转录因子T-bet在Th1/Th2分化中的作用及意义

T-bet是属于T-box家族的新型转录因子,选择性地表达于Th1细胞,作为Th1特异的转录因子能诱导IFN-γ的产生,在Th1细胞的分化中起着决定性的作用。T-bet是IFN-γ基因强有力的转录激活剂,又能诱导并保持IL-12Rβ2的表达,且能将分化中的效应性Th2和已完全分化的Th2逆转为Th1,伴随产生大量的IFN-γ,并抑制IL-4、IL-5、IL-13等Th2型细胞因子的产生。T-bet在T细胞活化过程中能被IFN-γ诱导而上调;通过STAT1依赖的途径,TGF-β能通过抑制T-bet而抑制Th1分化。T-bet与Th1/Th2相关疾病的联系正日益受到关注。     

MiR-143在Lewis肺癌小鼠外周血中的表达及其对Th1/Th2平衡的影响

目的 利用小鼠Lewis肺癌移植肿瘤模型探究肺癌疾病进展过程中外周血微小RNA的表达及其对Th1/Th2平衡的调控作用。方法 利用Real-time PCR实验检测血浆中miR-143、miR-217、miR-195和miR-615的表达水平;用酶联免疫吸附实验（ELISA）检测小鼠外周血血清IFN-γ、IL-2、IL-4和IL-10表达水平,用流式细胞术检测小鼠脾脏CD4+IFN-γ+Th1细胞及CD4+IL-4+Th2细胞比例,测量并统计荷瘤小鼠皮下接种部位肿瘤体积及肺部转移性结节;分离小鼠外周血CD4 T细胞,检测转录因子T-bet和GATA3的表达。结果 与Normal组相比,LCC组小鼠外周血血清miR-143表达水平显著上升,miR-217、miR-195和miR-615表达水平没有明显变化。与Normal组相比,LCC组小鼠外周血Th1相关细胞因子表达水平及脾脏Th1细胞比例下降,而Th2相关细胞因子表达水平上升,且Th1/Th2比率显著下降。而Anti-miR-143组Th1相关细胞因子及Th1细胞比例升高,且Th2相关细胞因子比例下降。与LCC组相比,Anti-miR...     

白藜芦醇调节Th细胞失衡与T-bet、GATA-3途径作用机制的探讨

目的 探讨白藜芦醇对Jurkat T细胞T-bet/GATA-3表达的影响及其与调节Th1/Th2细胞分化作用机制之间的关系.方法 不同浓度的白藜芦醇刺激Jurkat T细胞48 h后,用ELISA法检测Th1/Th2细胞因子表达谱及用RT-PCR检测T-bet和GATA-3 mRNA表达的变化.结果 白藜芦醇抑制T-bet mRNA和GATA-3 mRNA的表达（F=6308,P〈0.05 F=3262,P〈0.05）.对Jurkat T细胞分泌细胞因子IFN-γ、IL-10的表达均起抑制作用,并具有浓度效应（F=2604,P〈0.05 F=2465,P〈0.05）.结论 白藜芦醇抑制Jurct T细胞由Th0向Th1细胞分化可能是通过降低转录因子T-bet表达水平进行调节,对Th2细胞的抑制作用则可能通过抑制转录因子GATA-3途径进行负调节,并具有浓度效应.     

