系统性红斑狼疮患者外周血CD4~+调节性T细胞CD25~+和FOXP3~+的表达及意义

目的探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中CD4+T细胞CD25和FOXP3的表达及其临床意义。方法流式细胞术检测20例SLE患者(9例为狼疮性肾炎)及10例健康对照组PBMC中CD4+T细胞中CD25及Foxp3的表达。结果 SLE患者CD4+CD25highT细胞中FOXP3+T细胞的表达明显低于健康对照组(P<0.05),与SLE疾病活动指数SLEDAI积分明显负相关(r=-0.514,P<0.01),与补体C4正相关(r=0.362,P<0.05),与ESR及CRP负相关(r分别=-0.23、-0.216,P<0.05),与外周血免疫球蛋白IgG、IgA、IgM及补体C3水平无明显相关性。将SLE患者分为狼疮性肾炎(LN)组及非肾损(SLE-NR)组,两组CD4+CD25highT细胞及CD4+CD25lowT细胞中FOXP3+T细胞的表达均明显低于健康对照组(P<0.05);LN组CD4+T细胞中CD25high及CD25low的表达均较SLE-NR组增高(P<0.05)。结论 SLE患者外周血中CD4+CD25high T细胞中FOXP3+的表达水平明显降低,且与疾病活动性相关,而LN患者代偿增高的无FOXP3表达的CD4+CD25+T细胞可能与其发病机制有关。     

Expression and significance of CD4+ CD25+ FOXP3+ regulatory T cells in patients with systemic lupus erythematosus

目的 探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中CD4+T细胞CD25和FOXP3的表达及其临床意义.方法 流式细胞术检测20例SLE患者(9例为狼疮性肾炎)及10例健康对照组PBMC中CD4+T细胞中CD25及Foxp3的表达.结果 SLE患者CD4+CD25highT细胞中FOXP3+T细胞的表达明显低于健康对照组(P＜0.05),与SLE疾病活动指数SLEDAI积分明显负相关(r=-0.514,P＜0.01),与补体CA正相关(r=0.362,P＜0.05),与ESR及CRP负相关(r分别=-0.23、-0.216,P＜0.05),与外周血免疫球蛋白IgG、IgA、IgM及补体C3水平无明显相关性.将SLE患者分为狼疮性肾炎(LN)组及非肾损(SLE-NR)组,两组CD4+ CD25high T细胞及CD4+ CD25lowT细胞中FOXP3+T细胞的表达均明显低于健康对照组(P＜0.05); LN组CD4+T细胞中CD25high及CD25low的表达均较SLE-NR组增高(P＜0.05).结论 SLE患者外周血中CD4+ CD25highT细胞中FOXP3+的表达水平明显降低,且与疾病活动性相关,而LN患者代偿增高的无FOXP3表达的CD+CD25+T细胞可能与其发病机制有关.     

系统性红斑狼疮患者T细胞亚群的变化

目的:探讨T细胞亚群在系统性红斑狼疮(SLE)发病机制中的作用及其能否作为SLE病情评估和指导治疗的参考指标.方法:收集SLE高活动期患者40例,低活动期患者26例,稳定期患者15例,健康查体者30例.采用流式细胞术检测外周血样本的CD4+、CD8+细胞百分率及CD4+/CD8+比值;记录SLE患者的临床表现、疾病活动性指数(SLEDAI)评分、一般实验室检查以及血清免疫学指标,并对所得数据进行比较和相关性分析.结果:SLE患者不同活动期之间及和正常对照组之间外周血T细胞亚群存在差异.高活动期SLE患者CD4+/CD8+比值与SLDAI评分及尿蛋白负相关,与补体C3呈正相关;高活动期SLE患者经糖皮质激素和免疫抑制剂等治疗2周后,SLEDAI评分下降,T细胞亚群较治疗前无明显变化.结论:T细胞亚群SLE的发病机制中具有一定作用,并与疾病的严重程度有关,但不能作为短期内治疗是否有效的参考指标.     

