CD4+CD25+调节性T细胞与儿科疾病

古希腊特尔斐阿波罗神庙上镌刻着一句铭言:Gnothi Seauton(英文为know thyself)即"认识自己"."认识自己"已成为免疫学家广为认可的定律:机体免疫系统首先必须识别自身的抗原,产生无反应性,再针对外来抗原产生免疫应答,即"识别自身,排斥异己".     

CD4+CD25+ regulatory T cells in health and disease

1. Over the past 5 years, tremendous progress has been made in understanding the suppressive mechanisms of T regulatory (Treg) cells. The Treg cells, a subpopulation of T cells, have been shown to play an important role in maintaining peripheral tolerance and the prevention of autoimmunity. 2. Various populations of Treg cells have been described, including thymically derived CD4(+)CD25(+) Treg cells. These naturally occurring Treg cells are present in the periphery and are capable of suppressing proliferation and effector T cell responses both in vitro and in vivo. 3. In addition, a second subset of Treg cells, type 1 T regulatoary (Tr1) and Th3 cells, exert their suppressive capacity via cytokines such as interleukin-10 and transforming growth factor-beta and are contact independent. 4. The present review summarizes the characteristics and molecular basis of CD4(+)CD25(+) Treg cells, as well as their therapeutic potential in modulating inflammatory diseases, such as inflammatory bowel disease and rheumatoid arthritis.     

孕妇外周血CD4+CD25+调节性T细胞对子痫前期的预测

目的:探讨孕妇外周血CD4+ CD225+调节性T细胞(CD4 CD25+ Tregs)对子痫前期的预测价值.方法:选取190例于我院产前常规孕期体检及分娩的孕产妇作为研究对象,按孕产妇是否发生子痫前期分为子痫前期组及正常对照组,采用流式细胞仪测定各孕妇20～24周外周血单个核细胞(PBMC)中CD4T细胞及CD4 CD25+ Tregs的比率,并随访至分娩.采用磁珠分选技术分选随访期间确诊子痫前期的患者外周血PBMC中CD4 CD25+ Tregs和CD4CD25T细胞,酶联免疫仪检测CD4 CD25+ Tregs对CD4CD25T细胞增殖的抑制作用,并与正常孕妇组对比.结果:①随访情况.190例孕产妇妊娠期内有15例发展为子痫前期,其余175例为正常分娩,2组在剖宫产率、胎儿心率异常率、新生儿1 min及5 min的Apgar评分有显著性差异(P＜0.05).②CD4 CD25+ Tregs的数量.子痫前期组和正常对照组外周血PBMC中CD4T细胞比率分别为34.21％±6.92％和35.72％±7.45％,2组比较无显著性差异(P＞0.05).子痫前期组外周血PBMC中CD4 CD25+ Tregs占CD4T细胞的比率为6.71％±2.21％,明显低于正常对照组的12.01％±2.98％(P＜0.05).③CD4 CD25+ Tregs的抑制功能.子痫前期组CD4 CD25+ Tregs对CD4 CD25-T细胞增殖的抑制百分率为57.56％±9.47％,正常孕妇组为78.27％±12.43％,2组比较有显著性差异(P＜0.05).④CD4 CD25+ Tregs的预测性.以CD4+ CD25+ Trges数量变化预测子痫前期的ROC曲线下面积为0.913,Tregs的最佳截断点为6.605％,预测子痫前期的敏感度86.7％、特异度82.85％.结论:孕妇妊娠中期外周血CD4 CD25+ Tregs数量减少和(或)抑制功能下降与子痫前期的发生相关,可作为子痫前期的预测性指标.     

The generation and antigen-specificity of CD4+CD25+ regulatory T cells

CD4+CD25+ regulatory T cells are essential components of the immune system. They help to maintain immune tolerance by exerting suppressive effects on cells of the adaptive and innate immune system. In the last few years there has been an abundance of papers addressing the suppressive effects of CD4+CD25+ regulatory T cells and their putative role in various experimental disease models and human diseases. Despite the enormous amounts of data on these cells a number of controversial issues still exists. CD4+CD25+ regulatory T cells were originally described as thymus-derived anergic/suppressive T cells. Recent papers however indicate that these cells might also be generated in the periphery. Due to the thymic development of CD4+CD25+ regulatory T cells it was thought that these cells were specific for self-antigens. Indeed it was shown that CD4+CD25+ regulatory T cells could be positively selected upon high affinity interaction with self-antigens. However, evidence is accumulating that these cells might also interact with non-self antigens. Finally, in the literature there is conflicting evidence regarding the role of soluble factors versus cell-contact in the mechanism of suppression. The aim of this review is to summarize the evidence supporting these opposing viewpoints and to combine them into a general model for the origin, function and antigen-specificity of CD4+CD25+ regulatory T cells.     

