controls with mean age 35.23±7.89 years were enrolled. There was no demographic difference between 2 groups of LN patients. Tregs were significantly lower in active LN compared to inactive LN and healthy control (0.44±0.37% vs. 1.89±0.46% vs. 3.12±0.56% of the CD4+, P

A lack of regulatory T (T(Reg)) cells that express CD4, CD25 and forkhead box P3 (FOXP3) results in severe autoimmunity in both mice and humans. Since the discovery of T(Reg) cells, there has been intense investigation aimed at determining how they protect an organism from autoimmunity and whether defects in their number or function contribute to the development of autoimmunity in model systems. The next phase of investigation - that is, to define the role that defects in T(Reg) cells have in human autoimmunity - is now underway. This Review summarizes our progress so far towards understanding the role of CD4(+)CD25(+)FOXP3(+) T(Reg) cells in human autoimmune diseases and the impact that this knowledge might have on the diagnosis and treatment of these diseases.

CD4+CD25+ Treg cells play a protective role in hypersensitivity pneumonitis by inhibiting IFN-γ-producing T cells. HP results from the repeated inhalation of environmental antigens; however, the roles of CD4+CD25+ Treg cells in HP are unknown. Therefore, we investigated the functions of CD4+CD25+ Treg cells in SR-induced murine HP. More severe HP was observed in CD4+CD25+ Treg cell-depleted mice than in control mice in terms of histological alterations, inflammatory cell numbers in BALF, and the serum level of SR-specific IgG, which were restored by the adoptive transfer of CD4+CD25+ Treg cells. The CD4+CD25+ Treg cell-depleted mice also showed elevated levels of IFN-γ, TGF-β, and reduced IL-4 production in the lungs. Moreover, IL-10 production of CD4+CD25+ Treg cells and direct contact between CD4+CD25+ Treg cells and CD4+ or CD8+ T cells in BALF resulted in reduced IFN-γ production. Taken together, CD4+CD25+ Treg cells play a protective role in SR-induced HP by suppressing IFN-γ production by T cells.

Background: Regulatory T-cell (Treg) modulation is one of the potential mechanisms of anti-tumour-necrosis-factor biological agents. However, literature data on psoriasis patients are lacking. Objective: To analyse the circulating CD4+CD25brightFOXP3+ subset in 30 patients with psoriasis vulgaris/arthropathic psoriasis treated with biologicals and to investigate its relationship with the clinical response. Methods: The CD25brightFOXP3+ expression within the CD4+ subset was determined by multi-parameter flow cytometry at baseline and during treatment. FOXP3 mRNA expression was analysed by real-time reverse transcription PCR. Results: A response was obtained in 16/17 patients (91.1%) with increased CD25brightFOXP3+ values and in only 3/11 patients (27.3%) who showed a CD25brightFOXP3+ decrease during biological treatment (p = 0.0001). Responders showed significantly higher values than did non-responders as from the first 2 months of treatment (p = 0.0032). A significantly higher posttreatment expression of mRNA FOXP3 was observed in responders compared to non-responders. Conclusion: Biological drugs induce a circulating Treg up-regulation in a significant percentage of patients; such an increase is an early predictive marker of response.

