过敏性紫癜患儿外周血CD4~+CD25~+CD127~-调节性T细胞表达及意义

目的探讨过敏性紫癜(schonlein-henoch purpura,HSP)患儿治疗前后外周血CD4+CD25+CD127-调节性T细胞的表达及其临床意义。方法采用免疫荧光流式细胞技术检测56例HSP患儿治疗前后及20例健康儿童外周血CD4+CD25+CD127-调节性T细胞的表达值。结果 HSP患儿治疗前CD4+CD25+CD127-调节性T细胞的表达值明显低于治疗后组和正常对照组(P<0.01),HSP治疗后组CD4+CD25+CD127-调节性T细胞表达值与正常对照组相比差异无统计学意义(P>0.05)。结论 HSP治疗前患儿外周血CD4+CD25+CD127-调节性T细胞的表达值较正常对照组明显降低,而在治疗有效后升高,说明调节性T细胞可能参与了HSP的发病和病情的发展。而CD4+CD25+CD127-Treg细胞水平的检测可作为对HSP患儿预后判断的一个有效指标。     

系统性红斑狼疮患者外周血CD4+CD25+CD127low/-调节性T细胞的检测及意义

目的 探讨用膜表面标志CD4+ CD25+ CD127low/-作为检测调节性T(Treg)细胞标记的可行性,并探讨其在系统性红斑狼疮(SLE)中的临床意义.方法 用流式细胞术检测SLE组及健康对照组外周血CD4+ CD25+ CD127low/-Treg细胞及CD4+ CD25+ FoxP3+ Treg细胞的比例,并分析两组CD4+ CD25+ CD127low/-Treg细胞与CD4+ CD25+ FoxP3+ Treg细胞比例之间的相关性.结果 SLE组外周血CD4+ CD25+ CD127low/-Treg细胞比例为(3.31±0.82)％CD4+ CD25+ FoxP3+ Treg细胞比例为(2.28±0.47)％,均显著低于健康对照组的(6.07±1.59)％和(5.01±1.09)％(P＜0.01).SLE组及健康对照组外周血CD4+ CD25+ CD127low/-Treg细胞比例与CD4+ CD25+FoxP3+ Treg细胞比例之间呈显著正相关(r=0.713、r=0.709,P＜0.01).结论 膜表面标志CD4+ CD25+ CD127low/-可以用来鉴定Treg细胞;SLE患者外周血CD4+ CD25+ CD127low/-Treg细胞的显著减少可能与SLE的发病有关.     

SLE患者外周血CD4+CD25+Treg细胞、IFN-γ及IL-17的表达及意义

目的 通过检测系统性红斑狼疮(SLE)患者外周血中CD4+ CD25+ Treg细胞计数、IFN-γ及IL-17的表达水平,探讨其在SLE发病中的作用.方法 选取SLE患者24例及健康对照者22例作为研究对象,并根据SLEDAI评分将SLE患者分为活动组和非活动组,运用流式细胞技术检测外周血中CD4+ CD25+ Treg细胞的水平变化;运用酶联免疫吸附法(ELISA)检测血清中IFN-γ及IL-17的含量,并进行相关性分析.结果 ①SLE患者外周血中CD4+ CD25+ Treg细胞占CD4+T细胞的比例显著低于对照组(P＜0.05);②SLE患者血清IFN-γ及IL-17水平较对照组明显升高(P＜0.05),且活动组血清IL-17水平较非活动组明显升高(P＜0.05);③SLE患者CD4+ CD25+ Treg细胞水平与SLEDAI评分呈负相关(r=-0.673,P＜0.05);血清IL-17水平与SLEDAI平分成正相关(r=0.467,P＜0.05).结论 SLE患者外周血中CD4+ CD25+ Treg细胞数量的减少和血清中IL-17、IFN-γ水平的升高,可能与SLE患者的细胞免疫紊乱有关,且CD4+ CD25+ Treg细胞数量减少或IL-17水平升高可引起疾病活动.     

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.     

广州地区健康学龄前儿童CD4+CD25nt/hiCD127loTreg细胞在外周血中的表达

目的 初步确定广州地区健康学龄前儿童外周血CD4+CD25nt/hiCD127lo/Treg细胞的表达,建立学龄前儿童CD4+CD25nt/hiCD127loTreg细胞的正常参考范围,为临床研究相关疾病提供参考依据.方法 收集150名3～6岁体检健康的学龄前儿童外周血,采用多色免疫荧光素标记和流式细胞术检测外周血中CD4+CD25nt/hiCD127loTreg细胞的表达.结果 健康学龄前儿童CD4+CD25nt/hiCD127loTreg细胞在外周血中约占CD4+T细胞的6.52％～6.97％,不同性别表达差异无显著性.结论 初步确定广州地区学龄前健康儿童CD4+CD25nt/hiCD127loTreg细胞在外周血中的正常范围,为临床研究儿童相关疾病提供参考依据.应用IL-7受体细胞膜表面标志物CD127联合检测外周血CD4+CD25nt/hiCD127loTreg细胞,能较好地反映完整的调节性Treg细胞.     

