Peripheral lymphocytes and intracellular cytokines in C282Y homozygous hemochromatosis patients

BACKGROUND/AIMS: Several abnormalities in the immune status of hereditary hemochromatosis patients have been reported. We evaluated the peripheral blood lymphocytes phenotype and cytokine profile of CD8(+) and CD4(+) T cells in C282Y homozygous hereditary hemochromatosis patients compared to control subjects. METHODS: Peripheral blood lymphocytes from 17 asymptomatic patients and 14 control subjects were analyzed. We determined the distribution of lymphocyte subsets and investigated at single-cell level by flow-cytometry the potential of cytokines production. The frequency of cytokine (interferon gamma, tumor necrosis factor alpha, interleukin 2 (IL-2), IL-4, IL-5, IL-10 and IL-13) producing cells was assessed in total T-lymphocytes, CD3(+)CD8(+) and CD3(+)CD4(+) subsets. RESULTS: The patients studied showed a significant decrease of total lymphocyte count, T CD4(+)CD3(+), CD28(+), CD8(+)CD28(+) lymphocytes and natural killer (NK) CD56(+)CD16(+)CD3(-) cells. The reduction of CD28(+) and CD8(+)CD28(+) lymphocyte count was inversely related to transferrin saturation index. An increase in the ability of T-cells to produce all the cytokines studied and a major increase in IL-4 and IL-10 production in the CD3(+)CD8(+) subset was found. Our results demonstrate that activated Th1 and Th2 lymphocytes coexist in the peripheral blood of hereditary hemochromatosis patients and that T-cytotoxic (Tc) 2 subset is more expanded than in control population. CONCLUSIONS: The association of a decreased number of T CD8(+) cytotoxic lymphocytes and NK cells, and the development of Tc2 cells in asymptomatic C282Y homozygous patients represents an imbalance in their immune function that might contribute to the high incidence of hepatocarcinoma.     

外周血Th17细胞及IL-17^＋FoxP3^＋T细胞在非小细胞肺癌患者的表达变化

目的：探讨辅助性T17（Th17）细胞及白细胞介素17（IL-17）＋叉头蛋白3（FoxP3）＋T细胞在非小细胞肺癌患者外周血中的表达及意义。方法：57例非小细胞肺癌患者作为研究对象．25名正常人作为对照。采用流式细胞术检测外周血Th17细胞、调节性T细胞（Treg细胞）的百分率以及IL-17＋FoxP3＋T细胞占Treg细胞的百分率。结果：Ⅳ期非小细胞肺癌患者外周血单核细胞（PBMC）中Th17细胞百分率高于Ⅰ-Ⅲ期患者及正常对照（均P〈0．01）。非小细胞肺癌患者PBMC中的Treg细胞百分率高于正常对照,并且IL-17＋FoxP3＋T细胞占Treg细胞的百分率高于正常对照（P〈0．05）。结论：Ⅳ期非小细胞肺癌患者中Th17细胞以及II-17＋FoxP3＋T细胞数量增加,Th17以及IL．17＋FoxP3＋T细胞可能参与了非小细胞肺癌的肿瘤远处转移过程．从而影响肿瘤进程。     

Increase in CD95 (Fas/APO-1)-positive CD4+ and CD8+ T cells in peripheral blood derived from patients with autoimmune hepatitis or chronic hepatitis C with autoimmune phenomena

BACKGROUND: The expressions of CD95 (Fas/APO-1) and Bcl-2 are determinants of apoptosis in normal lymphocytes, and abnormalities in their expressions might contribute to the induction of autoimmunity. In this study, we examined the expressions of CD95 and Bcl-2 on freshly isolated T and B cells from patients with autoimmune hepatitis (AIH) or chronic hepatitis C associated with autoimmune phenomena (CH-C(AI)). METHODS: The CD95 and Bcl-2 expressions within CD4+ T, CD8+ T, and CD19+ B cell subsets were analysed by two-colour flow cytometry. RESULTS: The surface expression of CD95 was significantly high in both the CD4+ T and CD8+ T cell subsets derived from the patients with AIH and those with CH-C(AI), compared with expression in patients with CH-C and normal subjects. The increase in CD95 expression was associated with the phenotypic conversion of naive CD45RO- to primed CD45RO+ CD4+ T cells. Bcl-2 was detected in the vast majority of peripheral T and B cells. There was no significant difference in the percentage of Bcl-2-positive cells in the CD4+ T cell, CD8+ T cell and CD19+ B cell subsets among the patient groups and normal subjects. CONCLUSIONS: These results indicate that an increase in CD4+ T cells expressing CD45RO and CD95 marks an important subset of AIH and CH-C(AI) patients. These expanded CD95+ CD45RO+ primed T cells most likely reflect a continuous antigen-specific or non-specific activation of T lymphocytes, and/or the persistent presence of activated lymphocytes as a consequence of abnormalities in the peripheral deletion of activated lymphocytes. These persistently activated lymphocytes might play a role in the induction of autoimmunity in AIH and CH-C(AI).     

