慢性丙型肝炎患者血清细胞因子水平的变化及意义

目的:检测慢性丙型肝炎患者血清细胞因子IL-2、IFN-г、IL-5、IL-6、IL-12P70和P40水平,探讨其与患者血清ALT水平、HVC RNA载量、HCV基因型及干扰素疗效的关系。方法:检测30例健康对照者和30例慢性丙型肝炎患者干扰素治疗前后血清IL-2、IFN-г、IL-5、IL-6、IL-12P70和P40的含量,比较干扰素治疗应答组和无应答组之间细胞因子水平的差异及上述细胞因子水平与血清ALT水平、HCV基因型、HCVRNA载量等的关系。血清细胞因子检测应用ELISA法,HCV基因分型应用直接测序法,HCVRNA载量采用荧光定量PCR法。结果:与健康对照组相比,慢性丙型肝炎患者血清IL-2含量明显降低,IL-5和IL-12P40明显生高;血清IL-6含量与血清ALT水平呈正相关,与RNA载量呈负相关;HCV基因型1型患者血清IL-6含量明显高于2型,其他基因型和亚型之间细胞因子水平均无显著性差异;干扰素治疗的持续应答率为46.7%,应答组和无应答组治疗前血清细胞因子水平均无显著性差异,但应答组治疗结束时IFN-γ含量较治疗前明显升高。结论:血清Th1/Th2细胞因子水平失衡与丙型肝炎的慢性化和肝脏炎症活动相关;干扰素治疗前血清Th1/Th2细胞因子水平与干扰素治疗效果无关,不能对疗效进行预测,干扰素诱导的Th1细胞优势反应与持续应答有关。     

慢性乙型肝炎患者核苷(酸)类似物治疗前后外周血中Treg和Th17细胞及其相关细胞因子水平的变化

目的 观察慢性乙型肝炎(CHB)患者应用核苷(酸)类似物(NA)抗病毒治疗前后外周血中Treg、Th17细胞及其相关细胞因子水平的变化.方法 采用流式细胞术,检测44例NA治疗12周的CHB患者外周血Treg(CD4+ CD25high CD127low)和Th17(CD3+ CD8-IL-17+)细胞频率.ELISA检测血清IL-10、TGF-β1、IL-17及IL-23水平.各组间采用Mann-Whitney U检验,治疗前后采用Wilcoxon配对T检验.结果 完全应答的14例患者的Treg细胞频率、IL-10、TGF-β1和IL-23水平及Treg/Th17比率较治疗前明显下降(Z=-2.691,-2.417,-2.237,-2.291,-2.291,P均＜0.05),Th17细胞频率及IL-17水平略有增加.部分应答的11例患者的Treg细胞频率、IL-10、TGF-β1和IL-23水平较治疗前明显下降(Z=-1.988,-2.934,-2.756,-2.803,P均＜0.05),Th17细胞频率及IL-17水平略有增加,Treg/Th 17比率略有下降.无应答的15例患者的Treg和Th17细胞频率、IL-10、TGF-β1、IL-23、IL-17水平及Treg/Th17比率较治疗无明显变化.HBV DNA下降≥2 log拷贝/ml及ALT恢复正常时,Treg细胞频率与细胞因子IL-10、TGF-β1和IL-23水平较治疗前明显下降(P均＜0.05).Treg细胞频率与HBV DNA及ALT水平三者之间具有一定的相关性.结论 NA抗病毒治疗前后,CHB患者Treg和Th17细胞频率及IL-10、TGF-β1、IL-23、IL-17水平均发生变化;治疗后获得满意应答的CHB患者Treg细胞频率、IL-10、TGF-β1、IL-23水平及Treg/Th17比率明显下降,Th17细胞及IL-17水平继续上升;且以上变化与病毒学应答及生化学应答有关.     

