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Abstract Introduction/objectives Systemic lupus erythematosus (SLE) is a classic prototype of the multisystem autoimmune disease and follows a relapsing and remitting course. Triptolide is a diterpene triepoxide extracted from Chinese medicine Tripterygium wilfordii Hook F, with potent immunosuppressive and anti-inflammatory properties. Our previous work observed that triptolide alleviated lupus in MRL/lpr lupus mice with the upregulation of regulatory T cells (Treg) proportion in previous study. In this study, we explored the proportion of follicular T regulatory (Tfr), follicular T helper (Tfh) and germinal center (GC) B cells in lupus mice and evaluated the efficacy of triptolide for lupus treatment in vivo. Methods 20 female MRL/lpr mice were randomly divided into 2 treatment groups and treated orally with vehicle or triptolide. C3H mice were all housed as controlled group and treated orally with vehicle. The percentage of Tfr cells, Tfh cells and GC B cells in spleen of mice were detected by Flow cytometric analysis and immunohistochemistry after 13 weeks of treatment. Results We found that the percentage of Tfr cells decreased in MRL/lpr mice compared with controlled mice. The percentage of Tfh cells in MRL/lpr mice was significantly higher compared with that in controlled mice. The ratio of Tfr/Tfh is also decreased in lupus mice. After treated with triptolide in MRL/Lpr mice in vivo, the percentage of Tfr cells and ratio of Tfr/Tfh increased. The proportion of GC B cells also decreased in mice treated with triptolide by FACS and immunohistochemistry. Conclusions Our results demonstrate that the effect of triptolide in alleviating lupus is partly by reversing immune imbalance with increased percentage of Tfr cells and ratio of Tfr/Tfh. Triptolide might also has effect on immune response through inhibiting proliferating GC B cells.
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Objective:The present study was to observe the effect of Qianjin Weijingtang on the differentiation of helper T cells 17 （Th17）/T regulatory cell （Treg） and the expressions of related cytokines in the lung tissues of the model rats exposed to cigarette smoke. Method:Totolly 60 male rats were randomly assigned into six groups （control group， model group， acetylcysteine group and Qianjin Weijingtang high， moddle and low dose groups）， with 10 rats in each group. After 30 day's modeling and 30 day's intervention， rats were killed peacefully with their tissues collected. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression of retinoic acid associated orphan receptor （ROR-γt） and forkhead/pterygoid helix transcription factor 3 （Foxp3） mRNA， enzyme-linked immunosorbent assay （ELISA） was used to check the concentration of interleukin-17（IL-17）， IL-6， IL-10 and transforming growth factor-β1（TGF-β1）. Htoxylin eosin （HE） staining was used to observe the pathological changes of lung tissues， while flow cytometry was used to detect Treg（Foxp3+CD25+）， Th17（CD4+IL-17+） and Treg/Th17 ratio. Result:As compared with the control group， the ROR-γt mRNA expression in model group was higher（P<0.01）. As compared with the model group， various doses of Qianjin Weijingtang down-regulated the expression（P<0.05，P<0.01）. As compared with the control group， the expression of the Foxp3 mRNA was down-regulated in model group（P<0.05）， but was up-regulated in Qianjin Weijingtang middle and low dose groups（P<0.05）. Compared with the control group， the concentration of IL-17 and IL-6 in the model group were significantly increased， while the concentration of IL-10 and TGF-β1 in the model group were significantly decreased（P<0.05，P<0.01）. As compared with the model group， intervention with various doses of Qianjin Weijingtang could help to decrease the concentration of IL-17 and IL-6 in lung tissues， and increase the concentration of IL-10（P<0.05）， which were consistent with those of Real-time PCR results. Flow cytometry examination showed that the Th17（CD4+IL-17+） proportion decreased and Treg/Th17 ratio increased after Qianjin Weijing Tang middle and low dose intervention（P<0.05，P<0.01）. Conclusion:Qianjin Weijingtang could regulate Treg/Th17 ratio and help to achieve Th17/Treg balance.