Background We showed previously that α-galactosylceramide (α-GalCer) treatment elevated splenic CD4 + CD25 + Foxp3 + T-cell numbers and suppressed the development of experimental allergic conjunctivitis (EC). Here, we investigated whether CD4 + CD25 + Foxp3 + T cells mediate the suppressive effects of α-GalCer treatment on EC. Methods To deplete CD4 + CD25 + Foxp3 + T cells, neonatal mice were thymectomized and intraperitoneally injected with anti-CD25 Ab. At 6 weeks of age, these mice were immunized with ragweed (RW) in aluminum hydroxide. Ten days later, the mice were challenged with RW in eye drops and 24 hours later, the conjunctivas and spleens were harvested for histological and flow cytometric analyses, respectively. α-GalCer or vehicle was injected 2 hours prior to RW challenge. In addition, α-GalCer was injected into thymus-intact EC-developing mice that had not been treated with anti-CD25 Ab. Results α-GalCer treatment significantly suppressed EC in the thymus-intact mice that had not been treated with anti-CD25 Ab. In contrast, α-GalCer treatment of thymectomized and anti-CD25 Ab-treated mice did not affect the severity of EC or splenic CD4 + CD25 + Foxp3 + T-cell numbers. However, α-GalCer treatment did significantly increase splenic CD4 + CD25 + Foxp3 + T-cell numbers in thymectomized mice that had not received anti-CD25 Ab. Conclusions α-GalCer treatment during the effector phase of EC increased CD4 + CD25 + Foxp3 + T-cell numbers, which in turn suppressed the development of EC.