牛蒡子苷元抑制TLR4/TRAF6/NF-κB信号通路对哮喘模型大鼠气道重塑和Th1/Th2免疫平衡的影响北大核心CSCD

目的研究牛蒡子苷元抑制Toll样受体4(TLR4)/肿瘤坏死因子受体相关因子-6(TRAF6)/核转录因子-κB(NF-κB)信号通路对哮喘模型大鼠气道重塑和Th1/Th2免疫平衡的影响。方法构建急性哮喘大鼠模型,随机分为5组(每组12只模型大鼠):模型组、牛蒡子苷元低、高剂量组、脂多糖(LPS,TLR4激活剂)组、牛蒡子苷元+LPS组,以12只Wistar正常大鼠作为对照组。分组处理后,观察各组大鼠哮喘症状并做评分、分类计数各组大鼠支气管肺泡灌洗液(BALF)中炎症细胞、检测各组大鼠肺组织病理变化、血清IgE水平、BALF和血清中白细胞介素(IL)-4、干扰素γ(IFN-γ)水平、IFN-γ/IL-4及肺组织TLR4/TRAF6/NF-κB通路相关蛋白表达。结果与对照组相比,模型组大鼠肺组织产生严重病理损伤,哮喘症状评分、WAt/Pbm、WAm/Pbm、BALF中巨噬细胞及淋巴细胞计数、血清IgE水平、BALF与血清中IL-4水平、肺组织TLR4、TRAF6表达及p-NF-κB p65/NF-κB p65升高(P<0.05),BALF与血清中Th1型细胞因子IFN-γ水平、IFN-γ/IL-4降低(P<0.05);与模型组相比,各牛蒡子苷元处理组中模型鼠的上述改变均被扭转,且高剂量牛蒡子苷元作用更强;LPS组大鼠各指标变化与牛蒡子苷元处理组相反,另外LPS可逆转高剂量牛蒡子苷元对大鼠各指标的作用。结论牛蒡子苷元可通过抑制TLR4/TRAF6/NF-κB信号激活而阻止哮喘大鼠气道炎症,促使Th1/Th2免疫平衡向Th1转移,改善大鼠气道重塑和肺损伤。     

类风湿关节炎患者血清和关节液增加体外培养健康人PBMC中Th1/Tc1比例的研究

探讨类风湿关节炎(rheumatoid arthritis,RA)患者血清和关节液干预对体外培养的健康人PBMC分泌IFN-γ的影响及临床意义。本研究采用Bio-plex法检测RA患者及健康人血浆中37种炎性细胞因子及可溶性受体的含量;分别用RA患者血清(体积分数为20%)和关节液(体积分数10%)处理体外分离培养的健康人PBMC 5d,流式细胞术分析CD3+CD8-IFN-γ+(Th1)和CD3+CD8+IFN-γ+(Tc1)的细胞含量变化;real-time PCR技术检测转录因子T-bet mRNA的表达变化。结果发现RA患者血浆中多种炎性细胞因子和可溶性受体含量升高;RA患者血清和关节液处理后的健康人PBMC中Th1和Tc1比例升高;Th1和Tc1相关的转录因子T-bet mRNA表达上调。这说明RA患者血清和关节液可使体外培养的健康人PBMC中Th1和Tc1比例增加,原因可能与RA患者血清及关节液中异常存在的炎性因子和其他可溶性介质有关。     

T细胞疫苗对CIA的免疫调节作用

目的探讨T细胞疫苗(TCV)对小鼠胶原诱导性关节炎(CIA)的免疫调节作用。方法通过TCV干预CIA模型,观察小鼠发病情况,运用免疫组化技术观察局部关节病变程度;FACs分析T细胞亚群变化;ELISA检测细胞培养上清中TGF-β、IFN-γ、IL-17水平;定量PCR方法检测Treg、Th1、Th17细胞相关转录因子表达水平。结果经过TCV干预,小鼠自身反应性T细胞的增殖能力被显著抑制,CII抗体水平也显著下降;相较CIA组,TCV组Treg细胞百分比上升,Th1、Th17细胞百分比明显降低;各细胞分泌细胞因子的能力也发生相应的变化,其中TGF-β上升,IFN-γ、IL-17降低;Treg细胞转录因子Foxp3基因水平提高,Th1、Th17细胞转录因子T-bet、RORα及RORγt的mRNA水平降低。结论 TCV干预可通过抑制CIA小鼠自身反应性T细胞增殖,上调小鼠体内Treg细胞并抑制Th1、Th17细胞的表达而发挥免疫调节作用,有效降低小鼠发病严重度和关节炎症。     

