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Objective:To investigate whether anisodamine can regulate the ratio of helper T helper cells/regulatory T cells (Th17/Treg) and its protective effect on animals after resuscitation.Methods:Twenty-four Beijing white minipigs were randomly divided into sham operation group (Sham group), resuscitation and normal saline group (SA group), and resuscitation and anisodamine hydrobromide group (AH group), with 8 pigs in each group. In SA group and AH group, ventricular fibrillation was induced by continuous stimulation with intraventricular electrodes for 8 minutes and then resuscitated to establish ischemia/reperfusion (I/R) model. In SA group, after cardiopulmonary resuscitation (CPR), only normal saline was intravenously infused, while in AH group, normal saline and anisodamine hydrobromide were given intravenously at the same time point. Hemodynamic indexes, arterial blood gas analysis indexes, interleukins (IL-17, IL-10) levels in venous blood and IL-17/IL-10 ratio were recorded at 6 different time points: baseline, immediately after return of spontaneous circulation (ROSC), 1 hour, 2 hours, 4 hours and 6 hours after ROSC. The animals were sacrificed at 6 hours after ROSC, and intestinal lymphatic tissues were taken to observe pathological changes under light microscope. At the same time, the levels of IL-17 and IL-10 in intestinal lymphatic tissue were measured (the ratio of IL-17/IL-10 represents the ratio of Th17/Treg cytokines) to evaluate the immune status of the resuscitated animals. The bacterial translocations of different groups were evaluated by culturing intestinal lymphoid tissue.Results:With the extension of ROSC time, the levels of IL-17 in venous blood and the IL-17/IL-10 ratio in pig blood samples continued to decrease, while the levels of IL-10 continued to increase. From 2 hours after ROSC, the IL-17/IL-10 ratio in AH group was significantly higher than that in SA group continued until at 6 hours after ROSC (0.79±0.05 vs. 0.49±0.08, P < 0.05). Light microscopy showed that the number and size of lymph nodules in the cortex of intestinal lymphatic tissue were less in AH group, compared with SA group. Compared with Sham group, the levels of IL-17 and IL-17/IL-10 ratio also decreased in intestinal lymphatic tissue at 6 hours after ROSC [IL-17 (ng/L): 155.23±0.92, 178.76±7.25 vs. 209.21±19.82, IL-17/IL-10 ratio: 1.43±0.13, 1.92±0.18 vs. 3.30±0.31, all P < 0.05], and IL-10 increased significantly (ng/L: 109.85±11.60, 93.55±81.83 vs. 63.45±0.62, all P < 0.05); IL-17/IL-10 ratio in AH group was significantly higher than that in SA group (1.92±0.18 vs. 1.43±0.13, P < 0.05). Tissue culture indicated the intestinal bacterial translocation after resuscitation, suggesting that the animals had immunosuppression and the increased risk of intestinal secondary infection after resuscitation. Compared with SA group, the risk of bacterial translocation was lower than that in AH group [62.5% (5/8) vs. 87.5% (7/8), P < 0.05]. Conclusions:Anisodamine plays an immunomodulatory role by affecting the balance of Th17/Treg cytokines in resuscitated animals, so as to reduce the risk of intestinal secondary infection and has an organ protective effect.
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【Objective】 To evaluate the effect of Arntl on T cell development and T cell-mediated anti-viral immunity. 【Methods】 ArntlF/FCD4cre+(KO) in mice was constructed to delete Arntl gene specifically in T cells. We examined the percentage and number of T cell subsets in the thymus and spleen by flow cytometry (FCM). At day 8 after lymphocytic choriomeningitis virus (LCMV) infection, the proportions of T cell subsets, virus-specific CD8+ T cells and IFN-γ secreting T cells were analyzed. The viral load in the spleen was measured using qPCR. Naive CD4+ T cells (CD4+CD25-CD44-CD62L+) were sorted by flow cytometry to perform T helper cell differentiation in vitro. 【Results】 The percentage and number of T cells in the thymus and spleen of KO mice showed no significant change compared with those in the control group (ArntlF/FCD4cre- mice, WT) (P>0.05). Acute LCMV infection did not cause observable changes in effector T cell proportion in the spleen of KO mice compared to that in WT mice (P>0.05), but KO mice showed a higher proportion of IFN-γ secreting T cells (P<0.05) and better virus clearance (P<0.05). In addition, naive CD4+ T cells from KO mice were more prone to differentiate into Th1 cells in vitro (P<0.05). 【Conclusion】 Arntl deletion in T cells does not affect T cell development, but enhances their ability to defend against viral infection by promoting Th1 cell differentiation and response.
