Temporal and spatial characterization of negative regulatory T cells in HIV-infected/AIDS patients raises new diagnostic markers and therapeutic strategies.

BACKGROUND: Negative regulatory T cells (Tregs) not only deplete effector T cells but also inhibit the clearance of HIV during infection, which may allow Tregs to be used as informative diagnostic markers. To facilitate both diagnosis and treatment, a thorough understanding of these regulators by characterizing them on temporal and spatial scales is strongly required. METHODS: Hundred HIV-infected/AIDS patients, including 87 males, with an average age of 35.8 years, as well as 20 healthy controls, were enrolled. Flow cytometry was used to analyze CD3+T cells, CD4+T cells, and CD8+T cells to evaluate the immune status of the participants. Then, a group of representative negative regulatory T cells, including CD4+PD-1+T cells, CD4+PD-1high T cells, CD8+PD-1+T cells, and CD4+CD25high Tregs was also analyzed to explore their effects on disease progression and intercorrelation. RESULTS: The percentages of CD4+ PD-1+ T cells and CD4+ CD25high Tregs increased in patients with the same ultrahigh significance. Temporally, the patients with both intermediate-stage and late-stage disease had higher percentages of CD4+ PD-1+ T cells; however, the percentage of CD4+ CD25high Tregs only increased in the patients with late-stage disease. In addition, CD4+ PD-1+ T cells but not CD4+ CD25high Tregs were negatively correlated with the absolute CD4+ T cell count. Spatially, no correlations between CD4+ PD-1+ T cells and CD4+ CD25high Tregs were observed, which suggests these Tregs function differently during immunosuppression. CONCLUSIONS: This study characterized negative regulatory T cells in HIV-infected/AIDS patients at both temporal and spatial scales and found that CD4+ CD25+ Tregs and CD4+ PD-1+ T cells could be used as potential diagnostic markers for identifying different disease stages and monitoring disease progression.

The role of soluble programmed death protein-1 (sPD-1) and soluble programmed death ligand-1 (sPD-L1) in rat corneal transplantation rejection.

BACKGROUND: Immunological rejection is one of the problems in corneal transplantation. Recently, some research found out that soluble programmed death protein-1 (sPD-1) and soluble programmed death ligand protein-1 (sPD-L1) play a significant role in immunologic suppression. OBJECTIVES: To explore expression of sPD-1 and sPD-L1 in a penetrative corneal transplantation model and its relationship with transplant rejection. MATERIAL AND METHODS: Autologous corneal transplantation rat models and allogeneic corneal transplantation rat models were used as the control group and the experimental group, respectively. Changes of the transplanted grafts were observed under a slit-lamp microscope. Hematoxylin-eosin (H&E) staining was applied to examine the histopathological features of the corneal grafts. Flow cytometry was used to analyze CD4+CD25+Treg in the serum and spleen. The sPD-1, sPD-L1, interleukin 10 (IL-10) and interleukin 4 (IL-4) levels in serum and the aqueous humor of the rats were detected using enzyme-linked immunosorbent assay (ELISA). RESULTS: After the operation, no transplant rejection occurred in the control group. Flow cytometry results showed that expressions of CD4+CD25+Treg in serum in the experimental group were lower than those in the control group (p < 0.05). The ELISA results showed that after the operation, sPD-1 and sPD-L1 expression levels in serum in the experimental group were higher than in the control group (all p < 0.05). After the operation, lL-10 and IL-4 content in serum in the experimental group was lower than in the control group (all p < 0.05). The sPD-1/sPD-L1 ratio in the experimental group was higher than in the control group. CONCLUSIONS: Increases of sPD-1 content and decreases of CD4+CD25+Treg, IL-10 and IL-4 levels may be involved in corneal allograft rejection. Dynamic detection of the content of sPD-1 and sPD-L1 in serum and aqueous humor after the operation would help in understanding the local immune response in a clinical setting and predicting the occurrence of corneal graft rejection.

Exposure to silver nanoparticles induces immunological dysfunction in pregnant mice.