结核分枝杆菌耐热抗原激活的人γδT细胞Th2极性分化特征以及T-bet/GATA-3对分化的调控作用

目的研究结核杆菌耐热抗原(MTB-HAg)激活的人外周血γδT细胞Th2极性分化特点和表达于T细胞中的T盒(T-bet)和GATA结合蛋白3(GATA-3)转录因子的调控作用。方法用MTB-HAg刺激人外周血单个核细胞(PBMC)获得MTB-HAg激活的T细胞(MTBAT),分别在中性条件和Th2极化条件培养后,再经10 ng/m L佛波醇酯(PMA)、500 ng/m L离子霉素(ionomycin)和2.5μmol/L莫能菌素(monensin)刺激6 h,四色荧光抗体染色流式细胞术检测γδT细胞和αβT细胞内细胞因子γ干扰素(IFN-γ)和白细胞介素4(IL-4)的表达。流式细胞术分选28 d MTBAT中的γδT细胞和CD4+T细胞,反转录PCR(RT-PCR)检测T-bet和GATA-3 mRNA的表达。结果在中性条件和Th2极化条件下培养的MTBAT中,γδT细胞一直优势表达IFN-γ,而Th2极化条件下,随培养时间增加IFN-γ+αβT细胞百分率显著下降。与中性条件培养比较,Th2极化条件下MTBAT培养28 d,Th0型(IFN-γ+IL-4+)γδT细胞显著增加;而Th2型(IFN-γ-IL-4+)αβT细胞显著增加。RT-PCR结果显示,Th2极化的γδT细胞仍高表达T-bet mRNA,而在CD4+T细胞T-bet mRNA表达被显著下调;同时,GATA-3 mRNA在γδT细胞和CD4+T细胞的表达均显著上调。结论 Th2极化条件下,大部分γδT细胞仍为Th1型,仅部分极性分化为Th0型细胞;Th2极性分化的γδT细胞转录因子T-bet和GATA-3未表现出交互调节功能。     

Galectin-9对活化的CD4+T细胞免疫调节作用的机制研究

目的 研究Galectin-9对活化的CD4+T细胞的免疫调节作用,并进一步探讨其作用机制.方法 获取野生型C57BL/6小鼠淋巴细胞,利用MACS分选CD4+ na(i)ve T细胞,给予anti-CD3 (2.5 μg/ml)抗体、anti-CD28(5μg/ml)抗体和IL-2(100 ng/ml)刺激,培养3d.将活化的CD4+T细胞分为3组:正常对照组、Galectin-9组和Galectin-9+α-乳糖组.利用CFSE检测T细胞增殖情况,并动态观察细胞形态变化;检测CD4+ CD69+T细胞、Th1、Th2和Th17细胞的比例;并利用ELISA检测淋巴细胞分泌IFN-γ 、IL-4、IL-10、IL-12、IL-17A和TGF-β1等细胞因子的水平;利用Western blot检测T-bet、GATA-3和ROR-yt等T细胞分化调控蛋白的变化.结果 与正常对照组和Galectin-9+α-乳糖组相比,Galectin-9组于2h时开始出现细胞形态改变,同时CD4+ CD69+T细胞、Th1和Th17细胞表达减少(P＜0.05),但Th2细胞无明显变化;培养上清液中IFN-γ、IL-12、IL-17A和TGF-β1的表达水平降低(P＜0.05),而IL-4和IL-10等Th2型细胞因子水平无明显变化;同时T-bet和ROR-γt的表达水平减少(P＜0.05).结论 Galectin-9抑制活化CD4+T细胞的Th1和Th17型免疫应答,而对Th2型免疫应答无影响,免疫调节的机制可能与其在转录水平影响Th1和Th17特定转录因子的表达有关.     