Targeting CD4+CD25+FoxP3+ regulatory T-cells for the augmentation of cancer immunotherapy

CD4+CD25+FoxP3+ T-regulatory (Treg) cells are vital to the maintenance of peripheral self tolerance and are implicated in tolerance to foreign antigens. Increasing evidence shows that Treg cells may also play an important role in immune evasion mechanisms employed by cancer. Treg cells are actively recruited and induced by tumors to block innate and adaptive immune priming, effector function and memory response, which can inhibit the efficacy of therapeutic cancer vaccines. As such, modulation of Treg cell function in cancer has been studied using various approaches, with encouraging preclinical and clinical findings. However, controlled and effective modulation of Treg cell function for cancer therapeutics will be contingent on a better understanding of the molecular basis of Treg cell interaction with tumor cells and ensuing immunosuppressive mechanisms.     

雷帕霉素诱导Balb/c小鼠CD4~+ CD25~+ foxp3~+调节性T细胞增殖

目的探讨雷帕霉素对Balb/c小鼠CD4+ CD25+ foxp3+调节性T细胞的作用。方法取8wk龄的SPF级Balb/c小鼠30只,随机分为两组,实验组每只灌胃雷帕霉素0.4mg.d-1,对照组灌胃每天予等体积无菌水,共3wk。无菌条件下肝素抗凝心脏采血,分离脾脏,制备单细胞悬液。采用流式细胞仪检测小鼠外周血和脾细胞CD4+CD25+T细胞,实时定量PCR检测小鼠脾细胞foxp3 mRNA的表达。结果实验组小鼠外周血和脾细胞中CD4+CD25+T细胞的比例分别为(9.95±4.65)%和(24.13±10.06)%,对照组小鼠外周血和脾细胞中CD4+CD25+T细胞的比例分别为(5.01±1.49)%和(8.48±3.19)%,差异均有显著性(P<0.01)。实验组小鼠脾细胞foxp3 mRNA的表达水平明显高于对照组,是对照组的6.029倍,差异有显著性(P<0.01)。结论雷帕霉素能够明显诱导Balb/c小鼠体内CD4+CD25+T细胞的增殖,并能提高foxp3+ mRNA的表达。     

Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine

Natural killer (NK) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. Because bone marrow-derived hematopoietic stem cells (HSCs), lymphoid protenitors, can give rise to NK cells, NK ontogeny has been considered to be exclusively lymphoid. Here, we show that porcine c-kit⁺ bone marrow cells (c-kit⁺ BM cells) develop into NK cells in vitro in the presence of various cytokines [interleukin (IL)-2, IL-7, IL-15, IL-21, stem cell factor (SCF), and fms-like tyrosine kinase-3 ligand (FLT3L)]. Adding hydrocortisone (HDC) and stromal cells greatly increases the frequency of c-kit⁺ BM cells that give rise to CD2⁺CD8⁺ NK cells. Also, intracellular levels of perforin, granzyme B, and NKG2D were determined by RT-PCR and western blotting analysis. It was found that of perforin, granzyme B, and NKG2D levels significantly were increased in cytokine-stimulated c-kit⁺ BM cells than those of controls. And, we compared the ability of the cytotoxicity of CD2⁺CD8⁺ NK cells differentiated by cytokines from c-kit⁺ BM cells against K562 target cells for 28 days. Cytokines-induced NK cells as effector cells were incubated with K562 cells as target in a ratio of 100:1 for 4 h once a week. In results, CD2⁺CD8⁺ NK cells induced by cytokines and stromal cells showed a significantly increased cytotoxicity 21 days later. Whereas, our results indicated that c-kit⁺ BM cells not pretreated with cytokines have lower levels of cytotoxicity. Taken together, this study suggests that cytokines-induced NK cells from porcine c-kit⁺ BM cells may be used as adoptive transfer therapy if the known obstacles to xenografting (e.g. immune and non-immune problems) were overcome in the future.     