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The ability of nematodes to manipulate the immune system of their host towards a Th2 and T regulatory responses has been proposed to suppress the inflammatory response. Clinical trials have proposed a useful effect of helminth infections on improvement of inflammatory disorders. In this study, we investigated the immunomodulatory effect of Syphacia obvelata infection to induce intestinal tolerance in C57BL/6 mice. Mice were infected through the cagemates with self-infected BALB/c mice. Four weeks post-infection, expression levels of IFN-γ, TNF-α, IL-17, and IL-10 were assessed in the supernatant of mesenteric lymph node (MLN) culture. Foxp3⁺Treg were measured in MLN cells by flow cytometry. In the S. obvelata-infected group, the percentage of Tregs (5.2±0.4) was significantly higher than the control (3.6±0.5) (P<0.05). The levels of IL-10 (55.3±2.2 vs 35.2±3.2), IL-17 (52.9±3.8 vs 41±1.8), IFN-γ (44.8±4.8 vs 22.3±2.3) and TNF-α (71.1±5.8 vs 60.1±3.3) were significantly increased in infected mice compared to the control group (P<0.05). The above results showed the potential effects of S. obvelata to induce intestinal tolerance. Therefore, it seems that S. obvelata may increase the immunological suppressive function in the intestinal tract.
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Animals , Mice , Flow Cytometry , Helminths , Hope , Immune System , Interleukin-10 , Interleukin-17 , Lymph Nodes , OxyuroideaABSTRACT
Objective To investigate whether there are correlation among changes of Treg/Th17 ratio with virological response and serological re?sponse in patients with hepatitis B e antigen(HBeAg)?positive chronic hepatitis B(CHB)after receiving entecavir. Methods 27 patients with HBeAg?positive CHB were enrolled for the study. Peripheral blood Treg cell frequency and Th17 cell frequency,HBV DNA,alanine aminotransfer?ase levels,hepatitis B virus marker were detected before and after entecavir treatment at 4,12,24,36,and 48 weeks. Peripheral blood Treg cell frequency and Th17 cell frequency of 20 healthy volunteers were detected as well. Results Treg/Th17 ratio of patients decreased from 4 week and reached the lowest point at 12 weeks. Treg/Th17 ratio(12 weeks)of the patients who got the early virological response and HBeAg disappeared was lower than others,and the difference was statistical significant. Conclusion Treg/Th17 ratio changes in HBeAg?positive CHB patients receiv?ing entecavir antiviral treatment. Treg/Th17 ratio(12 weeks)showed significant correlation with early virological response and HBeAg disappear?ance. The changes of Treg/Th17 ratio may provide a reference of curative effect.
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Objective To investigate the expression levels of forkhead transcription factor 3 (FOXP3) and cytotoxic T lymphocyte-associated antigen-4(CTLA-4) in the peripheral blood of children with newly diagnosed type 1 diabetes mellitus(T1DM),and explore their roles in the pathogenesis of T1DM. Methods Fifty children with newly diagnosed T1DM and 30 healthy children( control group) were select-ed. FOXP3 and CTLA-4 mRNA expressions were detected using real-time PCR. Soluble FOXP3 ( sFOXP3 ) and CTLA-4(sCTLA-4) protein levels in the serum were assayed by ELISA,and the levels of diabetes anti-bodies,HbA1C and C-peptide were determined using Western blotting,high performance liquid ion-exchange chromatography,and electrochemiluminescence, respectively. Results The expressions of FOXP3 mRNA and protein in T1DM patients were lower than those in the control group[0. 95 ± 0. 48 vs. 2. 11 ± 0. 79,(6. 27 ± 1. 49) ng/ml vs. (9. 02 ± 2. 37) ng/ml,respectively,both P <0. 01],while the expressions of CTLA-4 mRNA and protein were higher than those in the control group[2. 43 ± 0. 83 vs. 1. 94 ± 0. 84,(77. 88 ± 22. 34) ng/ml vs. (65. 97 ± 12. 11) ng/ml,respectively,both P<0. 01]. In T1DM patients,the expressions of FOXP3 and CTLA-4 genes presented positive correlation with the protein expression( r=0. 758,0. 396, P<0. 05), and FOXP3 protein expression presented correlation with CTLA-4 protein expression ( r =-0. 624,P<0. 05). Conclusion There are abnormal expressions of FOXP3 and CTLA-4 genes as well as their proteins in the peripheral blood of the children with newly diagnosed T1DM,and FOXP3 regulated the expression of CTLA-4 in T regulatory cells,indicating the involvement of immune mechanisms in the devel-opment of T1DM.