结核性胸膜炎患者胸液中CD4~+ CD25~+ FoxP3~+调节T细胞抑制抗结核免疫

目的研究结核性胸膜炎患者胸液中CD4+CD25+FoxP3+调节T细胞是否增多,这些调节T细胞是否抑制结核的特异细胞免疫反应。方法使用细胞分离、流式细胞分析及体外细胞培养作细胞增殖及增殖抑制等实验方法,对15例结核性胸膜炎患者及17例健康正常人群胸液及外周血白细胞中CD4+CD25+FoxP3+调节T细胞的量及特征作研究。结果结核性胸膜炎患者胸液中CD4+CD25+FoxP3+调节T细胞明显高于患者及健康人群外周血。在体外,结核性胸膜炎患者胸液中单核细胞对BCG刺激产生γ-干扰素(IFN-γ)的能力明显强于患者及健康人群外周血中单核细胞;把这些调节T细胞从胸液单核细胞中清除,增强了结核患者胸液单核细胞对BCG刺激产生IFN-γ;从结核患者胸液分离的这些调节T细胞能抑制结核患者Th1细胞产生IFN-γ。结论结核性胸膜炎患者胸液CD4+CD25+FoxP3+调节T细胞增多,抑制结核性胸膜炎患者Th1细胞免疫反应,从而参与了结核性胸膜炎的发病。     

Asthma severity is associated with an increase in both blood CXCR3+ and CCR4+ T cells

OBJECTIVES: A predominance of type 2 helper T cells (Th2) in the bronchoalveolar space and peripheral blood is a well-accepted feature of bronchial asthma. However, the relationship between peripheral blood Th2 cells and asthma severity has not been thoroughly investigated. METHODS: As Th1 cells predominantly express the chemokine receptor CXCR3 and Th2 cells express CCR4, we assessed the distribution of peripheral blood CXCR3+ and CCR4+ lymphocytes using flow cytometry in 186 patients with asthma and 75 normal subjects. RESULTS: The proportion of CXCR3+/CD45RO+ cells in CD4+ T cells increased as the severity of asthma increased. The percentage of CCR4+/CD45RO+ cells in CD4+ T cells were elevated in mild to severe asthma patients compared with controls. However, there was no significant difference in CCR4+/CD45RO+ cells between the mild to severe asthma patients. There was no relationship between the patient's age and the numbers of CXCR3+ or CCR4+ T cells. The percentage of CCR4+ cells in CD45RO+/CD4+ T cells correlated with the levels of total serum IgE (r = 0.630, P < 0.0001). CONCLUSIONS: The proportion of CCR4+ cells in blood memory helper T cells may be increased in patients with asthma and is associated with the level of serum IgE, but severity of asthma is also associated with the increase of blood CXCR3+ cells in memory helper T cells.     

Altered cytokine expression of peripheral blood lymphocytes in polymyositis and dermatomyositis

OBJECTIVE: To investigate the intracellular and soluble cytokine levels and T cell subsets in peripheral blood of patients with active and inactive polymyositis and dermatomyositis. METHODS: The frequencies of T and B lymphocytes, T helper (Th), and T cytotoxic (Tc) cells and of interferon gamma (IFNgamma), interleukin (IL)4, and IL10 expression of CD4+ or CD8+ cells were determined by flow cytometry. The concentrations of soluble cytokines were measured with commercial enzyme linked immunosorbent assays. RESULTS: In active dermatomyositis there was a decreased percentage of T (CD3+) lymphocytes and Tc (CD8+) lymphocytes, decreased IFNgamma expression of CD4+ and CD8+ cells, but an increase in B and IL4 producing CD4+ lymphocyte frequencies. These prominent changes disappeared in the inactive stage of the disease. In polymyositis no significant change in these lymphocyte subsets or in intracellular cytokine expression could be detected in either the active or the inactive form. The frequency of IL4+/IFNgamma+ Th cells was calculated and a significantly increased Th2/Th1 frequency was found in active dermatomyositis, and a decreased frequency in inactive dermatomyositis, compared with the control population. CONCLUSIONS: There appears to be a difference between polymyositis and dermatomyositis in the level of peripheral blood lymphocytes and their intracellular cytokine content. These findings provide further evidence for a difference in the pathogenesis of polymyositis and dermatomyositis.     