The adverse outcomes of silver nanoparticles (AgNPs) on pregnancy have been studied in murine animals. However, the potential toxicity of AgNPs to immune balance, which is essential for maintaining a normal pregnancy, still requires further exploration. Therefore, this study assessed the effect of AgNPs on the immune balance during gestation time. Pregnant mice were given a dose of 1 mg/kg of AgNPs and silver ion on gestation days 3.5 to 9.5 by tail vein injection. Results showed that the AgNPs and silver ion decreased the number of CD4+ CD25+ Treg cells which were the important cells in the immune system, thereby disrupting the balance of normal immune tolerance function, activated the inflammatory responses, together with the reductive production of placental immunoregulatory genes, and the expression of inflammatory factors in the placenta in the Ag-treated groups increased. These effects increased the absorption rate. Furthermore, the inflammatory signaling pathway p38MAPK/AP-1/MMP-9 in the placenta was activated, indicating that Ag induced inflammation through this signaling pathway. All results indicated that undesirable pregnancy outcome caused by AgNPs could be happened by stimulating immunological dysfunction. Therefore, the potential risks to embryogenesis exposure to AgNPs that caused immune imbalance should be given sufficient attention.

Viral IL-10 promotes cell proliferation and cell cycle progression via JAK2/STAT3 signaling pathway in nasopharyngeal carcinoma cells.

Epstein-Barr virus (EBV) is positively related to the morbidity of nasopharyngeal carcinoma (NPC) in Asia. After infection, EBV can produce several proteins, including viral interleukin-10 (vIL-10). But the mechanism by which vIL-10 contributes to NPC cell proliferation and cell cycle progression is not well understood. In this study, EBV negative and positive cell lines, and the JAK2/STAT3 signal pathway inhibitor AG490 were used to illustrate the role of vIL-10 in NPC. Cell proliferation and cell cycle were measured by CCK-8 and flow cytometry. The expression levels of related protein were measured by Western blotting. High concentrations of vIL-10 and IL-6 were found in the EBV positive patients. The expression level of IL-6 was positively related to the presence of concentration of vIL-10. vIL-10 can promote cancer cell proliferation and G1 to S phase transmission via upregulating the IL-6 protein level by activating the JAK2/STAT3 signal pathway. Furthermore, EBV can induce the formation of cytotoxic T cells, whereas vIL-10 can block the function of cytotoxic T cells. Taken together, these results suggest that vIL-10 promotes cell proliferation and cell cycle progression via JAK2/STAT3 signaling pathway in NPC.

Eplerenone Reverses Cardiac Fibrosis via the Suppression of Tregs by Inhibition of Kv1.3 Channel.

Background: Fibroblast proliferation is a critical feature during heart failure development. Previous studies reported regulatory T-lymphocytes (Tregs)' protective role against myocardial fibrosis. However, notably, Tregs also secrete fibrogenic cytokine TGF-ß when activated. This study aimed to clarify the intriguing link between Tregs and fibrosis, the role of Tregs Kv1.3 potassium channel (regulating T-lymphocytes activation) in the fibrosis process, and how selective aldosterone receptor antagonist Eplerenone affects Tregs and fibrosis through its action on Kv1.3 channel. Methods and Results: After co-incubation with Tregs, cardiac fibroblast proliferation (CCK-8 assay) and levels of collagen I, III, and Matrix metalloproteinase2 (ELISA) significantly elevated. Cell viability assays, Kv1.3 channel mRNA (RT-qPCR), and protein expression (In-Cell Western Blotting) revealed Tregs were activated/proliferated when co-cultured with fibroblasts. Treg intracellular TGF-ß level increased by 5.8-fold, far more than that of intracellular IL-10, extracellular TGF-ß and IL-10 (ELISA). And 30 µM eplerenone suppressed Tregs proliferation by 82.77% and furthermore, suppressed intracellular TGF-ß level to a significantly greater extent than that of intracellular IL-10, extracellular TGF-ß and IL-10. Moreover, the Kv1.3 current (whole-cell patch clamp) of Tregs in congestive heart failure patients and rats (induced by coronary artery ligation and exhaustive exercise) elevated by >4-fold than that of healthy volunteers and control rats, whereas 30 µM eplerenone suppressed the current by >60% in control Tregs. In addition, docking calculations (AutoDock software 4.0 suite) showed eplerenone has higher H-bond energy with Kv1.3 channel than other selective blockers. Conclusion: Immuno-regulation in the late stage of CHF activates Tregs proliferation via the upregulation of Kv1.3 channels, which promotes cardiac fibrosis by primarily secreting TGF-ß. Taken together, eplerenone's high affinity to Kv1.3 channel enables it to antagonize the Kv1.3 channels directly to suppress Tregs proliferation, which in turn may play an immuno-regulatory role during CHF.