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Objective We aimed to evaluate the role of CD4 + CD25 + T regulatory cells in predicting the prognoses of patients with sepsis.Methods Patients with sepsis in Shanghai Changzheng Hospital from December 2013 to April 2014 were identified and grouped into survival group (SG,n =19) and death group (DG,n =9) in accordance with their clinical outcomes.CD4 + CD25 +T regulatory cell ratio,C-replication protein,bilirubin,procalcitonin,and coagulation function were tested on the 1st day and 7th day,and APACHE Ⅱ and SOFA were analyzed to assess the predictability of this group of cells.Results Twentyeight patients were identified,with a mean age of (60.36 ± 15.30) years,a mean APACHE Ⅱ score of (16.68±7.00),and a mean SOFA score of (7.18 ±3.78).Twelve (42.9％) of the individuals were accompanied with severe multiple trauma,and 10 (35.7％) were in septic shock,and 9 (32.2％) died of severe sepsis.The first day CD4 + CD25 + T regulatory cell ratios on the first day were 2.10％ (0.80,3.10)％ (SG) vs.1.80％ (1.15,3.65)％ (DG) (Z=-0.148,P=0.883),andonday7 were 0.90％ (0.30,2.80)％ (SG) vs.5.70％ (2.60,8.30)％ (DG) (Z=-2.905,P=0.004) presented significant predictability.Conclusions Dynamic monitoring of CD4 + CD25 + T regulatory cells could predict the prognoses of patients with sepsis and should be generalized in clinical emergency practice.
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Objective To explore the expression characteristic of T regulatory cell (Treg)/T help cell (Th)17 and the correlation to disease progression in chronic hepatitis B (CHB) patients undergoing antivirus treatment,and to explore their roles in pathogenesis of CHB.Methods A total of 53 patients with CHB (CHB group) and 21 healthy controls (healthy control group) were selected,CHB patients included mild 18 cases,moderate 16 cases and severe 19 cases.The expression of Treg and Th17 were detected by flow cytometry and compared.The levels of alanine aminotransferase (ALT),total bilirubin (TBIL),cholinesterase,albumin were measured by automatic biochemical machine and for correlation analysis.Results The levels ofTh17,Treg,Treg/Th17 were (2.13 ± 0.65)％,(2.99 ± 0.68)％ and (6.07 ± 1.18)％,(5.14 ± 0.96)％ and 2.86 ± 0.67,1.73 ± 0.45 in CHB group before and after treatment for 24 weeks and (3.59 ± 0.75)％,(4.02 ± 0.77)％,1.04 ± 0.34 in healthy control group.There were significant differences (F =14.78,10.12,17.19; P ＜ 0.01).The level of Th17 in mild,moderate and severe CHB was gradually decreased,but there was no significant difference (F =1.10,P =0.337).The level of Treg in mild,moderate and severe CHB patients was gradually increased,but there was no significant difference (F =0.54,P =0.585).The level of Treg/Th17 was gradually increased with aggravation (2.58 ± 0.59,2.76 ± 0.63,3.21 ± 0.71),and there was significant difference (F =3.15,P ＜ 0.01),the level of Treg/Th17 in severe CHB patients was significantly higher than that in mild,moderate CHB patients (P ＜0.05).The level of Treg/Th17 had significantly positive correlation with ALT,TBIL (r =0.272,P=0.000; r =0.226,P=0.000).Conclusion Treg/Th17 balance not only relates with the pathogenesis of CHB,but also with the related immune inflammatory of liver tissue.