Expression of Th1 markers by lung accumulated T cells in pulmonary sarcoidosis

OBJECTIVES: The balance between Th1 and Th2 T cells, classified by virtue of their cytokine production can in an immune response influence the phenotype and progression of several clinical diseases. In this study, we examined the expression of Th1 associated chemokine and cytokine receptors CXCR3, CCR5, and interleukin (IL)-12R, IL-18R, respectively, as well as of the Th2 associated chemokine receptors CCR4 and CXCR4 on CD4+ and CD8+ T cells. SUBJECTS: Eighteen patients with untreated pulmonary sarcoidosis. MATERIALS AND METHODS: We used monoclonal antibodies and flow cytometry to analyse the expression of chemokine receptors CXCR3, CXCR4, CCR4 CCR5 and cytokine receptors IL-12R, IL-18R in combination with anti-CD4 and anti-CD8 mAbs in bronchoalveolar lavage fluid (BAL) and peripheral blood lymphocytes (PBL) from sarcoidosis patients. RESULTS: There were significantly more BAL CD4+ T cells expressing CXCR3, CCR5, IL-12R and IL-18R compared with paired PBL CD4+ T cells. In contrast, the Th2 associated chemokine receptors CXCR4 and CCR4 were expressed by a fewer percentage of BAL CD4+ compared with PBL CD4+ T cells. There was a positive correlation between the percentage of BAL lymphocytes and the number of CXCR3 and CCR5 expressing CD4+ BAL T cells. Also, the number of CD4+ IL-18R+ BAL fluid cells correlated negatively with disease duration. CONCLUSIONS: The lung accumulation of CXCR3, CCR5, IL-12R and IL-18R expressing T cells is in line with previous reports showing elevated levels in the lung of the corresponding ligands in sarcodosis. Blocking such ligands and/or receptors may develop into a future immunomodulatory therapy.     

Seasonal Variations of T-Cell Cytokine Pattern in Peripheral Blood from Atopic Subjects

Previous studies on the effect of seasonal exposure to the sensitizing antigen on T-cell cytokine pattern from atopic subjects evaluated T-cell cytokine production by titration in the serum or culture supernatants. The purpose of this study was to determine the seasonal variations of T-cell cytokine pattern from atopic subjects at the single-cell level. We examined the interleukin-4 (IL-4) and interferon -γ expression in peripheral blood CD4+ and CD8+ T cells from 11 subjects with grass-pollen-sensitive allergy before and during the 1999 grass pollen season using a flow cytometric method of intracellular cytokine detection. Eight healthy volunteers served as the control group. Flow cytometric analysis of peripheral blood lymphocytes showed no seasonal variations of IL-4- and interferon-γ-producing T cells in atopic subjects. However, there was a decreased percentage of IL-4-producing cells among peripheral blood CD4+ and CD8+ T cells from the atopic subjects both during and outside the pollen season in comparison to the controls. We did not find seasonal variations of T-cell cytokine pattern in peripheral blood from atopic subjects. However, we observed a decreased percentage of IL-4-producing T cells in peripheral blood from these subjects in comparison to healthy controls. These data add to the view of a continuous migration of T helper 2 (TH2) cells from the blood to the tissues of primary allergen exposure.     