慢性丙肝外周血Th17/Th1水平与干扰素联合利巴韦林标准治疗快速应答的关系

目的 探讨Th17、Th1细胞在慢性丙型肝炎病毒感染患者外周血的表达情况及其与干扰素联合利巴韦林标准治疗快速应答的关系.方法 收集31例慢性丙型肝炎(Chronic hepatitis C,CHC)接受干扰素联合利巴韦林标准剂量治疗患者的人口学资料(包括年龄、性别、体重指数等)、肝功能及治疗基线时外周血标本,并以11例健康志愿者为对照组.采用前瞻性实验研究方法,观察CHC治疗4周时获得快速病毒学应答情况,采用流式细胞术检测治疗基线时外周血单个核细胞中CD3+、CD8 -、IL-17+、Th17细胞和CD3+、CD8-、IFN-γ+、Th1细胞的比例.结果 CHC患者较健康对照者外周血中Th1水平下降,Th17水平上升;标准抗病毒治疗无快速应答组较健康对照组基线时Th1细胞水平明显下降,Th17细胞水平明显升高(P＜0.05).结论 干扰素联合利巴韦林标准治疗CHC无快速应答者外周血Th1/Th17亚群失衡,Th1水平下调,Th17水平上调;治疗基线时高Th17水平及低Th1水平可能不利于产生快速病毒学应答.     

Th1/Th2漂移与HCV感染慢性化

Thl和Th2细胞分别介导机体的细胞免疫和体液免疫,它们来自一个共同的前体细胞Th0,在不同的细胞因子、抗原等因素的影响下,可发生Thl与Th2的转换。HCV感染时,Th1应答与丙型肝炎的好转有关,Th2应答与丙型肝炎慢性化有关。因此,Thl与Th2之间的漂移对HCV感染的转归有重要影响,是HCV感染慢性化的重要机制。     

米非司酮通过增强母-胎界面Th1型偏移导致流产

目的：探讨米非司酮对母胎界面Th1/Th2型细胞因子动态平衡的影响.方法：将63例早孕期妇女随机分为2组,一组一次服用米非司酮200 mg,另一组为对照组,收集其蜕膜组织.应用免疫组化法,评价Th1型细胞因子（IL-2、IFN-γ）、Th2型细胞因子（TGF-β2、IL-4）的表达.结果：正常妊娠时,在母-胎界面Th2型细胞因子（IL-4）以及TGF-β2的表达较高;Th1型细胞因子（IL-2、IFN-γ）的表达较低,尤其是IL-2.服用米非司酮后,蜕膜Th1型细胞因子（IL-2、IFN-γ）表达显著升高;而母-胎界面Th2型细胞因子（IL-4）以及TGF-β2的表达无明显变化.结论：米非司酮打破了正常妊娠时母胎界面Th2型免疫优势;显著升高Th1型细胞因子（IL-2、IFN-γ）表达,形成了Th1型免疫偏离,导致流产的发生.     

Th3、Th17及相关细胞因子在自身免疫性甲状腺疾病发病中的作用

目的:研究Th17、Th3细胞及相关细胞因子IL-17、TGF-pI在自身免疫性甲状腺疾病(AITD)发病机制中的作用及意义.方法:收集49例初诊的自身免疫性甲状腺疾病患者,分为Graves’病(GD)(n=30)组和桥本甲状腺炎(HT)(n=19)组,另选年龄、性别匹配的18个正常人作为健康对照.Graves’病组采用甲巯咪唑治疗,桥本甲状腺炎组采用甲状腺素钠治疗,均随访至甲状腺功能正常.采用流式细胞仪技术检测外周血中Th17和Th3细胞比例.采用ELISA方法检测血浆IL-17、TGF-β1水平.结果:GD和HT初诊组患者外周血Th17细胞较正常对照组明显升高(P＜0.05).Th3细胞在HT初诊组中明显升高,而其在GD初诊组中明显下降(P＜0.05).血浆IL-17水平在GD和HT初诊组、缓解组均较正常对照组明显升高(P＜0.05).血浆TGF-β1水平在HT初诊组和缓解组均明显升高(P＜0.05).结论:Th17、Th3细胞和相关细胞因子IL-17、TGF-β1可能参与了自身免疫性甲状腺疾病的发病,TGF-β1可能在HT的发病中起了一个促炎因子的作用.     