Depletion of FoxP3+ Tregs improves control of larval Echinococcus multilocularis infection by promoting co-stimulation and Th1/17 immunity.

INTRODUCTION: The growth potential of the tumor-like Echinococcus multilocularis metacestode (causing alveolar echinococcosis, AE) is directly linked to the nature/function of the periparasitic host immune-mediated processes. Previous studies had shown that regulatory T cells (Tregs) become gradually up-regulated in the course of both chronic human and murine AE. Thus we now tackled the role of FoxP3+ Tregs and FoxP3+ -Treg-regulated immune response in contributing to the control of this helminthic infection. METHODS: The infection outcome in E. multilocularis-infected DEREG mice was measured upon determining parasite load (wet weight of parasitic metacestode tissue). Flow cytometry and qRT-PCR were used to assess Treg, Th17-, Th1-, Th2-type immune responses and antigen presenting cell activation. RESULTS: We showed that E. multilocularis-infected DEREG-mice treated with DT (as compared to infected control DEREG-mice without DT application) exhibited a significantly lower parasite load, associated with a persisting capacity of co-stimulation, and an increased Th1/Th17-polarization. CONCLUSIONS: FoxP3+ Tregs appear as one of the key players in immune regulatory processes favoring (i) metacestode survival by inhibiting the maturation potential of co-stimulatory activity and (ii) T cell exhaustion (suppressing Th1/Th17-type immune responses). We showed as well that prospectively, targeting FoxP3+ Tregs could be an option to develop an immunotherapy against AE.

Isolation of Immune Cells for Adoptive Transfer.

Adoptive transfer of T lymphocytes is a useful technique to characterize the role of the immune system in hypertension and vascular disease. Here we describe as an example the isolation of splenic T regulatory cells from donor mice processed to obtain a single cell suspension, followed by negative and positive selection to obtain CD4+ T cells and CD4+CD25+ Treg cells, respectively. Treg cells can be subsequently transferred to recipient animals.

Evaluation of PLGA containing anti-CTLA4 inhibited endometriosis progression by regulating CD4+CD25+Treg cells in peritoneal fluid of mouse endometriosis model.

Our study investigated poly(lactic-co-glycolic acid) (PLGA) as protein delivery vehicles encapsulate CTLA-4-antibody (anti-CTLA-4) which is essential for CD4+CD25+Treg cells suppressive function exposing superior potential for inhibiting endometriosis progress in mouse model than single anti-CTLA-4. Anti-CTLA-4 loaded PLGA combined to ligands CTLA-4 in surface of CD4+CD25+Treg cells which distributed in peritoneal fluid of mouse endometriosis model. The particle size, zeta potential of the anti-CTLA-4 loaded nanoparticles was detected by dynamic light scattering. Morphology of nanoparticles was evaluated by transmission electron microscopy (TEM). Confocal laser scanning microscopy (CLSM) indicated distribution of anti-CTLA-4 with PLGA or without in peritoneal fluid. Cumulative anti-CTLA-4 release from nanoparticles was evaluated by Micro BCA assay. The percentage of CD4+CD25+Treg cells in peritoneal fluid was demonstrated by flow cytometer. In vitro experiment we co-culture ectopic endometrial cells (EEC) with isolated CD4+CD25+Treg cells in peritoneal fluid (PF), proliferation and invasion of ectopic endometrial cells (EEC) was measured by BrdU ELISA assay and Matrigel invasion assay. In comparison with anti-CTLA-4 without nanoparticles, the bioconjugates PLGA/anti-CTLA-4 were tolerated in peritoneal fluid with a controlled release of anti-CTLA-4 in 3, 7, 14days. Moreover, PLGA/anti-CTLA-4 had superior protective regulation ability to reduce level of CD4+CD25+Treg cells in peritoneal fluid. Most strikingly, in vitro experiment, PLGA/anti-CTLA-4 exhibited better ability in inhibiting proliferation and invasion of ectopic endometrial cells in co-culture system compared with anti-CTLA-4. Progressively, PLGA/anti-CTLA-4 had better suppressive activity to inhibited IL-10 and TGF-beta secreted by CD4+CD25+Treg cells which indicating that PLGA/anti-CTLA-4 suppressed cells proliferation and invasion through reduced IL-10 and TGF-beta production. Thus, PLGA/anti-CTLA-4 may be a potential strategy for endometriosis therapy.