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The pathogenesis of psoriasis is complex,including genetic,immune and many other factors,and Th cell play an important role in this process. Vitamin D excerts excellent therapeutic effect on psoriasis,and the therapeutic effect may be related to T cells. In this passage,we look into the recent advances in the pathogenesis of psoriasis from the perspectives of Th1/Th2,Th9,Th17,Treg and the advances in treatment of psoriasis with vitamin D,for providing some ideas and entry points for future research.
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Objective To compare the effect of compound flumethasone ointment and clobetasol propionate cream on serum and skin lesion Th cell related indicators in patients with eczema. Methods 156 patients with chronic eczema were chosen. According to the type of topical drugs, they were divided into two groups: the flumethasone group and the clobetasol propionate group. The changes of eczema treatment effect, serum and skin lesions Th cell related indicators of the two groups were compared. Results After treatment, the serum interferon-γ (IFN-γ) of the flumethasone group was (27.57 ± 5.67) pg/mL, IL-2 was (36.51 ± 8.03) pg/mL and IL-4 was (26.37 ± 5.29) pg/mL, IL-10 was (25.38 ± 4.64) pg/mL and INF-γof skin lesions was (56.53 ± 21.81) pg/L , IL-2 was (51.69 ± 15.67) pg/L, IL-4 was (159.42 ± 25.64) pg/L and (139.62 ± 24.58) pg/L, significantly lower than those of clobetasol propionate group (P <0.05), but the clinical benefit rate (94.87%) was significantly higher than (80.77%) of clobetasol propionate group (P <0.05). Conclusion Compared with clobetasol propionate cream, the effect of compound flumetasone ointment is more effective in treating eczema, and its mechanism may regulate the expressions of Th cell related cytokines.
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Objective To explore the effect of 1-alpha,25-dihydroxy-cholecalcifero(1,25(OH)2D3)on liver fibrosis and its mechanism. Methods Degree of liver fibrosis was assessed through pathological detection and blood biochemical examination of liver function. Immunohistochemical assay was used to detect expressions of α-SMA,TGF-βand collagen I to observe activation level of hepatic stellate cells. Impact of 1,25(OH)2D3 on CD4+T cell differentiation was analyzed by flow cytometry,ELISA,and RT-PCR. Results 1,25(OH)2D3 improved the structure of the liver tissue and liver fibrosis. Expressions of collagen I ,TGF-βandα-SMA were significantly ele-vated in the liver tissue in rats with fibrosis(P 0.01);and ex-pressions of RORγt and T-bet decreased whereas GATA3 expression increased(P>0.01);as compared with those in the control group. Conclusions 1,25(OH)2D3 can alleviate the degree of liver tissue by lowering HSC activation and regulating Th cell differentiation.
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Objective To evaluate the effect of azitromycin upon the bronchiolitis obliterans and T helper (Th)17/regulatory T cell (Treg) balance after lung transplantation. Methods Twenty-four specific pathogen free(SPF) C57BL/6 mice were used as the donors and 48 Balb/c mice were utilized as the recipients. The Balb/c mice were randomly divided into the control (C group), azitromycin control (Cazm group), transplantation (T group) and transplantation + azitromycin groups (Tazm group), 12 mice in each group. In the T and Tazm groups, heterotopic tracheal transplantation models were established to simulate bronchiolitis obliterans after lung transplantation. From 1 d post-transplantation, intragastric administration of azitromycin was given at a dose of 30 mg/kg three times per week in the Cazm and Tazm groups. At 14 and 28 d after transplantation, the transplanted trachea was removed and peripheral blood was collected. The tracheal sample was prepared for hematoxylin-eosin (HE) staining for pathological observation. The expression levels of ROR-γt and Foxp3 messenger ribonucleic acid (mRNA) in the peripheral blood were quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). The variation in the related cytokines levels of Th17 cells and Treg in the plasma was detected by enzyme linked immunosorbent assay (ELISA). Results After heterotopic tracheal transplantation, compared with the C group, thetracheal occlusion accompanied with inflammatory infiltration was observed in the T and Tazm groups. The severity of relevant symptoms in the Tazm group was slighter than that in the T group. Compared with the T group, the expression level of ROR-γt mRNA in the Tazm group was significantly down-regulated (P<0.05). No statistical significance was identified in the expression of Foxp3 mRNA between two groups (P>0.05). Compared with the T group, the levels of interleukin (IL)-6 and IL-17 cytokines in the Tazm group were significantly down-regulated (all P<0.05). Conclusions Persistent therapy of azitromycin can delay the progression of bronchiolitis obliterans after transplantation, which is probably associated with inhibiting Th17 cell differentiation and inflammation.