全氟化合物与儿童哮喘及Th1/Th2免疫应答平衡相关性研究

目的 采用病例对照探讨全氟化合物(PFAAS)暴露与儿童哮喘及Th1型细胞因子白细胞介素(IL)-2,干扰素(IFN)-γ和Th2型细胞因子(IL-4,IL-5)分泌水平的关系.方法 选择231名台北医院就诊的哮喘儿童作为病例组,来自社区的225名自然儿童作为对照组.采用双抗体酶联免疫吸附实验(ELISA)试剂盒检测儿童血清中细胞因子IL-2、IFN-γ、IL-4和IL-10的分泌水平;高效液相色谱仪分析血清中全氟辛烷磺酸(PFOS)和全氟辛酸(PFOA)水平.结果 哮喘儿童机体PFOS(33.9μg/L比28.9 μg/L)和PFOA(1.2μg/L比0.5 μg/L)暴露负荷显著的高于对照组儿童,且随着机体PFAAs的增高,儿童患有哮喘的风险呈增高趋势.对哮喘儿童而言,血清PFAAs水平与Th1型细胞因子(IL-2,IFN-γ)存在显著的负相关,而与Th2型细胞因子(IL-4,IL-5)呈正相关关系.结论 PFOS暴露可诱导机体免疫应答平衡紊乱,并向Th2型免疫应答极化.     

不明原因习惯性流产患者主动免疫治疗前后Th1/Th2型细胞因子水平的变化

为探讨主动免疫治疗对不明原因习惯性流产 (UHA )患者Th1/Th2型细胞因子水平的影响。采用酶联免疫吸附法检测15例正常非妊娠妇女、 35例UHA患者淋巴细胞主动免疫治疗前后经滋养细胞抗原刺激的外周血单个核细胞 (PBMC )培养上清液中IL 2、IFN γ、IL 4、IL 10的水平。结果发现 :(1)在最佳诱导时间下 ,UHA组治疗前PBMC产生IL 2、IFN γ的水平明显高于正常对照组 (P <0 0 5 ) ,IL 4、IL 10水平明显低于正常对照组 (P <0 0 5 )。UHA组治疗后PBMC产生IL 2、IFN γ的水平较治疗前明显降低 (P <0 0 5 ) ,IL 4、IL 10水平较治疗前明显升高 (P <0 0 5 )。UHA组治疗后PBMC产生各细胞因子的水平与正常对照组比较 ,差异均无显著性 (P >0 0 5 ) ;(2 )UHA组 35例患者主动免疫治疗后半年内 2 8例妊娠 ,其中 9例又出现自然流产。 9例自然流产者治疗后IL 2、IFN γ水平未明显下降 ,IL 10水平未明显上升。 19例妊娠成功者治疗后IL 2、IFN γ水平较治疗前明显下降 (P <0 0 5 ) ,IL 4、IL 10水平明显上升 (P <0 0 5 )。以上结果表明UHA患者对滋养细胞抗原产生以Th1型反应为主的免疫应答 ,产生大量Th1型细胞因子 ,主动免疫治疗有助于上调Th2型细胞因子及下调Th1型细胞因子 ,利于UHA患者妊娠成功。     

自身免疫性溶血性贫血患者外周血Th1／Th2、Th3／Tr1细胞的检测及意义

目的探讨自身免疫性溶血性贫血（AIHA）患者外周血Th1／Th2、Th3／Tr1细胞状态,分析它们在AIHA发病机制中的作用。方法收集AIHA患者及健康者外周抗凝静脉血,分离纯化淋巴细胞。运用FITC-CD3单抗,Cy5-CD4单抗,PE—CRTH2单抗,以CD3／CD4设门作三色流式细胞术检测Th1／Th2细胞,ELISA法检测血清中Th3细胞相关的细胞因子TGF-β1的含量和Tr1细胞的相关因子IL-10的含量。结果与正常对照相比．AIHA患者外周血CD3^＋CD4^＋CRTH2-T细胞（Th1）百分率、CD3^＋CD4^＋CRTH2^＋T细胞（Th2）百分率均下降（P〈0．05）,而CD4^＋CRTH2^-T／CD4^＋CRTH2^＋T比例（Th1／Th2）均明显升高（P〈0．01）,Th3／Tr1细胞分泌的相关细胞因子TGF-β1和IL-10的含量均降低（P〈0．05）。结论AIHA患者外周血存在细胞免疫功能失调,T细胞亚群极化状态发生改变,呈Th1型细胞优势,Th3／Tr1细胞因子含量下降,可能与AIHA的免疫学发病机制有关。     