CD4+CD25+调节性T细胞在再生障碍性贫血中的表达及意义

目的:探讨再生障碍性贫血(再障)患者治疗前、后外周血T淋巴细胞亚群、CD4+CD25+T细胞的变化及其临床意义.方法:采用流式细胞术检测23例再障患者治疗前、后外周血T淋巴细胞亚群、CD4+CD25+T细胞的比例,并与健康对照组进行比较.结果:(1)再障患者治疗前CD4+,CD4+/CD8+,CD4+CD25+明显低于健康对照组(P＜0.01),CD8+细胞则明显升高(P＜0.05);重型再障组与慢性再障组比较CD4+CD25+降低,但差异不显著(P＞0.05),CD3+,CD4+,CD4+/CD8+,CD4+CD25+无统计学意义.(2)再障患者治疗后与治疗前相比,CD4+,CD4+/CD8+,CD4+CD25+明显升高(P＜0.01).(3)再障患者治疗有效者与无效者比较CD4+,CD4+/CDs+,CD4+CD25+明显升高(P＜0.01).结论:再障患者存在T淋巴细胞亚群的失调及CD4+CD25+T淋巴细胞的异常表达,且检测外周血T淋巴细胞亚群、CD4+CD25+T细胞有助于了解再障患者的免疫状态、疗效评价及预后判断.     

SLE患者外周血T淋巴细胞亚群和免疫球蛋白及补体水平的变化研究

目的探讨SLE患者外周血T淋巴细胞亚群、免疫球蛋白及补体水平的变化。方法采用多参数三色免疫荧光标记流式细胞术结合血液分析仪对136例SLE患者进行外周血T淋巴细胞亚群的测定,同时应用双光径免疫浊度分析仪检测其血清免疫球蛋白及补体水平。结果与正常对照组比较,SLE患者组外周血CD3+T淋巴细胞、CD4+T淋巴细胞百分比明显降低(P<0.05),CD8+T淋巴细胞百分比明显升高(P<0.05),CD4+/CD8+比值明显降低(P<0.01);SLE患者组血清IgG显著升高(P<0.01),IgA也升高(P<0.05),而IgM与对照组相比较并无明显差异(P>0.05);SLE患者组C3、C4均显著下降(P<0.01)。结论与正常对照组比较,SLE患者T淋巴细胞亚群、免疫球蛋白及补体有明显变化。     

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.     

探析系统性红斑狼疮患者CD4+CD25+调节性T细胞的变化

本文首先使用流式细胞术检测xx例SLE患者及XX名正常人,对照两个实验组的外周血中CD4+CD25+调节性T细胞和CD4+CD25+T细胞占CD4+T细胞的比率,深入分析CD4+CD25+调节性T细胞与SLE之间的关系,通过对照实现发现活动性SLE患者外周血中CD4+CD25+调节性T细胞及CD4+CD25+T细胞比例低于正常人,而且与红斑狼疮的SLEDAI指数呈现负相关等实验结果。希望通过本文的研究能够更加清楚的了解系统性红斑狼疮患者CD4+CD25+调节性T细胞的变化以及与SLE发病之间的关系,从而为后期的临床研究治疗提供参考。     