Experimental study on the expression of T cells and cytokines in the regulation of autoimmune dry eye / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the role of regulatory T cells in the pathogenesis of autoimmune dry eye and to analyze the expression of endogenous regulatory T cells and related cytokines in the lacrimal system to provide a more effective treatment for autoimmune dry eye disease treatment programs. <p>METHODS: Rabbit lacrimal gland epithelial cells isolated and cultured for a period of time were separately cultured and mixed with isolated peripheral blood lymphocytes in a ratio of 1: 1 and cultured with 5-Bromo-2-deoxyUridine(BrdU)to detect the proliferation of peripheral blood lymphocytes, and the activated autologous peripheral blood lymphocytes were transferred to donor rabbits via ear vein. The rabbits were used as the rabbit model of autoimmune dry eye and the normal rabbits which did not induce the disease as the control group. Before and after the experiment, tear film rupture time and tear secretion at 2,4,6,8wk,were observed. At 4 wk after the transfusion the rabbits were sacrificed for the collection of bilateral upper and lower lacrimal gland and conjunctiva, and pathological HE staining, and then extract the total RNA in lacrimal gland. The expression of IL-17, TNF-α, IL-6 and TGF-γ was detected. <p>RESULTS: BrdU detected peripheral lymphocyte proliferation rate of 3.72. Compared with the normal group, the time of rupture of the tear film decreased(<i>P</i><0.05), and the tear of the model group was significantly lower than that of the control group(<i>P</i><0.05). The expression of IL-17, TNF-α, TGF-γ and IL-6 in the model group were significantly higher than those in the control group(<i>P</i><0.05). The expression of IL-10 and TGF-β was significantly lower than those of the control group(<i>P</i><0.05). The ratio of CD4⁺ CD2⁺ Treg cells in lacrimal gland tissue and spleen was higher in control group. The proportion of CD4⁺CD25⁺ Treg cells in model group decreased(<i>P</i><0.05). <p>CONCLUSION: IL-17, TNF-α, IL-6 and TGF-γ play important roles in inflammatory reaction of lacrimal gland and the immunodepression active by CD4⁺CD25⁺Treg cells decreases when the rabbit model of dry eye is involved.

Effects of vasoactive intestinal peptide on the ratio of CD4+CD25+Treg/CD4+T cell and the expression of TGF-β1 in EAE rats / 西安交通大学学报(医学版)

Objective To explore the effects of vasoactive intestinal peptide (VIP)on the ratio of CD4+CD25+Treg/CD4+T cell and the expression of TGF-β1 in experimental autoimmune encephalomyelitis (EAE)rats. Methods We randomly divided 60 healthy female Wistar rats into normal control group,EAE control group,VIP low-dose group and VIP high-dose group.We used myelin basic protein (MBP)+ complete adjuvant (CFA)to establish EAE model. Since the day of model construction, the low- and high-dose VIP groups received intraperitoneal injection of 4 nmol/kg (0.2 mL)and 16 nmol/kg (0.8 mL)of VIP every other day,respectively;normal control group and EAE group received injection of saline of 0.8 mL for 10 days in a row.We recorded the peak of neurological dysfunction score (NDS)changes in the rats,observed the pathological changes and GFAP+astrocyte activation in the brain at the morbidity peak of rats with HE staining,and detected the ratio of CD4+CD25+Treg/CD4+T in the spleen with FACS and TGF-β1 cytokine level in brain tissue with ELISA.Results The peak nerve dysfunction score was decreased in each VIP dose group.In normal control group,there were decreased inflammatory cell infiltration and decreased number of active astrocytes in the brain tissue.The degree of infiltration of inflammatory cells and astrocyte activation in VIP control groups were significantly lower than those in EAE group.The CD4+CD25+Treg/CD4+T cell ratio of the spleen tissue in each dose VIP treated group rats was higher than that in EAE control group.The cytokine level of TGF-β1 in the brain tissue increased in each VIP dose group in the dose-dependent manner.Conclusion Through up-regulating the ratio of CD4+CD25+Treg/CD4+T cell in the spleen tissue,increasing TGF-β1 content in brain tissue,and inhibiting the infiltration of inflammatory cells and the astrocyte activation,VIP plays an important role in prevention and control of EAE.