慢性乙型肝炎患者外周血Th17细胞/调节性T细胞比值变化的意义

目的测定慢性乙型肝炎(CHB)患者外周血CD4+CD25+Foxp3+调节性T细胞(Treg)和CD4+IL-17+T细胞(Th17细胞)频率及其相关细胞因子的变化,并探讨Th17/Treg平衡在CHB发病过程中的作用。方法选取CHB住院患者(CHB组)60例,其中35例轻中度CHB患者(CHB-LM),25例慢性重型肝炎患者(CSHB),同时选取21位健康体检者作为正常对照组(HC)。流式细胞术检测外周血中Th17和Treg的细胞频数,双抗体夹心ELISA检测血清中IL-17、IL-23、IL-10及转化生长因子β1(TGF-β1)的表达水平。结果与HC组相比,CHB患者外周血Th17细胞频率及其相关细胞因子IL-17、IL-23浓度明显增高,差异有统计学意义(P0.05);Treg频率及其相关细胞因子IL-10及TGF-β1浓度明显增高。Th17/Treg比值变化显示,与HC组相比,在CHB组中该比例明显升高,具有显著性差异,且与CHSB组相比,在CHB-LM组中该比值明显降低。结论 Th17细胞/Treg的失衡是造成慢性乙型肝炎病程演变的重要因素,检测Th17细胞/Treg比值变化对疾病发展的早期预测有一定价值。     

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/Th2 cytokine gene expression after mercuric chloride in susceptible and resistant rat strains

Mercuric chloride (HgCl₂) has contrasting effects on different rat strains: susceptible strains, e.g. Brown Norway (BN) develop polyclonal B cell activation, multiple autoantibodies and widespread tissue injury. Lewis (LEW) rats are resistant: no autoimmune response occurs after HgCl₂; instead, there is immunosuppression. We have previously shown, by fully quantitative polymerase chain reaction (PCR), up-regulation of interleukin-4 (IL-4) gene expression in HgCl₂-treated BN rats, implicating Th2 cells in the autoimmune syndrome. Involvement of the reciprocal Th1 subset, producing interferon-γ (IFN-γ), in resistance of LEW rats to HgCl₂ has been suggested. We now report extensive analysis of Th1 and Th2 cytokine gene expression in spleen and lymph nodes of susceptible (BN) and resistant (LEW) rats after HgCl₂. IL-4 and IFN-γ were analyzed by quantitative PCR, other cytokines were assessed using semiquantitative PCR: the relative merits of these two techniques are discussed. We show pronounced up-regulation of IL-4 and more modest up-regulation of IFN-γ in BN rats, but no up-regulation of either in LEW rats. Baseline levels of IFN-γ were higher in LEW rats. Semi-quantitative PCR showed increased expression of IL-2, IL-6 and IL-10 in BN; in LEW rats only IL-10 was increased. There was no marked change in IL-5, IL-13 or transforming growth factor-β (TGF-β) in either strain. These data further support the key role of IL-4 in HgCl₂-induced autoimmunity, and suggest that failure of up-regulation of IL-4, together with higher baseline IFN-γ expression, accounts for resistance of LEW rats to HgCl₂. However, neither IFN-γ nor TGF-β can be implicated in HgCl₂-induced immunosuppression in the LEW rat in vivo: our data suggest a role for IL-10 in this phenomenon.     

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.     