肺癌患者外周血CD4+CD25+调节性T细胞的检测及其临床意义

目的 探讨检测肺癌患者手术前后外周血CD4+CD25+调节性T细胞的临床意义.方法 原发性肺癌患者43例(肺癌组),健康体检者20例(健康对照组).应用流式细胞术检测其外周血CD4+CD25+调节性T细胞的水平.结果 肺癌组患者手术前外周血CD4+CD25+调节性T细胞表达高于健康对照组(P<0.01);肺癌组患者术后30 d时外周血CD4+CD25+调节性T细胞水平较术前明显下降(P<0.05).结论 肺癌患者外周血CD4+CD25+调节性T细胞的水平增高;肺癌患者术后外周血CD4+CD25+调节性T细胞的水平下调,提示术后机体抗肿瘤免疫功能有了一定程度恢复.     

CD4~+CD25~+调节性T细胞在晚期肺癌中的表达及临床意义

目的探讨CD4+CD25+调节性T细胞(Treg细胞)在晚期肺癌中的表达及其临床意义。方法应用流式细胞术分析30例晚期肺癌患者外周血CD4+CD25+Treg细胞表达水平,与10例健康人比较。结果晚期肺癌患者外周血CD4+CD25+Treg细胞数量增加(17.9±6.1)%,表达高于正常人(6.81±0.4)%(P<0.001)。肺癌患者外周血中Treg细胞数量与患者的病理类型无关(P>0.05),但与临床分期和组织学分化程度有关(P<0.01)。结论Treg细胞在肺癌患者中比率明显升高,并与临床进展有关。     

猪囊尾蚴病患者CD4~+ CD25~+调节性T细胞水平的变化

目的检测猪囊尾蚴病患者外周血中CD4+CD25+调节性T淋巴细胞及其FOXP3的表达情况,探讨CD4+CD25+调节性T淋巴细胞在猪囊尾蚴感染中的免疫调控作用及意义。方法采用流式细胞仪检测11例猪囊尾蚴病患者外周血中CD4+CD25+T淋巴细胞的含量,同时观察CD4+CD25+T细胞中表达FOXP3群体的百分含量。结果囊尾蚴病患者外周血中CD4+CD25+T淋巴细胞的百分含量为6.11%,较正常人(3.94%)明显升高(P<0.05);患者外周血中CD4+CD25+T细胞表达FOXP3的细胞百分含量为15.67%,与正常对照组(11.09%)有显著性差异(P<0.05)。结论猪囊尾蚴病患者外周血中CD4+CD25+T淋巴细胞的百分含量显著升高,表明CD4+CD25+调节性T细胞可能参与猪囊尾蚴感染的免疫抑制。     

匹多莫德对过敏性紫癜患儿外周血CD4+CD25+CD127- 调节性T细胞的影响

目的:观察匹多莫德对过敏性紫癜(HSP)患儿CD4+CD25+CD127- 调节性T细胞的影响。方法:选择80例HSP患儿随机分为匹多莫德治疗组和常规治疗组各40例,两组患儿均给予常规治疗,匹多莫德组加用匹多莫德颗粒口服。检测所有患儿治疗前及治疗2周后CD4+CD25+CD127-调节性T细胞比例及血清IL-10、IL-4 及INF-γ水平。结果:治疗后匹多莫德组CD4+CD25+CD127-调节性T细胞比例为(3.85±0.97)%,较常规治疗组(2.54±0.89)% 升高,差异有统计学意义(P<0.01)。治疗后匹多莫德组血清IL﹣10 和IL﹣4 水平为(29.58±8.42) pg/mL、(76.47±16.45) ng/L,较常规治疗组的(40.89±9.67) pg/mL、(96.52±18.79)ng/L 降低,差异均有统计学意义(P均<0.05);匹多莫德组血清INF鄄酌水平为(89.54±15.87)ng/L,较常规治疗组的(77.43±15.32)ng/L 升高,差异有统计学意义(P<0.05);匹多莫德组INF﹣γ/IL﹣4为1.17±0.43,较常规治疗组的0.81±0.32升高,差异有统计学意义(P<0.01)。结论:匹多莫德辅助治疗HSP患儿能提高CD4+CD25+CD127- 调节性T细胞比例,减轻炎症反应,纠正免疫失衡。     

慢性乙型肝炎患者外周血CD4+CD25+high调节性T细胞对免疫状态的影响

目的 探讨CD4 CD25 high调节性T(Tr)细胞在慢性乙型肝炎(CHB)患者外周血的变化及其对免疫状态的影响.方法 用流式细胞仪技术分析比较42例CHB患者及20例正常健康人外周血中CD4 CD25 highTr细胞,同时检测CD4 T细胞和CD8 T细胞.结果 与健康对照组相比,CHB组CD4 CD25 highTr细胞百分率显著升高(P＜0.05),而CD4 T细胞显著下降(P＜0.05);CD8 T细胞虽有增加趋势,但无差异(P＞0.05).CHB组CD4 CD25 highTr细胞百分率与HBV DNA载量有一定的关系,但与血清ALT水平无相关(P＞0.05).结论 CHB患者外周血CD4 CD25 highTr细胞升高可能是导致机体免疫功能下降的原因之一.     

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.