Background Atopic dermatitis (AD) is a common chronic inflammatory skin disease of humans and dogs. Regulatory T cells (Tregs) are essential controllers of immune homeostasis and have been shown to play a key role in human AD, even though frequencies of Tregs in atopic human patients vary greatly. Only two studies have reported Treg numbers in the peripheral blood of dogs with canine AD (CAD). Objectives This study aimed to assess the numbers of circulating Tregs in healthy and atopic dogs, and to determine whether Treg numbers correlate with age, sex, disease severity or pre-treatment. Animals Client-owned dogs including 14 healthy dogs and 35 dogs with CAD. Methods Expression of Tregs in peripheral blood mononuclear cells was evaluated by flow cytometry. Tregs were phenotypically identified as T cells triple positive for CD4, CD25 and FoxP3. Results The percentage of circulating CD4+ CD25+ FoxP3+ Tregs in atopic dogs was increased significantly compared to healthy dogs (mean 2.1% versus 1%, P = 0.002) and correlated with disease severity (Pruritus Scale: r = 0.48, P = 0.003; CADESI-04: r = 0.34, P = 0.044). No significant differences in age or sex were found in either group and pre-treatment had no influence on results for atopic dogs. Conclusions Data suggest that, as in humans, CD4+ CD25+ FoxP3+ Tregs may contribute to the pathogenesis of CAD as indicated by an association between Treg frequency and disease severity. Further investigation is required to improve the understanding of the role of Tregs in atopic dogs. Resume Contexte La dermatite atopique (AD) est une maladie cutanee inflammatoire chronique frequente de l'homme et du chien. Les cellules T regulatrices (Tregs) sont essentielles dans le controle de l'homeostasie immunitaire et jouent un role cle dans l'AD de l'homme meme si la frequence de Tregs chez les patients atopiques humains varie de facon importante. Seules deux etudes ont etudiees le nombre de Tregs dans le sang peripherique du chien atopique (CAD). Objectifs Cette etude a pour objectif d'estimer le nombre de Tregs circulants chez le chien sains et atopique et de determiner si le nombre de Treg correle avec l'âge, le genre, la severite de la maladie ou le pretraitement. Sujets Des chiens de proprietaires dont 14 chiens sains et 35 chiens atopiques. Methodes L'expression des cellules mononuclees Tregs dans le sang peripherique a ete evaluee par cytometrie de flux. Les Tregs ont ete phenotypiquement identifies comme cellules T triplement positives pour CD4, CD25 et FoxP3. Resultats Le pourcentage de Tregs CD4+ CD25+ FoxP3+ circulants chez les chiens atopiques etait augmente significativement compare aux chiens sains (moyenne 2.1% versus 1%, P = 0.002) et correlait avec la severite de la maladie (echelle de prurit: r = 0.48, P = 0.003; CADESI-04: r = 0.34, P = 0.044). Aucune difference significative d'âge ou de genre n'a ete trouvee dans chaque groupe et le pretraitement n'avait aucune influence sur les resultats des chiens atopiques. Conclusions Les donnees suggerent que, comme pour l'AD humaine, les Tregs CD4+ CD25+ FoxP3+ peuvent contribuer a la pathogenie de la DAC telle que l'indique l'association entre la frequence de Treg et la severite de la maladie. De plus amples investigations sont necessaires pour ameliorer la comprehension du role des Tregs dans la dermatite atopique. Resumen Introduccion La dermatitis atopica (AD) es una enfermedad cronica inflamatoria de la piel comun en seres humanos y perros. Los linfocitos T reguladores (Tregs) son componentes esenciales del control de la homeostasis inmunitaria y han demostrar tener un papel clave de la dermatitis atopica humana, incluso aunque la frecuencia de los linfocitos T reguladores en los pacientes humanos con atopia varian enormemente. Solo dos estudios han publicado los numeros de linfocitos T reguladores en la sangre periferica de perros con dermatitis atopica canina (CAD). Objetivos este estudio esta enfocado a evaluar los numeros de linfocitos T reguladores en perros sanos y perros con atopia y determinar si los numeros de linfocitos T reguladores se correlacionan con la edad, sexo, severidad de la enfermedad o tratamiento previo. Animales perros de propietarios privados incluyendo 14 perros sanos y 35 perros con atopia. Metodos la expresion de linfocitos T reguladores en celulas mononucleares de sangre periferica se evaluo mediante citometria de flujo. Los linfocitos T reguladores fueron tipicamente identificados como celulas triples positivas para CD4, CD25, y FoxP3. Resultados el porcentaje de linfocitos T reguladores CD4+, CD25+ FoxP3+ en perros atopicos se incremento significativamente comparado con los perros sanos (media 2,1% frente a 1%, P = 0,002), y se correlaciono con la severidad de la enfermedad (escala de prurito: r = 0,48, P = 0,003; CADESI-04: r= 0,34, P = 0,044). No hubo diferencias significativas en edad basados en edad o sexo en ningun grupo y el tratamiento previo no tuvo influencia en los resultados en perros topicos. Conclusion e importancia clinica los datos sugieren que, al igual que en humanos, los linfocitos T reguladores CD4+, CD25+ FoxP3+ pueden contribuir a la patogenesis de la dermatitis atopica canina tal y como indica la asociacion entre la frecuencia de linfocitos T reguladores y la severidad de la enfermedad. Se requieren mas investigaciones para mejorar la comprension del papel de los linfocitos T reguladores en perros atopicos. Zusammenfassung Hintergrund Die atopische Dermatitis (AD) ist eine haufige chronische entzundliche Hauterkrankung bei Menschen und Hunden. Die regulatorischen T Zellen (Tregs) sind essentielle Kontrolleure der Homoostase des Immunsystems und es konnte gezeigt werden, dass sie bei der AD des Menschen eine wichtige Schlusselrolle spielen, obwohl die Anzahl der Tregs bei atopischen Menschen stark variieren. Es gibt bisher nur zwei Studien, die uber die Anzahl der Tregs im peripheren Blut von Hunden mit caniner AD (CAD) berichten. Ziele Das Ziel dieser Studie war eine Erhebung der Anzahl der zirkulierenden Tregs bei gesunden und atopischen Hunden, und festzustellen, ob die Anzahl der Tregs mit dem Alter, dem Geschlecht und der Schwere der Erkrankung oder mit einer Vorbehandlung korrelierten. Tiere Hunde im Privatbesitz, davon 14 gesunde Hunde und 35 Hunde mit CAD. Methoden Die Exprimierung der Tregs in peripheren mononuklearen Zellen des Blutes wurde mittels Flowzytometrie bestimmt. Die Tregs wurden phanotypisch als CD4, CD25 und FoxP3 positiv identifiziert. Ergebnisse Der Prozentsatz der zirkulierenden CD4+CD25+FoxP3+ Tregs bei atopischen Hunden war im Vergleich zu gesunden Hunden signifikant erhoht (Durchschnitt 2,1% versus 1%, P=0,002) und korrelierte mit der Schwere der Erkrankung (Juckreizskala: r=0,48, P=0,003; CADESI-04: r=0,34, P=0,044). Es bestanden in beiden Gruppen keine signifikanten Unterschiede zwischen dem Alter oder dem Geschlecht, eine Vorbehandlung hatte keinen Einfluss auf die Ergebnisse bei atopischen Hunden. Schlussfolgerungen Die Ergebnisse weisen darauf hin, dass wie beim Menschen CD4+CD25+FoxP3+ Tregs an der Pathogenese der CAD teilhaben, was sich durch den Zusammenhang der Anzahl der Tregs und der Schwere der Erkrankung zeigte. Eine weitere Untersuchung ist notig, um das Verstandnis der Rolle der Tregs bei atopischen Hunden zu erweitern.