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Objective To determine the protein and mRNA levels ofinterleukin-17 (IL-17) and the proportion of Th17 cells in the peripheral blood of patients with systemic lupus erythematosus (SLE) and their clinical significance is analyzed. Methods Twenty-five hospitalized SLE patients were recruited and twentytwo healthy volunteers were enrolled as normal controls. Plasma protein and mRNA of IL-17 in the peripheral blood were measured by enzyme-linked immunosorbent assay and real time-PCR respectively. Flow cytometric assay was used to analyze the percentage of Th17 cells in SLE patients. The relationship between IL-17/Th17 cells and clinical or laboratory parameters of SLE patients was explored. Students' t-test and Spearman's correlation was used to evaluate the relationship between mRNA level and inflammatory parameters. Results The plasma concentration and mRNA level of IL-17 was significantly elevated in SLE patients as compared to the normal controls (P＜0.05). The percentage of Th17 cells in patients with SLE was higher than that of normal controls and was significantly increased in more active SLE patients and SLE with nephritis than less active SLE and SLE without nephritis (P＜0.05). Both plasma levels of IL-17 and the proportion of Th17 cells were positively correlated with SLEDAI (r=0.681, P＜0.01; r=0.426, P=0.034). Conclusion Plasma IL-17 protein and mRNA expression level and the percentage of Th17 cells in SLE patients are all significantly elevated and the close relationship between IL-17/Th17 cells and disease activity suggests that IL-17/Th17 may play an important role in the pathogenesis of SLE.
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Objective To investigate the role of PPAR-γ in the gene expression of T-bet/GATA-3 in Jurkat T cells,and to explore the mechanisms underling this sensitizing effect of the change of TH cell subpopulation group.Methods Jurkat T cells were stimulated with PPAR-γ agonist pioglitazone.TH cell related cytokine IFN-γ and IL-10 was detected by ELISA,and the expression of transcription factors(T-bet and GATA-3)mRNA was detected by RT-PCR.To prove the PPAR-γ-dependent effect.the PPAR-γ-specific antagonist GW9662 was used.Results Stimulated with agonist PPAR-γpioglitazone.the concentration of IFN-γ and IL-10 and the expression of transcription factor T-bet and GATA-3 mRNA were both significantlY decreased in Jurkat T cells obviously,and these actions were dependent on the time and the concentrations of pioglitazone.Added with antagonist GW9662 at the same time,such inhibitory actions of IFN-γ and T-bet expression were recovered.but not IL-10 and GATA-3.Conclusion Pioglitazone can inhibite T cells proliferation and their secretion of cytokines.Pioglitazone can inhibit TH1 cells from secreting cytokines,and it is a PPAR-γ-dependent effect related to T-bet.The inhibition on TH2 is not a PPAR-γ-dependent effect and it is GATA-3 related.