雷帕霉素诱导大鼠体内CD4+CD25+FoxP3+调节性T细胞的增殖

目的 探讨雷帕霉素(RPM)对大鼠CD4+CD25+FoxP3调节性T细胞的影响.方法 大鼠20只随机均分为两组:实验组RPM 2 mg·kg-1·d-1灌胃2周;对照组用生理盐水替代.无菌件下下腔静脉采血,并分离脾脏及胸腺,制备单个核细胞悬液.采用流式细胞术检测大鼠外周血、脾脏及胸腺内CD4+CD25+T细胞的占单个核细胞的比例,实时定量-PCR检测脾脏细胞FoxP3 mRNA表达,ELISA检测外周血血清转化生长因子β(TGF-13)和白细胞介素10(IL-10)含量.结果 实验组外周血、脾脏和胸腺中CD4+CD25+T细胞的比例明显高于对照组(P＜0.05).实验组大鼠脾脏细胞FoxP3 mRNA表达为对照组的4.1倍.实验组TGF-β和IL-10显著高于对照组(P＜0.05).结论 RPM能诱导大鼠体内CD4+CD25+FoxP3+调节性T细胞增殖,且增加体内免疫抑制性细胞因子TGF-β和IL-10的分泌.     

Resveratrol induces the suppression of tumor-derived CD4+CD25+ regulatory T cells

CD4+CD25+ regulatory T cells (Treg cells) are negative regulator of the immune system and main obstacles to cancer immunotherapy in tumor-bearing hosts. Resveratrol is a natural product found in grapes with both immunomodulatory and anticancer effects, which can be controlled by Treg cells. Therefore, to determine whether resveratrol performs these actions via Treg cells, we investigated changes in Treg cell population and immunomodulatory cytokines in EG7 tumor-bearing C57BL/6 mice. In the present study, CD4+CD25+ cell population among CD4+ cells was inhibited ex vivo by resveratrol treatment in a dose-dependent manner. FoxP3+ expressing cells among CD4+CD25+ population were significantly reduced after resveratrol treatment ex vivo in intracellular FACS analysis. Single intraperitoneal administration of 4 mg/kg resveratrol suppressed the CD4+CD25+ cell population among CD4+ cells and downregulated secretion of TGF-beta, an immunosuppressive cytokine, measured from the spleens of tumor-bearing mice. Furthermore, resveratrol enhanced IFN-gamma expression in CD8+ T cells both ex vivo and in vivo,leading to immune stimulation. Taken together, these results suggest that resveratrol has a suppressive role on CD4+CD25+ cell population and makes peritumoral microenvironment unfavorable to tumor in tumor-bearing mice. Thus, resveratrol can be considered as possible adjuvant material for vaccination-based cancer therapy.     

造血移植物中CD3~+CD8~(low)和CD3~+CD4~-CD8~(low)细胞亚群的检测及分析

目的检测造血移植物(正常人骨髓、脐血及动员后外周血)中CD3+CD8low和CD3+CD4-CD8low细胞亚群,探讨其促进造血干/祖细胞植入异基因骨髓的功能,以期拓宽脐血移植的应用范围。方法直接三色免疫荧光标记流式细胞术检测。结果CD3+CD8low和CD3+CD4-CD8low细胞亚群占CD3+细胞亚群的比例在骨髓中最高,为(8.61±1.40)%,动员后外周血次之,为(5.11±0.76)%,脐血最低,为(3.31±0.88)%(P<0.01);"初始"T(CD45RA+CD45RO-)细胞亚群占CD8low细胞亚群的比例为脐血(94.26±2.46),骨髓(58.68±7.57),动员后外周血(73.21±3.60),"初始"T占CD8high细胞亚群的比例为脐血(82.63±3.16),骨髓(38.69±3.24),动员后外周血(51.58±4.23),各移植物中"初始"T细胞亚群占CD8low细胞亚群的比例均高于其占CD8high细胞亚群的比例(P<0.01),CD8low细胞亚群中"初始"T与"记忆"T(CD45RA-CD45RO+)细胞亚群的比例均高于二者在CD8high细胞亚群的比例。结论脐血CD8low和CD8lowCD4-细胞占CD3+细胞的比例明显低于骨髓,可能是脐血移植植入延迟的原因之一;"初始T与"记忆"T细胞亚群的比例增高,可能与CD3+CD8low细胞亚群维持和诱导免疫耐受,不引起移植物抗宿主病有关。     