Cimetidine down-regulates stability of Foxp3 protein via Stub1 in Treg cells.

Foxp3-expressing Treg cells have been well documented to provide immune regulation by promoting immune tolerance and suppressing immune over-reaction. Cimetidine (CIM), used to inhibit stomach acid secretion, has been reported to promote immune responses and suppress Treg cell function in several studies. However, the underlying mechanism is unknown. To investigate CIM effects on the suppressive function of Treg and Foxp3, here we used CIM to stimulate human CD4+CD25+ Treg cells and Jurkat T cells and evaluated changes of Foxp3 expression and stability. Our data showed that CIM leads to a reduction of Foxp3 via E3 ligase Stub1-mediated proteosomal degradation, which is dependent on an activated PI3K-AKT-mTOR pathway. Thus, CIM affects the suppressive function of Treg cells by destabilizing their Foxp3 expression.

Short-Chain Fatty Acids Regulate Cytokines and Th17/Treg Cells in Human Peripheral Blood Mononuclear Cells in vitro.

Short-chain fatty acids (SCFAs) have been recognized as mediators of immune responses, including pathways of cytokine production. In this study, we investigated the immune-regulatory effects of SCFAs on human peripheral blood mononuclear cells (PBMCs) from buffy coat of healthy donors. PBMCs were exposed to varying concentrations of individual SCFAs or of their mixtures of acetate, propionate and butyrate. The productions of interleukin (IL) IL-1ß, IL-2, IL-6, IL-10, IL-17, IL-21, IL-23 and transforming growth factor beta 1 (TGF-ß1) were assessed. T cell differentiation after exposure to SCFAs was also examined. Compared with lipopolysaccharide (LPS)-stimulated cells (controls), SCFAs slightly decreased TGF-ß1 production and reduced IL-6 production; butyrate was more effective than acetate or propionate. SCFAs particularly butyrate caused the induction of CD4+CD25+ regulatory T cells (Treg) rather than Th17 cells. SCFAs may up-regulate the production of anti-inflammatory cytokines in PBMCs, resulting in the induction of CD4+CD25+ Treg cells.

The expression and significance of CD4+CD25+ regulatory T cells in the peripheral blood of patients with systemic sclerosis / 中华风湿病学杂志