Role of transforming growth factor-βthe preferential induction of T helper cells of type 1 by staphylococcal enterotoxin B

Stimulation of murine CD4⁺ T cells with staphylococcal enterotoxin B (SEB) results in the preferential development of T helper (Th) 1 cells [i.e. high interferon (IFN)-γ and low interleukin (IL)-4, IL-5 and IL-10]; whereas in response to plate-bound anti-CD3 or anti-T cell receptor-αβ, Th1 as well as Th2 cells develop. In the present study, we examined the mechanism which is responsible for the selective Th1 development in the SEB system. The addition of IL-4 resulted in a strong development of Th2 cells showing that SEB stimulation can result inTh2 differentiation. Co-stimulation with anti-CD28 was insufficient in this regard. Lack of Th2 development in the SEB system was in part due to the inhibitory effect of endogenously produced transforming growth factor-β (TGF-β), because anti-TGF-β allowed the development of Th2 cells. Similarly, TGF-β inhibited Th2 development and stimulated Th1 development in the anti-CD3 system. This shift was only partially prevented by also including IL-4 in the cultures. The effects of TGF-β could only partially be explained by stimulation of IFN-γ or inhibition of IL-4 as intermediatory cytokines: (1) TGF-β stimulated Th1 development even in the presence of anti-IL-4 and anti-IFN-γ, and (2) a strong inhibitory effect of anti-TGF-β on Th1 development was still observed when anti-IL-4 and IFN-γ were simultaneously added to the cultures. It is concluded that SEB favors Th1 development by stimulation of TGF-β production. Inhibition of Th2 development by TGF-β is due, in part, to inhibition of IL-4 and stimulation of IFN-γ, and, in part, to a direct effect of TGF-β on the responding T cells.     

Relationship between Th1/Th2 cytokine patterns and the arthritogenic response in collagen-induced arthritis

It is hypothesized that the balance of cytokines produced by Th1/Th2 subsets of T helper cells plays an important role in the development of autoimmune diseases. Murine collagen-induced arthritis (CIA) is an example of an autoimmune disease in which immunization with cartilage-derived type II collagen induces, firstly, a T cell response to type II collagen and, secondly, the manifestation of a destructive inflammatory response in affected joints. We have investigated the role of Th1/Th2 responses in the development of CIA by monitoring levels of interferon (IFN)-γ (a Th1 cytokine) and interleukin (IL)-4 and IL-10 (Th2 cytokines), and IL-1β and tumor necrosis factor (TNF) (pro-inflammatory cytokines) produced by cultured draining lymph node cells (LNC) from collagen-immunized DBA/1 mice during the induction phase of arthritis and throughout the time of clinical manifestation and subsequent remission of the disease. Although a transient increase in IL-10 was detected 3 days after immunization, Th2 cytokine production was found to be almost completely suppressed 6 days after immunization. In contrast, IFN-γ was detected in LNC cultures as early as 6 days after immunization and the addition of type II collagen to the culture medium resulted in an approximately 10-fold increase in IFN-γ production, indicating that a predominantly Th1 response had become established by this time. IFN-γ production by LNC was found to be further increased at the time of clinical manifestation of arthritis and could be up-regulated by co-culture with type II collagen. IL-10 was not detected in LNC cultures at the onset of arthritis and IL-4, although present, was found to be markedly suppressed in LNC cultures containing type II collagen. These findings indicate that Th1 responses are predominant at the time of onset of arthritis and that the activation of collagen-specific Th1 cells may result in suppression of Th2 activity. IFN-γ production declined progressively during the progression and subsequent remission of arthritis whereas levels of IL-10 increased and low, though persistent, levels of IL-4 were detected throughout this period. High levels of IL-1β and TNF-α production were detected at the onset of the disease. The role of Th1 responses in the development of CIA was further emphasized by the observation that immunization of mice with type II collagen in incomplete Freund's adjuvant, which normally fails to induce arthritis, resulted in a predominantly Th2 cytokine profile.     