CD4+CD25+Foxp3+调节性T细胞在急性白血病患者外周血的表达

目的 初步探讨CD4 CD25 调节性T细胞在急性白血病(AL)患者外周血中的表达及其临床意义.方法 流式细胞术分别检测43例AL初诊(A组)、经化疗完全缓解(CR,B组)25例及20例健康志愿者(C组)外周血中CD4 CD25 T细胞所占比例;荧光定量RT-PCR分析各组外周血中叉状头/翅膀状螺旋转录因子(Foxp3)mRNA的表达水平,并逐层分析比较.结果 B组CD4 CD25 T细胞比例及Foxp3含量均明显高于A、C组(P＜0.01).A组Foxp3表达明显高于C组(P＜0.01).在AL各亚型之间CD4 CD25 T细胞及Foxp3表达均无显著差异.结论 CD4 CD25 调节性T细胞在初治AL或CR患者外周血中比例增加,可能是AL免疫抑制的一个重要原因.     

CD8+CD103+ regulatory T cells in spontaneous tolerance of liver allografts

It has been shown that rat liver allografts between certain inbred major histocompatibility complex (MHC) disparate strains are accepted spontaneously, and regulatory T cells (Tregs) have been suggested to play a role in the spontaneous liver tolerance. CD8⁺CD103⁺ T cells bear the phenotypes of effector cells, and they are implicated in allograft destruction. Here we provide evidence that CD8⁺CD103⁺ T cells possess regulatory function and may contribute to prevent liver allograft rejection. We show that the expression of CD103 in the CD8⁺ T cells was increased in spontaneous liver grafts tolerant recipients. We further show that CD8⁺CD103⁻ T cells can also upregulate the expression of CD103 and Foxp3 after stimulation with alloantigen or TGF-β in vitro, and the CD8⁺CD103⁺ T cells acquired regulatory properties. The suppressive function of the alloantigen or TGF-β conditioned CD8⁺CD103⁺ T cells was cell–cell contact dependent. These results imply that liver-specific factor(s) would be involved in the generation of CD8⁺CD103⁺ Tregs that contribute to spontaneous liver allografts tolerance.     

CD4+CD25+Foxp3+调节性T细胞与慢性乙型肝炎病毒感染的关系

目的 探讨调节性T细胞（Treg）在慢性乙型肝炎病毒感染中的免疫调节作用。方法 采用流式细胞术检测慢性活动性肝炎患者、乙型肝炎病毒携带者、乙型肝炎恢复者及健康对照者外周血CIN^＋T细胞中的CIN^＋CD25^＋Foxp3^＋调节性T细胞的比例。采用免疫组织化学法检测肝组织中Foxp3的表达。统计分析外周血中CIN^＋CD25^＋Foxp3^＋调节性T细胞的比例数与血清HBeAg和HBVDNA含量的关系。结果 外周血CIN^＋T细胞中CIN^＋CD25^＋Foxp3^＋调节性T细胞的比例数在慢性活动型肝炎组和乙型肝炎病毒携带组均高于健康对照组（P〈0．05），乙型肝炎病毒携带者组高于乙型肝炎恢复组（P〈0．05）。Foxp3在慢性活动型肝炎组肝组织中的阳性率和阳性细胞数均高于乙型肝炎病毒携带者组（P〈0．05）。外周血HBeAg（＋）患者Treg的比例数高于血清HBeAg（-）患者Treg的比例数（t=1．67．P〈0．05）。患者外周血CIN^＋CD25^＋Foxp3^＋Treg的比例数与血清HBV DNA含量呈正相关（r=0．56，P〈0．01）。结论 CIN^＋CD25^＋Foxp3^＋调节性T细胞可能与乙型肝炎病毒感染慢性化及乙型肝炎病毒的清除有关。     