CD19+CD24hiCD38hi B细胞亚群在系统性红斑狼疮患者外周血中的表达

目的探讨CD19+CD24hi CD38hi B细胞亚群在系统性红斑狼疮(SLE)患者外周血中的表达及临床意义。方法流式细胞术检测SLE患者(S组,36例)和正常对照组(C组,20例)CD19+CD24hi CD38hi、CD19+CD24int CD38int、CD19+CD24hi CD38-B细胞亚群百分比水平。S组中,20例为SLE活动(S1组),16例为SLE稳定(S2组)。结果 S组CD19+CD24hi CD38hi B细胞亚群百分比显著高于C组[(10.93±6.30)%vs.(7.51±2.73)%](P<0.05),且S1组高于S2组(P<0.05)。S组CD19+CD24hi CD38-B细胞亚群百分比显著低于C组[(15.52±9.54)%vs.(26.46±11.07)%](P<0.05)。S组CD19+CD24hi CD38hi B细胞亚群表达水平与SLE疾病活动指数(SLEDAI)积分正相关(P<0.05)。结论 CD19+CD24hi CD38hi B细胞亚群对效应T细胞分泌促炎症因子缺乏抑制功能与其存在功能缺陷有关,而非因其数量不足。CD19+CD24hi CD38hi B细胞亚群在SLE患者外周血中的表达水平可能与疾病活动相关。     

外周血T淋巴细胞对乙肝病毒相关慢加急性肝衰竭患者预后的影响

目的 探讨乙肝病毒相关慢加急性肝衰竭(HBV-ACLF)患者外周血T淋巴细胞亚群比例对其预后的影响.方法 选择HBV-ACLF患者43例,随访观察3个月后患者的存活率,应用流式细胞仪测定外周血CD3+、CD4+和CD8+T淋巴细胞及CD4+ CD25+ Treg细胞的百分比,并计算CD4+和CD8+T淋巴细胞比值,分析T淋巴细胞亚群对乙肝病毒相关慢加急性肝衰竭患者预后的影响,选择门诊同期健康体检者20例为健康对照组.结果 3个月后,43例HBV-ACLF患者存活26例,死亡17例.与肝衰竭存活组相比,肝衰竭死亡组CD3+细胞百分率(22.96±20.59)％、CD8+细胞百分率(31.63±12.69)％均低于存活组(37.89±17.36)％和(36.52±9.75)％,而CD4+细胞百分率(55.15±14.23)％、CD4+/CD8+(1.77±1.38)高于存活组(48.51±13.35)％、(1.32±0.68),均差异有统计学意义(P＜0.05),但两组之间CD4+ CD25+ Treg细胞比例差异无统计学意义.与健康对照组相比,肝衰竭死亡组患者外周血CD3+、CD8+T淋巴细胞及CD4+ CD25+ Treg细胞比例下降,CD4+T淋巴细胞、CD4+/CD8+升高,肝衰竭存活组患者CD3+T淋巴细胞及CD4+ CD25+ Treg细胞百分比下降,均差异有统计学意义(P＜0.05).结论 T淋巴细胞亚群比例的变化在一定程度上可以预测HBV-ACLF患者的预后,外周血T淋巴细胞CD3+、CD8+、CD4+ CD25+ Treg比例下降的程度越重,预后可能越差.     

危重型甲型H1N1流感患者外周血白细胞及T细胞亚群的变化及其临床意义

目的 探讨危重型甲型H1N1流感患者外周血白细胞和T淋巴细胞亚群的变化及其临床意义.方法 检测18例危重型甲型H1N1流感患者(危重组),18例普通型H1N1流感患者(普通组)和18例健康人(对照组)外周血白细胞计数及分类,流式细胞术测上述患者外周血T淋巴细胞各亚群CD3+、CD4+、CD8+、CD8+CD28+、CD8+CD28-、CD4+CD25+high的百分比.结果 与对照组比较,甲型H1N1病毒感染患者,尤其是危重型组,淋巴细胞绝对数及百分比显著下降,且外周血T淋巴细胞各亚群CD3+、CD4+、CD8+、CD8+CD28+、CD4+CD25+high百分比明显降低,而CD8+CD28-百分比明显增高(P<0.05).结论 甲型H1N1流感病毒感染损伤患者淋巴细胞和细胞免疫功能.外周血淋巴细胞及T淋巴细胞各亚群的百分比改变可反映甲型H1N1流感患者病情的严重程度.     

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".     

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.     

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.     

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.     

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.     

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.