Results TS patients exhibited a higher frequency of CD4FOXP3 Tregs among their lymphocytes (mean 2.06 vs. 1.52%, P = 0.005) and FOXP3 Tregs among their CD4 T cells (7.44 vs. 4.19%, P < 0.001) compared to controls. The expression of inhibitory CTLA-4 in the Tregs of TS patients was also significantly higher (mean fluorescence intensity = 214.1 vs. 184.6, P = 0.003). The frequency of Tregs expressing GITR, CXCR3, and CCR4CCR6 was comparable between the two groups. However, the ability of Tregs to suppress the in vitro proliferation of autologous CD4 CD25 T cells was significantly impaired in TS patients compared to controls (P < 0.05 at a 0.1:1 ratio of Tregs to target cells, P < 0.01 at 0.25:1, 0.5:1, and 1:1).

Autoantibodies induce various autoimmune diseases, including systemic lupus erythematosus (SLE). We previously described that CD4+CD25−LAG3+ regulatory T cells (LAG3+ Treg) are regulated by Egr2, a zinc-finger transcription factor required for the induction of T-cell anergy. We herein demonstrate that LAG3+ Treg produce high amounts of TGF-β3 in an Egr2- and Fas-dependent manner. LAG3+ Treg require TGF-β3 to suppress B-cell responses in a murine model of lupus. Moreover, TGF-β3- and LAG3+ Treg-mediated suppression requires PD-1 expression on B cells. We also show that TGF-β3-expressing human LAG3+ Treg suppress antibody production and that SLE patients exhibit decreased frequencies of LAG3+ Treg. These results clarify the mechanism of B-cell regulation and suggest therapeutic strategies., B cells reactive against self antigens can cause autoimmune disease, but are normally suppressed by regulatory T cells (Tregs). Here the authors show that a subset of Tregs can suppress lupus in a mouse model by making TGF-β3 cytokine and by engaging an inhibitory PD-1 receptor on B cells.

The aim of this prospective case control study is to determine CD4(+), CD25(+), and FoxP3(+) T regulatory cells (Tregs) and T helper cells (Ths) in obese, asthmatic, asthmatic obese, and healthy children. Obese (n = 40), asthmatic (n = 40), asthmatic obese (n = 40), and healthy children (n = 40) were included in this study. Blood samples collected from children were marked with CD4, CD25, ve Foxp3 in order to detect Tregs and Ths by flow cytometric method. Statistical analyses were performed. p ≤ 0.05 was chosen as meaningful threshold. Tregs exhibiting anti-inflammatory nature were significantly lower in obese (0.16 %; p ≤ 0.001), asthmatic (0.25 %; p ≤ 0.01), and asthmatic obese (0.29 %; p ≤ 0.05) groups than control group (0.38 %). Ths were counted higher in asthma group than control (p ≤ 0.01) and obese (p ≤ 0.001) groups. T cell immunity plays important roles in chronic inflammatory diseases such as obesity and asthma pathogeneses. Decreased numbers of Tregs found in obese, asthmatic, and asthmatic obese children might represent a challenge of these cells.