原发性肾病综合征患儿CD4+CD2 5+调节性T细胞的变化及临床意义

目的 探讨原发性肾病综合征(PNS)患儿外周血淋巴细胞亚群及CD4+CD25+调节性T细胞的变化及其临床意义.方法 采用流式细胞术检测23例PNS(肾病组)患儿外周血T淋巴细胞亚群,同时检测17例健康儿童作为对照组.结果 PNS患儿外周血中CD4+CD25+淋巴细胞较正常对照组明显升高(P＜0.05).结论 体内异常的CD4+CD25+调节性T细胞水平参与了PNS的发病过程.PNS患儿存在细胞免疫功能紊乱,纠正或调节细胞免疫功能可应用于PNS的临床治疗.     

CD4+CD25+ regulatory T cells: a therapeutic target for liver diseases

Background: Regulatory T cells (T_regs) have been shown to play an important role in maintaining peripheral immune homeostasis by suppressing autoreactive and allergen-specific T cells and turning off the immune response after the pathogen has been cleared. However, in certain situations T_regs can impair effective immunity to some pathogens and tumour cells. Objective: To review the role of T_regs in liver pathology and to assess the potential to enhance or inhibit their function as applied to the treatment of liver disease. Methods: The literature was reviewed using standard indexing terms and incorporating publications up to and including those published in 2007. Results/conclusions: T_regs are therapeutic targets for modulation in autoimmune disease and may provide new opportunities for application to human liver conditions.     

CD4+CD25+T淋巴细胞在支气管哮喘患者外周血的表达及意义

目的 探讨外周血CD4+CD25+T淋巴细胞与支气管哮喘的关系.方法 用流式细胞术分别检测支气管哮喘患者34例(A组)和健康人25例(B组)外周血CD4+CD25+T淋巴细胞的百分率,采用ELISA法检测外周血血清白细胞介素10(IL-10)水平,用化学发光法检测外周血IgE水平,并分析CD4+CD25+T淋巴细胞与外周血IgE及IL-10水平的相关性.结果 A组外周血CD4+CD25+T淋巴细胞百分率显著低于B组[(5.90±1.10)%vs.(7.13±1.48)%](P<0.05),且与血IL-10呈正相关,与IgE呈负相关(P<0.05).结论 CD4+CD25+T淋巴细胞在支气管哮喘患者中高表达,与IL-10及IgE有一定的相关性,是参与支气管哮喘的发生、发展的重要T淋巴细胞.     

消癌平注射液联合化疗对中晚期肺癌患者CD4~+CD25~+FOXP3~+调节型T细胞的影响

目的探讨消癌平注射液联合化疗对中晚期肺癌患者CD4+CD25+FOXP3+调节型T细胞的影响。方法 71例Ⅲ～Ⅳ期非小细胞肺癌患者随机分为治疗组与对照组,治疗组36例采用化疗联合消癌平注射液;对照组35例单独采用化疗,两组化疗均采用GP方案。完成2周期后,通过流式细胞仪测定两组患者治疗前后外周血中CD4+CD25+FOXP3+调节型T细胞比例。结果治疗后治疗组患者CD4+FOXP3+T细胞占CD4+T细胞及CD4+CD25+T细胞的比例分别为(6.2±2.4)%和(27.6±6.0)%,而对照组分别为(8.2±0.5)%和(32.1±7.6)%,两组比较差异有统计学意义(P<0.05)。CD4+T细胞占总淋巴细胞的比例,治疗组较对照组明显提高(P<0.05)。结论消癌平注射液是一种较为理想的免疫增强剂,联合化疗可明显改善非小细胞肺癌患者的细胞免疫功能。     

CD4~+CD25~+调节性T细胞在类风湿性关节炎中的研究进展

CD4+CD25+调节性T细胞(CD4+CD25+Tregs)主要发挥抑制性免疫调节功能,在维持外周免疫耐受和预防自身免疫性疾病中发挥重要作用。该文综述了CD4+CD25+Tregs的特征、作用机制及其在类风湿性关节炎(RA)中的作用,以便进一步研究此群细胞的功能,为RA的防治提供新的思路。