Objective To study the phenotype and function of CD4 +CD25 + Treg cells in the peripheral blood of patients with systemic sclerosis (SSc) and their relationship with fibrosis.Methods The proportion of Foxp3, CD127, CTLA-4 and CD69 on CD4+CD25+ Treg cells in peripheral blood were detect by flow cytometry;the levels of TGF-[1 and IL-10 in serum were detect by enzyme-linked immunosorbent assay (ELISA) in patients with SSc.The correlation between Treg cells and the score of chest HRCT, MRSS, and disease activity was analyzed.T test and Pearson correlation analysis were used for statistical analysis.Results ① Compare to the control group, the proportion of CD4+CD25+ Treg cells in peripheral blood of SSc patients was increased significantly(12.9±2.4 vs 14.9±2.2, t=2.63, P=-0.012), and the expression of CD69+, CTLA-4+ on CD4+CD25+ Treg cells was decreased significantly (P＜0.01).② Compare to the control group, the proportion of CD4+CD25+Foxp3 + cells and CD4+CD25+CDI27-cells in peripheral blood of SSc patients was increased significantly (respectively, 3.3±0.7 vs 5.0±0.7, 5.1±1.6 vs 7.6±2.0, t=7.03, 4.195;P＜0.01), but no correlation between them was detected.③ The level of TGF-β1 in the serum of the SSc patients was lower than that of the control group(86±29 vs 133±29 ng/ml, t=-5.026, P=0.000).However, IL-10 had no significant difference between the two groups.④ The proportion of CD4+CD25 +Foxp3 + cells and CD4~D25 +CD127-cells in peripheral blood of SSc patients was positively correlated with the scores of chest HRCT (respectively, r=-0.541, P=0.02;r=0.486, P=0.041), and no correlation was observed with ESR, CRP.In addition, CD4+CD25+Foxp3+ cells were associated with MRSS.Conclusion The proportion of CD4+CD25+ Treg cells in the peripheral blood of SSc patients is increased, but they alters the immune function.The different phenotypes of Treg cells of CD4+CD25+Foxp3+ cells and CD4+CD25+CD127-cells in peripheral blood of SSc patients are increased significantly, which changes along with skin and lung fibrosis.The associated cytokine TGF-β1 is reduced, and IL-10 is not significantly changed.

Characteristics of the peripheral T cell immune response of patients at different stages of vascular cognitive impairment.

AIM: To investigate the characteristics of the peripheral T cell immune response of patients at different stages of vascular cognitive impairment (VCI). METHODS: 61 Arterial atherosclerotic cerebral infarct induced VCI patients, including 28 vascular dementia (VaD) cases, 33 no dementia (VCI-ND) cases, and 25 atherosclerotic cerebral infarct patients with normal cognitive function (CI-NC) as controls were enrolled. Peripheral CD8(+)T, CD4(+)CD25(+) Treg, CD4(+)IL-17(+) Th17 cells proportion, and IL-1ß, IL-2, IL-6, IFN-γ levels, and neuropsychological function were assessed. RESULTS: There was no difference in average age, gender ratio, years of education, and risk factors of infarct among the three groups. Peripheral CD4(+)CD25(+) Treg in VCI-ND and VaD groups were significantly lower than that in controls, and CD8(+) T cells were markedly elevated in VaD group. The IL-17(+) Th17 cell proportion did not differ significantly among three groups. IL-6 and IFN-γ expression levels in VaD group were higher than those in other two groups. The VDAS-Cog executive function subscale score was negatively correlated with CD4(+)CD25(+) Treg proportion in VCI patients, and positively correlated with IL-6 levels. CONCLUSION: VCI patients demonstrated a decrease in peripheral CD4(+) Treg proportion and increased IL-6 expression, and both parameters were correlated with the decline of executive functions.

Simvastatin combined with aspirin increases the survival time of heart allograft by activating CD4(+)CD25(+) Treg cells and enhancing vascular endothelial cell protection.

OBJECTIVE: The objective was to investigate whether the combination of simvastatin and aspirin treatment prolongs the survival time of the heart allograft in rat and its underlying mechanism. METHODS: Heterotopic heart transplantation was performed using Wistar rats as donors and Sprague-Dawley (SD) rats as recipients. The SD rats were randomly divided into five groups (n=20/group): sham, HT (heart transplantation), HT+simvastatin (HT+S), HT+aspirin (HT+A), and HT+aspirin+simvastatin (HT+A+S). After transplantation, at 3, 7, 10, 15, 20, 30, and 40 days, the endothelial nitric oxide synthase (eNOS) expression was assessed by immunohistological staining; nitric oxide (NO) levels were analyzed by Griess assay; the activation of CD4(+)CD25(+) T regulatory lymphocytes (Tregs) was analyzed by flow cytometry; and pathological changes in the graft heart were determined by histology. RESULTS: Combined treatment of hearts with simvastatin and aspirin significantly prolonged the mean survival time of heart allografts [8 ± 1.2 days (n=18), 20 ± 3.4 days (n=19), 21 ± 2.8 days (n=19), and 39 ± 5.3 days (n=19) for HT, HT+S, HT+A, and HT+A+S group, respectively; HT vs. HT+A+S, P<.001; HT vs. HT+S or HT+A, P<.05]. In addition, CONCLUSIONS: Simvastatin given in combination with aspirin delayed the development of pathological changes in the myocardium, reduced vascular damage and prolonged the survival time of cardiac allograft. The underlying mechanism is linked with CD4(+)CD25(+)-Treg-cell-induced immune tolerance and enhanced vascular endothelial cell protection.