以T-bet/GATA-3的比率作为评价哮喘患者Th1/Th2失衡指标的探讨

目的：探讨哮喘患者PBMCs中转录因子T-beL／GATA-3的比率与Th1／Th2细胞失衡的关系。方法：采用RT-PCR法测定30例发作期哮喘患者(哮喘组)及20例慢性阻塞性肺病患者(COPD组)及20例正常人(对照组)PBMCs中T-bet和GATA-3的活性，ELISA法测定血浆IL-4、IL-5、IL-13和IFN-γ的表达水平。结果：哮喘组PBMCs中T-bet／GATA-3的比率较COPD组和对照组明显降低，Th2细胞因子IL-4、IL-5、IL-13的水平增高，与T-bet／GATA-3的比率成负相关，而Th1细胞因子IFN-γ的水平降低，与T-bet／GATA-3的比率成正相关。结论：哮喘患者T-bet／GATA-3的比率降低，可以作为评价Th1／Th2细胞失衡的精确指标。     

Tim-3/galectin-9调控脂多糖诱导的人淋巴细胞中Th1/Th2细胞平衡

目的 探讨T细胞免疫球蛋白黏蛋白分子-3(Tim-3)/半乳糖凝集素-9(galectin-9)在脂多糖(LPS)诱导淋巴细胞炎性过程中对辅助性T细胞(Th)Th1/Th2细胞失衡中的作用。方法 收集脓毒症患者的外周血。分离外周血单个核细胞(PBMC),RT-qPCR检测Tim-3、galectin-9、T-bet和GATA3的表达；ELISA检测血清sTim-3、galectin-9、IFN-γ和IL-4的水平；分离的外周血单个核细胞体外培养，LPS刺激建立体外炎性模型；ELISA法检测细胞培养上清中Th1和Th2细胞相关细胞因子的水平；Western blot检测JAK2/STAT3相关因子的表达；Pearson相关系数分析Tim-3、galectin-9、IFN-γ和IL-4的相关性。结果 脓毒症患者外周血中Tim-3和galectin-9的mRNA表达增多，sTim-3、galectin-9、IL-4和GATA-3升高，IFN-γ和T-bet水平下降，磷酸化JAK2和STAT3升高(P<0.05)。阻断Tim-3表达后，IFN-γ水平升高，而IL-4的水平明显下降，磷酸化...     

Th1、Th2、Th17型细胞因子与糖尿病肾病相关性的研究

糖尿病肾病(diabetic Nephropathy,DN)是导致终末期肾脏病(end-stage renal disease,ESRD)的最主要病因,因此,早期诊断和治疗是糖尿病肾病的诊治要点。但目前糖尿病肾病的确切发病机制尚未完全明确,糖尿病肾病的发生发展与血流动力学改变和代谢的紊乱、氧化应激和炎症等多种因素有关,而细胞因子在2型糖尿病及其相关肾脏并发症的病因、发病机制中也起着重要的作用,各种细胞因子的识别将为糖尿病肾病的诊治提供新的潜在治疗靶点。文章就Th1、Th2、Th17型细胞因子与DN相关性的研究作了综述。     

Th1, Th2, and Th17 cytokines in systemic lupus erythematosus

Abstract

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the breakdown of immune tolerance leading to excessive inflammation and tissue damage. Imbalance in the levels of cytokines represents one of the multifactorial causes of SLE pathogenesis and it contributes to disease severity. Deregulated levels of T helper type 1 (Th1), type 2 (Th2), and type 17 (Th17) cytokines have been associated with autoimmune inflammation. Growing evidence has shown deregulated levels of Th1, Th2, and Th17 cytokines in SLE patients compared to healthy controls associated with disease activity and severity. In this review, we describe and discuss the levels of Th1, Th2, and Th17 cytokines in SLE patients, and clinical trials involving Th1, Th2, and Th17 cytokines in SLE patients. In particular, with the exception of IL-2, IL-4, and TGF-β1, the levels of Th1, Th2, and Th17 cytokines are increased in SLE patients associated with disease severity. Current phase II or III studies involve therapeutic antibodies targeting IFN-α and type I IFN receptor, while low-dose IL-2 therapy is assessed in phase II clinical trials.     