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The aim of the study is to characterize the phenotypes of CD4+ CD25+ T regulatory cells within the liver granulomas and association with both Foxp-3 gene expression and splenic cytokines. Naive C57BL/6 mice were intravenously injected with multiple doses of the soluble egg antigen (SEA) 7 days before cercarial infection. The immunized and infected control groups were sacrificed 8 and 16 weeks post-infection (PI). Histopathology, parasitological parameters, splenic phenotypes for T regulatory cells, the FOXP-3 expression in hepatic granuloma using real-time PCR, and the associated splenic cytokines were studied. Histopathological examination of the liver revealed remarkable increase in degenerated ova within hepatic granuloma which decreased in diameter at weeks 8 and 16 PI (P<0.01). The percentage of T regulatory cells (CD4+ CD25+) increased significantly (P<0.01) in the immunized group compared to the infected control at weeks 8 and 16 PI. The FOXP-3 expression in hepatic granulomas increased from 10 at week 8 to 30 fold at week 16 PI in the infected control group. However, its expression in the immunized group showed an increase from 30 at week 8 to 70 fold at week 16 PI. The splenic cytokine levels of pro-inflammatory cytokines, IFN-gamma, IL-4, and TNF-alpha, showed significant decreases (P<0.05) compared to the infected control group. In conclusion, the magnitude and phenotype of the egg-induced effects on T helper responses were found to be controlled by a parallel response within the T regulatory population which provides protection in worm parasite-induced immunopathology.
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Animals , Humans , Mice , Antibodies, Helminth/immunology , Antigens, Helminth/administration & dosage , Cytokines/genetics , Forkhead Transcription Factors/genetics , Granuloma/immunology , Immunization , Mice, Inbred BALB C , Schistosoma mansoni/genetics , Schistosomiasis mansoni/genetics , Spleen/immunology , T-Lymphocytes, Regulatory/immunologyABSTRACT
Help T cell 1/Help T cell 2 and some cytokines disequilibrium can give a suitable explanation for hypersensitivity and hypogammaglobulinemia in primary nephritic syndrome(PNS) patients. The disturbance of regulatory T cell(Treg cell) and Th17 cell can lead to correlated cytokines derangement, which explained the pathogenesis of PNS from another aspect. Refractory nephrotic syndrome can be effectively treated by rituximab followed the percentage of regulatory T cell increasing, which indicated that Treg may play an important role in pathogenesis of PNS.
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Objective:To investigate the expression of a new cytokine,thymic stromal lymphopoietin (TSLP) and its receptor TSLPR in the villi of human first trimester gestation.Methods:Villi were collected from women who had undergone an artificial abortion at 7-11 weeks of normal gestation.The trophoblast cells (Tros) were isolated and cultured.The total RNA was extracted using TRIzol reagent,from both villi and the Percoll-gradient-isolated Tros,then the DNA fragments of hTSLP and hTSLPR were amplificated by RT-PCR.Villous tissues were detected for TSLP by immunohistochemistry (IHC) and Western blot.Immunocytochemistry (ICC) was carried out on cultured trophoblast cells for TSLP/TSLPR expression.Levels of TSLP in the supernatants were detected by ELISA.Results:Normal villi and the cultured Tros transcript were found to express TSLP/TSLPR mRNA and secreted TSLP protein.In addition,TSLP receptor was also expressed on trophoblasts.Conclusion:Both TSLP and its receptor are expressed on villi and trophoblast cells,which suggests that TSLP plays an important role in maternal-fetal immuno-tolerance in human early pregnancy.