Changes in expression of T-helper (Th) 1- and Th2-associated chemokine receptors on peripheral blood lymphocytes and plasma concentrations of their ligands, interferon-inducible protein-10 and thymus and activation-regulated chemokine, after antithyroid drug administration in hyperthyroid patients with Graves' disease

OBJECTIVE: Although Graves' disease is considered an autoantibody-mediated, T-helper 2 (Th2)-dominant disease, Th1-dominance may prevail in its initial phase. We longitudinally investigated Th1/Th2 balance in untreated hyperthyroid patients with Graves' disease after treatment of methimazole (MMI), an antithyroid drug. DESIGN: University clinic outpatients were studied prospectively. PATIENTS: Subjects included 23 untreated hyperthyroid patients with Graves' disease and 17 age-matched control subjects. METHODS: Before and after treatment, we measured Th1- and Th2-associated chemokine receptors (CXCR)3 and CCR4, on peripheral blood lymphocytes using flow cytometry, as well as plasma concentrations of their ligands, interferon-inducible protein (IP)-10 and thymus and activation-regulated chemokine (TARC). RESULTS: The percentage of CXCR3-expressing cells among CD4+T lymphocytes and plasma IP-10 was significantly higher in hyperthyroid Graves' disease patients than in controls. At 12 and 24 weeks after initiation of MMI, percentage of CXCR3-expressing CD4+T lymphocytes had decreased significantly, while the percentage of CCR4-expressing CD4+T lymphocytes had increased significantly at 24 weeks. The CXCR3/CCR4 ratio had decreased significantly at 24 weeks. Plasma concentrations of IP-10 had decreased significantly at 12 and 24 weeks. Plasma concentrations of TARC also had decreased significantly at 24 weeks. CONCLUSIONS: In hyperthyroid patients with Graves' disease in the active phase, Th1 cells rather than Th2 cells predominated among peripheral blood lymphocytes. After initiation of MMI, an ongoing transition from Th1 to Th2 dominance occurred.     

Major role for CD8+ T cells in the protection against Toxoplasma gondii following dendritic cell vaccination

Toxoplasma gondii is the causative agent of toxoplasmosis, a worldwide zoonosis for which an effective vaccine is needed. Vaccination with pulsed dendritic cells is very efficient but their use in a vaccination protocol is unconceivable. Nevertheless, unravelling the induced effector mechanisms is crucial to design new vaccine strategies. We vaccinated CBA/J mice with parasite extract-pulsed dendritic cells, challenged them with T. gondii cysts and carried out in vivo depletion of CD4⁺ or CD8⁺ T lymphocytes to study the subsequent cellular immune response and protective mechanisms. CD4⁺ lymphocytes were poorly implicated either in spleen and mesenteric lymph node (MLN) cytokine secretion or in mice protection. By contrast, the increasing number of intracerebral cysts and depletion of CD8⁺ cells were strongly correlated, revealing a prominent role for CD8⁺ lymphocytes in the protection of mice. Splenic CD8⁺ lymphocytes induce a strong Th1 response controlled by a Th2 response whereas CD8⁺ cells from MLNs inhibit both Th1 and Th2 responses. CD8⁺ cells are the main effectors following dendritic cell vaccination and Toxoplasma infection while CD4⁺ T cells only play a minor role. This contrasts with T. gondii infection which elicits the generation of CD4⁺ and CD8⁺ T cells that provide protective immunity.     

细胞免疫参数对结直肠癌的评估价值

目的:评估结直肠癌患者抗肿瘤免疫状态.方法:应用流式细胞术检测100例未转移结直肠癌患者、100例伴有转移结直肠癌患者和100例健康志愿者外周血CD4+、CD8+、NK、B、CD4+CD25HighCD127lowTreg、Th/Treg值,采用单因素方差分析进行比较,分析差异.结果:转移性结直肠癌患者组分别与正常对照组和非转移性结直肠癌患者组相比,CD4+CD25HighCD127lowTreg细胞升高(7.72%±2.20%vs6.08%±1.47%,5.91%±1.55%,均P<0.05),CD4+T细胞降低(34.04%±8.71%vs37.83%±7.62%,37.68%±8.89%,均P<0.05),Th/Treg值降低(4.70±1.72vs6.47±2.54,6.81±4.09,均P<0.05).结直肠癌患者与正常对照组CD8+T细胞、NK细胞、B细胞三组两两相比,均无统计学意义.结论:转移性结直肠癌患者免疫功能紊乱,主要表现为CD4+T细胞、Th/Treg值降低,CD4+CD25HighCD127lowTreg细胞升高.     