乳杆菌对原代淋巴细胞中Th1/Th2细胞平衡的影响

目的:分析7种乳杆菌对原代淋巴细胞增殖和细胞因子(CK)分泌的作用,进而探讨其对Th1/Th2细胞平衡的影响。方法:用不同种属、不同浓度的活的/热致死的乳杆菌体外作用于小鼠脾淋巴细胞培养60 h后,采用MTT比色法检测淋巴细胞的增殖效果。用ELISA法检测Th1型细胞因子(IL-12、IFN-γ)、Th2型细胞因子(IL-4、IL-10)和调节型细胞因子(TGF-β)的分泌量。结果:活的/热致死的乳杆菌单独作用,就能促进淋巴细胞体外增殖并表现出剂量依赖关系(P<0.05)。当菌的浓度为107集落形成单位(CFU)/mL(即细菌与细胞的比例为10∶1)时,热致死的发酵乳杆菌和嗜酸乳杆菌的免疫活性近似于活菌。而且,这两株热致死菌还可适当提高淋巴细胞分泌IL-12和IFN-γ,抑制IL-4、IL-10和TGF-β的分泌,使其IFN-γ/IL-4的比值(代表Th1/Th2细胞平衡)均显著高于刀豆蛋白A(ConA)对照组(P<0.05)。结论:乳杆菌可通过提高淋巴细胞的IFN-γ/IL-4分泌率来促进Th1优势状态的Th1/Th2细胞平衡,并具有菌株特异性。     

Mast cells promote Th1 and Th17 responses by modulating dendritic cell maturation and function

Mast cells (MCs) play an important role in the regulation of protective adaptive immune responses against pathogens. However, it is still unclear whether MCs promote such host defense responses via direct effects on T cells or rather by modifying the functions of antigen-presenting cells. To identify the underlying mechanisms of the immunoregulatory capacity of MCs, we investigated the impact of MCs on dendritic cell (DC) maturation and function. We found that murine peritoneal MCs underwent direct crosstalk with immature DCs that induced DC maturation as evidenced by enhanced expression of costimulatory molecules. Furthermore, the MC/DC interaction resulted in the release of the T-cell modulating cytokines IFN-γ, IL-2, IL-6 and TGF-β into coculture supernatants and increased the IL-12p70, IFN-γ, IL-6 and TGF-β secretion of LPS-matured DCs. Such MC-"primed" DCs subsequently induced efficient CD4+ T-cell proliferation. Surprisingly, we observed that MC-primed DCs stimulated CD4+ T cells to release high levels of IFN-γ and IL-17, demonstrating that MCs promote Th1 and Th17 responses. Confirming our in vitro findings, we found that the enhanced disease progression of MC-deficient mice in Leishmania major infection is correlated with impaired induction of both Th1 and Th17 cells.     

1型糖尿病患儿CD4~+T细胞亚群变化初探

目的 探讨CD4~+细胞亚群[Th1、Th2、CD4~+CD25~+Foxp3~+调节性T细胞(Tr)及Th17细胞]在1型糖尿病(TIDM)患儿免疫发病机制中的作用.方法 新诊断TIDM患儿20例,同年龄对照组(Ctrl组)20例.用流式细胞术检测外周血Th1、Th2、Tr及Th17细胞比例.荧光定量PCR(real-time PCR)检测Th1、Th2、Tr、Th17细胞转录因子T-bet、GATA-3、Foxp3、ROR-γt、IFN-、IL-4、IL-10、IL-17A、CTLA-4、GITR等细胞因子和负性调节因子mRNA表达;应用酶联免疫吸附方法(ELISA)检测IFN-γ、IL-4、TGF-β、IL-6血浆水平.结果 (1)与正常对照组相比,TIDM患儿Th1细胞比例明显增高(P<0.01),Th2细胞比例明显降低(P<0.01),Tr和Th17细胞比例与正常对照组相比无明显差别(P>0.05).(2)Th1细胞转录因子及细胞因子T-bet、IFN-γ较正常对照组明显升高(P<0.01);Th2细胞转录因子及细胞因子GATA-3、IL-4明显降低(P<0.01);Tr细胞转录因子Foxp3表达与正常对照组相比差异无统计学意义(P>0.05),Tr细胞相关细胞因子及负性调节因子IL-10、CTLA-4及GITR基因表达明显低于对照组(P<0.01);Th17细胞转录因子ROR-γt及细胞因子IL-17A基因表达与对照组相比无明显差异(P>0.05);(3)TIDM患儿外周血IFN-γ浓度明显增高,IL-4明显降低,TGF-β、IL-6浓度无明显改变(P>0.05).结论 TIDM患儿Th1/Th2失衡,加上Tr细胞抑制功能缺陷,可能导致TIDM严重细胞免疫功能紊乱.