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O objetivo dessa tese foi avaliar a expressão de citocinas Th1 (IL-12 e INFγ), citocinas Th2 (IL-4, IL-6 e IL-10) e das citocinas pró-inflamatórias IL-18, IL-1β e TNFα no fluido gengival de pacientes com periodontite crônica portadores da doença de Crohn (DC), de retocolite ulcerativa idiopática (RCUI) e em indivíduos saudáveis (o grupo controle, GC). Como objetivo secundário, avaliamos a função dos neutrófilos no fluido gengival desses pacientes através da mensuração das metaloproteinases da matriz -8, -9 (MMP-8 e MMP-9) e da atividade da elastase. Quinze pacientes com DC (idade média 38.2 ± 11.4 anos), 15 pacientes com RCUI (idade média 45.0 ± 10.5 anos) e 15 pacientes saudáveis (idade média 42.1 ± 7.8 anos) participaram desse estudo. Todos os dentes presentes, com exceção dos terceiros molares, foram examinados. Profundidade de bolsa (PB), nível de inserção clínica (NI), presença de placa e de sangramento a sondagem foram avaliados em seis sítios por dente. Em cada paciente, o fluido de 4 sítios com periodontite (PB ≥ 5 mm e NI ≥ 3mm) e de 4 sítios com gengivite (PB ≤ 3 mm e NI ≤ 1 mm) foram coletados através de pontas de papel absorvente pré-fabricadas. O sistema LUMINEX® foi utilizado na mensuração das IL-1β, IL-4, IL-6, IL-10, IL-12p70, TNFα, INFγ, MMP-8 e MMP-9. A IL-18 foi analisada através do ensaio ELISA e a atividade de elastase através de uma reação enzimática. O soro desses pacientes também foi analisado e o coeficiente de correlação de Pearson foi utilizado na análise da correlação entre as citocinas no soro e no fluido gengival. Nos sítios com gengivite, a quantidade total de IL-4 foi significativamente menor no grupo RCUI do que no grupo GC (p=0.016). Nos sítios com periodontite, a quantidade total de IL-4 foi significativamente menor no grupo DC do que no grupo GC (p=0.029)...
The aim of this thesis was to evaluate the expression of Th1 cytokines (IL-12 and INF-γ), Th2 cytokines (IL-4, IL-6 and IL-10) and the pro-inflammatory cytokines IL-18, IL-1β and TNF-α in the gingival crevicular fluid (GCF) from Crohns disease (CD) patients, ulcerative colitis (UC) patients and healthy individuals (control group, CG) who had chronic periodontitis. Besides, we measured elastase activity, matrix metalloproteinase -8 and -9 (MMP-8 and -9) to address the neutrophil function in the GCF. Fifteen CD patients (mean age 38.2 ± 11.4 years), 15 UC patients (mean age 45.0 ± 10.5 years) and 15 systemically healthy controls (mean age 42.1 ± 7.8 years) were enrolled in this study. All the present teeth, except for the third molars were examined. Probing pocket depth (PPD), clinical attachment loss (CAL), presence of plaque and presence of bleeding on probing were assessed in six sites per tooth. In every subject, GCF from 4 gingivitis sites (PPD ≤ 3mm and CAL ≤ 1mm) and from 4 periodontitis sites (PPD ≥ 5mm and CAL ≥ 3mm) were collected with filter strips. The data were reported as total amount and concentration. IL-1β, IL-4, IL-6, IL-10, IL-12p70, TNFα, INFγ, MMP-8 and MMP-9 were analyzed by the Luminex® analyzer. IL-18 was analyzed using a commercially available ELISA assay and the elastase activity by an enzymatic reaction. The serum was also analysed and the correlations between the cytokines in the GCF and in the serum were calculated by Pearson correlation analysis. In gingivitis sites, the total amount of IL-4 was significantly lower in the UC group than in the CG group (p=0.016). In periodontitis sites, the total amount of IL-4 was significantly lower in CD group than in the CG group (p=0.029). The total amount of IL-4 was lower in UC group than in CD group (p=0.077)...