Pro- and anti-inflammatory cytokine production by autoimmune T cells against preproinsulin in HLA-DRB1*04, DQ8 Type 1 diabetes

Aims/hypothesis Preproinsulin is a target T cell autoantigen in human Type 1 diabetes. This study analyses the phenotype and epitope recognition of preproinsulin reactive T cells in subjects with a high genetic risk of diabetes [HLA-DRB1*04, DQ8 with Ab+ (autoantibody-positive) or without islet autoantibodies (control subjects)], and in HLA-matched diabetic patients.Methods A preproinsulin peptide library approach was used to screen for cytokine profiles and epitope specificities in human peripheral blood lymphocytes, and CD4⁺CD45RA^– and CD4⁺CD45RA⁺ T cell subfractions, representing memory and naive and recently primed T cells respectively.Results In CD4⁺ T cell subsets we identified immunodominant epitopes and cytokine production patterns that differed profoundly between patients, Ab+ subjects and non-diabetic HLA-matched control subjects. In Ab+ subjects, a C-peptide epitope C13–29 and insulin B-chain epitope B11–27 were preferentially recognised, whereas insulin-treated Type 1 diabetic patients reacted to native insulin and B-chain epitope B1–16. In peripheral blood lymphocytes of Ab+ subjects, an increase in T helper (Th) 1 (IFN, IL-2) and Th2 (IL-4) cytokines was detectable, wheras in CD45RA⁺ and CD45RA^– subsets, IL-4 and IL-10 phenotypes dominated, compatible with the contribution of non-CD4 cells to IFN content. In insulin-treated Type 1 diabetic patients, naive and recently primed CD4⁺ cells were characterised by increasd IFN, TNF, and IL-5.Conclusions/interpretation Our data show that T cell reactivity to preproinsulin in CD45RA subsets is Th2-dominant in Ab+ subjects, challenging the Th1 paradigm in Type 1 diabetes. Characteristic immunodominant epitopes and cytokine patterns distinguish diabetic patients and Ab+ subjects from HLA-matched healthy individuals. This could prove useful in monitoring of T-cell immunity in clinical diabetes intervention trials.Abbreviations PPI Preproinsulin - PBMC peripheral blood mononuclear cells - Th T helper cells - Ab+ autoantibody-positive - ICA islet cell antibodies - IA-2A islet thyrosine phosphatase - SI stimulation index - Tr T regulatory cells W. Karges and B.O. Boehm contributed equally to this article     

白介素23受体在支气管哮喘小鼠脾源性CD4+T细胞中的表达及意义

目的 探讨急性支气管哮喘(简称哮喘)小鼠CD4+T细胞表面白介素23受体(IL-23R)的表达及在哮喘发病中的作用.方法 建立急性哮喘小鼠模型,免疫磁珠分离小鼠脾源性CD4+T细胞,培养24 h后,检测CD4+T细胞表面IL-23R的表达、Th17细胞的阳性率及细胞培养上清液中的IL-17水平.结果 哮喘小鼠脾脏CD4+T细胞表面IL-23R的表达明显高于正常组(P＜0.01);哮喘小鼠脾脏CD4+T细胞中Th17细胞的阳性率明显高于正常组(P＜0.01);哮喘小鼠脾脏CD4+T细胞分泌IL-17浓度明显高于正常组(P＜0.01);小鼠脾脏CD4+T细胞表面IL-23R的表达与Th17细胞的阳性率和IL-17的浓度呈正相关.结论 IL-23R在急性哮喘的发病机制中可能起重要作用.     

Impairment of autologous mixed lymphocyte reaction in the spleen and peripheral blood lymphocytes of patients with idiopathic portal hypertension

The etiology of idiopathic portal hypertension (IPH) is unknown, although many studies have suggested that it might be an autoimmune disease. The autologous mixed lymphocyte reaction (AMLR) involves the proliferation of T lymphocytes when co-cultured with autologous non-T cells and may reflect immune control mechanisms in vivo. The AMLRs in the spleen and peripheral blood of three patients with IPH were measured and it was shown that the AMLRs both in the spleen and peripheral blood were significantly suppressed compared to those of normal healthy subjects. By allogeneic MLR, there was a tendency that the disturbance of non-T cells was more intensive than that of T cells. The AMLR of peripheral blood did not improve by splenectomy. Thus, the depressed cause of AMLR in patients with IPH was suggested mainly to disturbance of the antigen-presenting ability of non-T cells, and it was suggested that not only the spleen cells, but systemic immune disturbance caused the impairment of AMLR in IPH.     