Expression of Th17, CD4 +CD25 +Treg, HLA-DR mRNA in peripheral blood of children with hand, foot and mouth diseases / 中华临床感染病杂志

Objective To investigate the expressions of Th17, CD4 +CD25 +Treg, HLA-DR mRNA in peripheral blood of children with EV71-induced hand, foot and mouth diseases ( HFMD ) and their clinical significance.Methods Stratified random sampling was used to select 60 children with HFMDs from Liaocheng People’s Hospital from February to October, 2014, including 20 mild, 20 severe and 20 critically ill cases.Twenty healthy children were also enrolled as the control group.All the children with HFMDs were given ribavirin (10 mg/kg) for the treatment.The percentages of Th17 and CD4 +CD25 +Treg cells in CD4 +T cells of peripheral blood were determined by flow cytometry, and the expression of HLA-DR mRNA in peripheral blood mononuclear cells was detected by reverse transcription polymerase chain reaction ( RT-PCR).Analysis of variance and SNK-q test were used to compare the expressions of Th17, CD4 +CD25 +Treg and HLA-DR mRNA among groups, and Pearson correlation analysis was performed to reveal the correlations between HLA-DR mRNA and Th17, CD4 +CD25 +Treg.Results Compared with the control group, the expression of Th17 was increased, while CD4 +CD25+Treg and HLA-DR mRNA expressions were decreased in children with HFMDs on d1 of treatment (F=310.4, 81.5 and 545.4, P0.05), but Th17 level in fatal was still higher and CD4 +CD25 +Treg level was still lower than those in control group (t=16.4 and 12.0, P<0.05).After 10 d of treatment, HLA-DR mRNA expressions in mild group and severe group were increased to the normal level.HLA-DR mRNA expression in surviving patients of critical ill group was still lower than that in mild group and severe group (P<0.05), but was higher than that in fatal patients (t=7.8, P<0.05).Pearson correlation analysis showed that, HLA-DR mRNA was negatively correlated with Th17 level (r=-0.770, P<0.01), and positively correlated with CD4 +CD25 +Treg level (r=0.883, P<0.01).Conclusion The expressions of Th17, CD4 +CD25 +Treg cells, and HLA-DR mRNA are correlated with the severity of HFMD, and may be used for evaluation of disease severity and prediction of disease outcomes.

Effect of simvastatin combined with aspirin on heart allograft / 实用医学杂志

Objective To observe the effects of simvastatin combined with aspirin on the heart allograft and detect its mechanism. Methods Heterotopic heart transplantation was performed from Wistar to Sprague-Dawley (SD) rats. All SD rats were randomly assigned to Sham; HT (heart transplantation); HT + simvastatin(HT + S);HT + aspirin (HT + A); HT + aspirin + simvastatin (HT + A + S) group at different time points (day 3, 7, 10, 15, 20, 30 and 40) after transplantation (n=20). eNOS expression was detected by immunohistochemical methods and NO levels was measured by Griess assay. Meanwhile , the analysis of CD4+CD25+Tregs was performed by flow cytometry and histological examination for pathological change of heart and vascular. Results Simvastatin combined with aspirin significantly prolonged the mean survival time of heart allografts in HT + A + S group to (39 ± 5.3) days (n = 19) and in HT group to (8 ± 1.2) days (n = 18) (P < 0.001); simvastatin combined with aspirin delayed pathological changes of the transplanted hearts and protected vascular damage; simvastatin combined with aspirin upregulated eNOS and enhanced NO secretion. The level of CD4+CD25+Tregs in the blood of HT + A + S rats was significantly increased (2.2 ± 0.5)%, (2.9 ± 0.8)%, (4.3 ± 1.0)%, (8.3 ± 1.7)% and (14.3 ± 3.7)% for sham, HT, HT + S, HT + A and HT + A + S group respectively, HT vs. HT + A (P < 0.05) or HT + A + S (P < 0.01). Conclusion Simvastatin combined with aspirin delays the development pathology of myocardial and vascular damage and prolongs the survival time of cardiac allograft. The responsible mechanism is associated with activating CD4+ CD25+ Treg cells induction immune tolerance and enhancing vascular endothelial cell protection.