Subject(s)
Humans , Cytokines/chemistry , Cytokinesis/immunology , Gingival Crevicular Fluid/chemistry , Lymphocytes/chemistry , Chronic Periodontitis/enzymology , Case-Control Studies , Crohn Disease , Inflammatory Bowel Diseases , Leukocyte Elastase , Matrix Metalloproteinases/chemistry , ProctocolitisABSTRACT
BACKGROUND: The statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, are approved for cholesterol reduction, and may also be beneficial in the treatment of inflammatory disease. In this study, atorvastatin was tested in experimental colitis, a disease model of inflammatory bowel disease. METHODS: To induce colitis, dextran sodium sulfate (DSS) or trinitrobenzene sulfonic acid (TNBS) were administrated to C57BL/6 or BALB/c mice. Mice were monitored daily for loss of body weight and survival for indicated days. Colon length and histology were examined after sacrifice. RESULTS: The administration of DSS induced marked colonic inflammation and shortening, and resulted in a loss of body weight. DSSinduced colitis was not affected by atorvastatin treatment, but in contrast, the administration of atorvastatin relieved TNBS-induced colitis with a resultant rapid recovery of weight loss and a reduction in colonic length shortening. Histologically, inflammatory cell infiltration in the colonic wall, mucosal ulceration and crypt disruption were also suppressed in atorvastatin treated mice. CONCLUSION: These results suggest that atorvastatin preserves intestinal integrity in colitis, probably via the modulation of Th cell-mediated immune response, in a manner independent of innate immunity.
Subject(s)
Animals , Mice , Body Weight , Cholesterol , Coenzyme A , Colitis , Colon , Dextrans , Hydroxymethylglutaryl-CoA Reductase Inhibitors , Immunity, Innate , Inflammation , Inflammatory Bowel Diseases , Oxidoreductases , Sodium , Ulcer , Weight Loss , AtorvastatinABSTRACT
Objective:To study the gene expression spectrum of Th cell immune response in acute and chronic toxoplasma infection in BALB/c mice.Methods:BALB/c mice were infected with toxoplasma tachyzoite (1?104/m,0.5 ml) by intraperitoneal injection.Two mice were killed at 6th and 12th day postinjection.The immune response gene expressing levels in mice spleen cells were detected with 128 kind gene microarray chips.Partial of the results were determined with Real time-PCR.Results:There were 20 genes changed after toxoplasma infection.Eight of them (Gata-3,Ccr3,Ccr4,Bcl6,Nfatc1,CD80,Fos,CD69) were regulated upward and twelve genes (T-bet,CIITA,Irf1,Mapk9,Nfatc3,Fasl,Tyk2,Lat,Mapk10,Socs3,Socs6 and Yy1) were regulated downward.On the whole,expression of Th2 response genes presented a up trend,and Th1 response genes presented a down trend.The results of Real time-PCR were accordant with microarray in the expression of Gata-3 and T-bet gene.Conclusion:According to the Th cell immune response gene expression spectrum,Th1 immune response is dominant in the acute infention when mice are infected by toxoplasma.Then,it is biased toward Th2 immune response.
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Objective To observe the apoptosis of Th1 and Th2 cells in C57BL/6 mice chronically infected with Schistosoma japonicum.Methods The apoptotic Th1 and Th2 cells in spleen and lymph node from C57BL/6 mice infected with Schistosoma japonicum for 13 weeks were examined by three-color and indirect flow cytometery with staining surface molecule and intracellular cytokines.Results Compared with the normal mice,the proportion of apoptotic Th1 and Th2 cells of 13-week post-infection was significantly high,and the apoptotic Th1 cells increased more than apoptotic Th2 cells in the infected C57BL/6 mice,and the Th1 cells were more susceptible to apoptosis than Th2 cells.Conclusions Unequal susceptibility to apoptosis in Th1 and Th2 cells may be one of the reasons leading to Th2 polarization on mice chronically infected with Schistosoma japonicum,which provides the new proof of Th polarization.
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Objective:To investigate the changes of Th cell differentiation in the VMC and interference effect of Qingxin capsule on them.Methods:BALB/c mouses with different courses of VMC were established, after treated with Qingxin capsule, the cardiac pathological changes were observed by light microscope and transmisson electron microscope,levels of IL-2,IL-4,IL-10 and IFN-? in blood serum were detected by ELISA.Results: Whatever in acute stage or recovery stage of VMC, the myocard pathological changes of mouses were lighter after treated with Qingxin capsule;moreover, levels of IFN-? and IL-2 in blood serum of mouses with VMC decreased significantly(P0.05).Conclusion:Qingxin capsule can restore the balance of Th1 and Th2 cells through inhibiting reaction of Th1 and enhancing reaction of Th2.