Increased frequency of interferon-gamma-producing peripheral blood CD4+ T cells in chronic hepatitis C virus infection

OBJECTIVES: To determine the profile of cytokine secretion by CD4+ T helper (Th) cells in chronic hepatitis C virus (HCV) infection, we used flow cytometry to determine the percentage of interferon (IFN)-gamma and interleukin (IL)-4 producing cells from CD4+ T lymphocytes in peripheral blood obtained from patients chronically infected with HCV. METHODS: Peripheral blood mononuclear cells isolated from 89 HCV infected subjects (22 asymptomatic carriers, 56 patients with chronic hepatitis, and 11 patients with liver cirrhosis) and 24 healthy controls were stained with surface CD4 and intracellular IFN-gamma and IL-4. Serum soluble IL-2 receptor (sIL-2R) levels were analyzed by ELISA. RESULTS: The frequency of IFN-gamma producing CD4+ cells in asymptomatic HCV carriers, patients with chronic hepatitis, and patients with liver cirrhosis were significantly higher than those of healthy controls (p<0.01, respectively). In contrast, the percentages of IL-4-producing CD4+ cells were very low, and there were no significant correlations with disease progression. A significant elevation in serum sIL-2R levels was found in chronic HCV infection compared to healthy controls, and serum sIL-2R levels significantly correlated with the frequency of IFN-gamma-producing cells. CONCLUSIONS: In HCV infected subjects, both serum sIL-2R and IFN-gamma are increased in chronic HCV infection no matter the stage of disease, meaning they are no different in asymptomatic carriers, patients with chronic hepatitis, and patients with liver cirrhosis, and that Th1 cytokine or Th1 cells may participate in the pathogenesis of liver damage in chronic HCV infection.     

Reduced expression of the regulatory CD4+ T cell subset is related to Th1/Th2 balance and disease severity in rheumatoid arthritis

OBJECTIVE: To elucidate the involvement of the regulatory CD4+ T cells that produce high levels of interleukin-10 (IL-10) and low levels of IL-4 and IL-2 in the pathogenesis of rheumatoid arthritis (RA), we investigated whether the frequency of this type of CD4+ T cell subset in peripheral blood lymphocytes (PBL) or synovial lymphocyte infiltrates of patients with RA correlated with disease severity and histologic features in rheumatoid synovium. METHODS: PBL and synovial lymphocyte infiltrates were isolated from peripheral blood samples and synovial tissues obtained from 25 patients with RA. Control specimens were obtained from 18 patients with osteoarthritis (OA) and 10 patients with traumatic injuries of the knee joint. CD4+ T cell subsets were categorized as Th1 (production of interferon-gamma [IFNgamma], but not IL-4), Th2 (production of IL-4, but not IFNgamma), or CD4+ T cell subsets producing IL-10, IL-2, or IL-4. The percentages of these T helper subsets among PBL and among synovial infiltrating lymphocytes were determined by an intracellular staining assay with flow cytometric analysis. RESULTS: The level of expression of CD4+ T cells producing IL-10 but not IL-2 and IL-4 in the peripheral blood and synovial tissue was significantly lower in RA patients than in OA patients and trauma patients. In RA patients, the frequency of this type of CD4+ T cell subset among synovial infiltrating CD4+ T cells was inversely correlated with the frequency of Th1 cells and the Th1/Th2 balance in synovial lymphocytes, serum C-reactive protein value, disease activity score, and the degree of synovial lining hyperplasia and lymphocyte infiltration in rheumatoid synovium. There was a reciprocal relationship between the frequency of Thl cells and CD4+ T cells producing IL-10 but not IL-2 and IL-4 in the peripheral blood of RA patients. CONCLUSION: In RA, reduced expression of the CD4+ T cell subset producing IL-10 but not IL-2 and IL-4 may be responsible for the dominance of Th1 over Th2 cells at sites of inflamed synovium and in the peripheral blood. Decreases in this type of CD4+ T cell subset may induce the down-regulation of T cell tolerance and exacerbate the inflammatory process in RA.     