Distribution of Th1, Th2, and Th0 and the Th1/Th2 cell ratios in human peripheral and endometrial T cells

PROBLEM: To examine whether normal pregnancy involves type 2 T-helper (Th2) immune condition or not. METHOD OF STUDY: We measured the percentage of Th0, Th1, and Th2 and the Th1/Th2 cell ratios of human peripheral blood and endometrial T cells using flow cytometry, which can analyze both the surface marker CD3, and intracellular cytokines, interleukin 4 (IL-4) and interferon gamma (IFNgamma). RESULTS: No significant differences were found in the percentages of Th1, Th2, and Th0 and the Th1/Th2 cell ratios in the peripheral blood T cells of nonpregnant women and women in early pregnancy. On the other hand, the percentage of Th1 cells was highest during the proliferative phase of the endometrium, followed by the secretory phase and early pregnancy decidua. The percentage of Th2 cells was highest in early pregnancy decidua and lowest during the proliferative phase of the endometrium. The Th1/Th2 ratio was 147.48+/-96.68 during the proliferative phase of the endometrium, 37.74+/-21.33 during the secretory phase, and 1.31+/-0.48 in the early pregnancy decidua. CONCLUSIONS: These data indicate that Th1 cells predominate in the nonpregnant endometrium, especially during the proliferative phase, while Th2 cells predominate in early pregnancy decidua.     

Toll样受体与Th1和Th2型免疫应答

Toll样受体是广泛表达在哺乳动物细胞表面的跨膜信号传导受体,其通过识别多种类型的病原体相关分子模式及一些内源性配体,激活天然免疫系统,同时通过诱导树突状细胞分化成熟,调控获得性免疫反应的建立。大多数TLRs的配体可诱导机体产生Th1型免疫应答,然而在某些条件下也可导致Th2型免疫应答的发生。弄清TLRs参与调节Th0分化的机制,可为今后在感染免疫、自身免疫、超敏反应等方面进行深入研究及相关疾病的治疗提供新的切入点。     

Th17及Th17型细胞因子与炎症性肠病

Th17细胞已被划分为一个不同于Th1、Th2和Treg的新的T细胞亚群,以分泌IL-17为主要特征。Th17在防御胞外细菌感染、介导慢性炎症和自身免疫性疾病的发病机制中发挥重要作用。炎症性肠病属于自身免疫性疾病的一种,免疫调节紊乱是其发病的关键因素。免疫学和基因学的发现表明Th17及Th17效应因子在炎症性肠病发病机理中起重要作用。对Th17的进一步深入研究可以加深我们对相关疾病发病机制的认识并指导临床治疗。     

Quantitative analysis of peripheral blood Th0, Th1, Th2 and the Th1:Th2 cell ratio during normal human pregnancy and preeclampsia.

We calculated the percentage of Th1, Th2, Th0 cells and the Th1:Th2 cell ratio of peripheral blood from normal pregnant subjects and preeclampsia patients using flow cytometry which can analyse both the surface marker, CD4, and intracellular cytokines, interleukin (IL)-4 and interferon (IFN)-gamma. In normal pregnancy, the percentage of Th1 cells was significantly lower in the third trimester, and the ratios of Th1:Th2 were significantly lower in the second and third trimester than in nonpregnant subjects. In contrast, the percentage of Th1 cells and the ratios of Th1:Th2 in preeclampsia were significantly higher than in normal third trimester pregnant subjects. The percentage of Th2 cells in preeclampsia was significantly lower than in third trimester of normal pregnancy. Additionally, peripheral blood mononuclear cells from these subjects and patients were cultured with phytohemagglutinin stimulation, and IL-4 and IFN-gamma concentrations were determined in the supernatant by enzymed linked immunosorbent assays. The percentage of Th1 and Th2, and the ratios of Th1:Th2 were correlated with cytokine (IFN-gamma and IL-4) secretion level. These results demonstrated that Th2 cells were predominant in the second and third trimesters of normal pregnancy, but Th1 cells predominated in preeclamptic patients.     