Influence of coix seed triglyceride combined with transcatheter arterial chemoembolization therapy on the immune function in patients with advanced primary hepatocellular carcinoma / 介入放射学杂志

Objective To investigate the influence of coix seed triglyceride combined with transcatheter arterial chemoembolization (TACE) therapy on AFP, CD4﹢CD25﹢regulatory T (Treg) cells and cellular immune function in patients with advanced primary hepatocellular carcinoma (HCC). Methods A total of 50 patients with inoperable HCC, whose imaging examination showed no distant metastasis, were divided into the study group (n=25) and the control group (n=25). Coix seed triglyceride together with TACE was employed for the patients of the study group, while only TACE was adopted for the patients of the control group. For the patients of the study group, transcatheter hepatic artery infusion of 100 ml coix seed triglyceride was carried out during the performance of TACE, and postoperative intravenous drip of coix seed triglyceride (200 ml/d) was used for 5 days. The peripheral blood samples were collected one week before and one month after the treatment to detect the changes of AFP and T lymphocyte subsets (CD3﹢, CD4﹢, CD8﹢,CD4﹢/CD8﹢ and Treg) levels. One month after the treatment, enhanced CT, MRI or PET-CT was performed to evaluate the necrosis degree of the tumor. Results After the treatment, AFP levels was decreased in both groups, when compared the preoperative data the differences were statistically significant (P0.05). In the study group, the percentage of Treg cells decreased from preoperative (8.27±6.65)%to postoperative (4.22± 1.59)%, the difference was statistically significant (P0.05). Conclusion In treating advanced primary HCC, coix seed triglyceride combined with TACE can reduce the percentage of Treg cells, thus, influence the patient’s cellular immune status and possibly decrease the recurrence rate of HCC after TACE therapy.

Effects of leukotriene receptor antagonists on airway remodeling and Th17 cells/CD4+CD25+regulatory T cells expresson in asthma / 临床儿科杂志

The airway remodeling of bronchial asthma is the result of repeated airway inlfammation. Its occurrence is a complex process involving many cytokines, inlfammatory mediators and associated cellular components, of which leukotrienes are important mediators of inlfammation in the airway remodeling. Many factors inlfuence Airway remodeling. In recent years, effects of Th17 cells and CD4+CD25+regulatory T cells (CD4+CD25+treg cells) on airway remodeling is growing in importance. Leukotriene receptor antagonist is an effective drug in the treatment of asthma and can suppress airway remodeling. But the exact mechanisms and its impact on the proportion of Th17 cells/CD4+CD25+treg cells is not yet clear. Therefore, the clariifcation of the changes of Th17 cells/CD4+CD25+treg cells expression in airway remodeling and the speciifc pathways, biological effects, inlfuence of the proportion of Th17 cells/CD4+CD25+treg cells expression after leukotriene receptor antagonist intervene can provide a new target for prevention and the treatment of asthma in the future.

Recent Research in Role of CD4+CD25+Regulatory T Cell in Pathogenesis of Immune Thrombocytopenia / 天津医药

Immune thrombocytopenia (ITP), the most common hemorrhagic disease, is an organ-specific autoimmune disease characterized by decreased platelets count due to auto-antibodies mediating platelets destruction and insufficient platelets production. The etiology of ITP is still not completely known. Regulatory T cells, also called Tregs, are character-ized by CD4+CD25+, and positive of transcription factor forkhead box P3. They belong to a subpopulation of T cells special-ized for immune regulation and play an important role in maintaining the immune balance. Decreased production and defect-ed function of CD4+CD25+Treg was found not only in the ITP animal model but also in the ITP patients. It indicates that the Treg was involved in the pathology of ITP. This review focus on the characteristic and function of Tregs and their relationship with pathogenesis of ITP.