姜黄素对NOD糖尿病小鼠Th1细胞和CD4^+Treg细胞的作用

目的研究姜黄素对Th1细胞和CD4^+Treg细胞的作用,探讨姜黄素对非肥胖糖尿病(NOD)小鼠的治疗作用。方法将10周龄雌性NOD小鼠分为对照组和姜黄素组,分别用溶剂或姜黄素灌胃。第14周时测血糖。第20周时,流式细胞术分析外周血Th1细胞和CD4^+Treg细胞,分析胸腺CD4^+ CD8^-T细胞、CD8+CD4^-T细胞及CD4^+Treg细胞;分选脾CD4^+Treg细胞用于检测其抑制功能。结果姜黄素组血糖正常比例高于对照组。姜黄素组Th1细胞比例显著低于对照组(P<0.05),CD4^+Treg细胞比例与对照组差异无统计学意义(P>0.05),CD4^+Treg/Th1显著高于对照组(P<0.05)。姜黄素组胸腺CD4^+CD8^-T细胞、CD8^+CD4^-T细胞及CD4^+Treg细胞比例与对照组差异无统计学意义(P>0.05),胸腺总细胞数显著高于对照组(P<0.05)。姜黄素组CD4^+Treg细胞的抑制功能明显高于对照组(P<0.05)。结论姜黄素降低NOD小鼠外周Th1细胞、增加CD4^+Treg细胞抑制功能、缓解胸腺退化,延缓了NOD小鼠的高血糖症状。     

T-regulatory cell/T helper 17 cell imbalance functions as prognostic biomarker of oral squamous cell carcinoma – CONSORT

Background:The role of T cells in the pathogenesis of oral squamous cell carcinoma (OSCC) was clarified by examining the levels of differentiated CD4⁺CD25⁺FOXP3⁺ T-regulatory cells (Treg cells) and CD4+IL17+ T helper 17 (Th17) cells in OSCC patients.Methods:Flow cytometry was conducted to measure the proportions of Treg/Th17 cells in different sample groups to identify a proper maker indicative of the progression and prognosis of OSCC.Results:The results showed that a higher Treg/Th17 ratio led to poorer prognosis. Also, the proportions of both Treg cells and Th17 cells were significantly elevated in OSCC patients compared with those in the control groups, suggesting a correlation between Treg/Th17 imbalance and the prognosis of OSCC. Furthermore, the ratios of Treg/Th17 cells in OSCC patients differed at different time points of cancer progression. For example, stage III-IV patients showed the most evident increase in the Treg/Th17 ratio, while the Treg/Th17 ratio in control subjects was the lowest.Conclusions:Therefore, a higher ratio of Treg/Th17 indicated the progression of OSCC and a larger tumor size. Therefore, Treg and Th17 imbalance was implicated in OSCC progression.     

Mechanism of T cell hyporesponsiveness to HBcAg is associated with regulatory T cells in chronic hepatitis B

AIM: To study the mechanisms of hyporesponsiveness of HBV-specific CD4 T cells by testing TH1 and TH2 commitment and regulatory T cells. METHODS: Nine patients with chronic hepatitis B were enrolled. Peripheral blood mononuclear cells were stimulated with HBcAg or HBsAg to evaluate their potential to commit to TH1 and TH2 differentiation. HBcAg-specific activity of regulatory T cells was evaluated by staining with antibodies to CD4, CD25, CTLA-4 and interleukin-10. The role of regulatory T cells was further assessed by treatment with anti-interleukin-10 antibody and depletion of CD4 CD25 cells. RESULTS: Level of mRNAs for T-bet, IL-12R p2 and IL-4 was significantly lower in the patients than in healthy subjects with HBcAg stimulation. Although populations of CD4 CD25highCTLA-4 T cells were not different between the patients and healthy subjects, IL-10 secreting cells were found in CD4 cells and CD4 CD25 cells in the patients in response to HBcAg, and they were not found in cells which were stimulated with HBsAg. Addition of anti-IL-10 antibody recovered the amount of HBcAg-specific TH1 antibody compared with control antibody (P < 0.01, 0.34%±0.12% vs 0.15%±0.04%). Deletion of CD4 CD25 T cells increased the amount of HBcAg-specific TH1 antibody when compared with lymphocytes reconstituted using regulatory T cells (P < 0.01, 0.03%±0.02% vs 0.18%±0.05%). CONCLUSION: The results indicate that the mechanism of T cell hype-responsiveness to HBcAg includes activation of HBcAg-induced regulatory T cells in contrast to an increase in TH2-committed cells in response to HBsAg.