Th细胞及其分化调节

幼稚CD4^＋T细胞可分化为Th1和Th2细胞，Th1主要产生IL-2、IFN-γ、TNF，增强吞噬细胞介导的抗感染机制，促进细胞免疫，也在器官特异性自身免疫疾病中起作用；Th2细胞主要产生IL-4、IL-5、IL-10、IL-13，促进B细胞增殖、分化和产生抗体，增强B细胞介导的体液免疫应答，在变态反应和机体抗寄生虫免疫中发挥作用。Th细胞分化主要由局部环境中的细胞因子及细胞内关键转录因子调控。转录因子STAT1、STAT4、IRF1和T—bet促使Th1细胞分化；转录因子STAT6、IRF4和GATA-3促使Th2细胞分化。     

CCK-8对KLH免疫小鼠脾细胞Th1/Th2平衡的影响

目的： 探讨八肽胆囊收缩素（CCK-8）对Th1/Th2平衡的调节作用。方法: 给予BALB/c小鼠钥孔戚血蓝蛋白（KLH）免疫同时体内给予不同剂量的CCK-8,酶联免疫吸附试验(ELISA)检测其脾细胞培养上清中Th1型细胞因子γ-干扰素(IFN-γ)、白细胞介素-2（IL-2）和Th2型细胞因子白细胞介素-4（IL-4）、白细胞介素-5（IL-5）水平,逆转录聚合酶链式反应(RT-PCR)法检测脾细胞中IFN-γ、IL-2、IL-4、IL-5 mRNA表达;ELISA法检测血清中Th1型抗KLH抗体IgG2a和Th2型抗KLH抗体IgG1水平。结果: ①KLH免疫使小鼠脾细胞分泌Th1/Th2型细胞因子水平明显增高,mRNA表达增高,KLH免疫同时给予CCK-8可使脾细胞培养上清中IFN-γ、IL-2含量进一步增加和IFN-γ、IL-2mRNA表达增高,而使IL-4、IL-5含量降低,IL-4、IL-5 mRNA表达减低和降低IL-4/IFN-γ比值。②KLH免疫小鼠血清中IgG2a、IgG1发生不同程度增高,CCK-8可使其血清中IgG1水平减低而使IgG2a水平增高。结论: CCK-8可促进KLH免疫小鼠体内Th1反应,使Th2优势反应向Th1方向转变。     

Th1 and Th2 in human IVF pregnancy with allogenic fetus

PROBLEM: Positive obstetric outcome of allogenic in vitro fertilization (IVF) pregnancies makes interesting the subject of additional regulatory mechanisms after oocyte donation. METHOD OF STUDY: Eighty eight women: 23 donation of oocytes (DO), 33 IVF, 32 natural conception (NC) were studied in first trimester of pregnancy. Intracellular production of cytokines and chemokine receptors expression were studied by flow cytometry. RESULTS: Intracellular production of interferon-gamma (IFN-gamma), interleukin (IL)-4, tumor necrosis factor-alpha by CD4 T lymphocytes in DO women was higher than in IVF and NC women. Ratio IFN-gamma/IL-4 in DO was lower than in IVF. We found higher expression of chemokine receptor CCR4 but not CXCR3 on CD4 T cells in DO compared with IVF and NC. Ratio CXCR3/CCR4 in DO was lower than in NC. CONCLUSION: Hyperactivation of T helper 1 (Th1) and T helper 2 (Th2) by allogenic fetus is specific for DO pregnancy. Preferable activation of Th2 and relative suppression of Th1 chemokine expression reflect additional regulatory counteractive mechanism(s).     

自身免疫病的Th1/Th2细胞因子平衡治疗

在很多自身免疫性疾病中存在着细胞因子不平衡的现象，导致机体细胞免疫或体液免疫的失调。Ｔｈ１／Ｔｈ２分别介导细胞免疫和体液免疫，可以代表机体复杂的细胞因子网络，从整个评估机体免疫功能。