Detection of a CD4+CD8−CD3− cell subpopulation during the differentiation of cord blood CD34+ cells into T cells in vitro

Introduction:  

Umbilical cord blood contains relatively abundant primitive CD34⁺ hematopoietic progenitor cells which can differentiate into T lymphocytes ex vivo     

强直性脊柱炎患者外周血T细胞亚群及其CD3+CD69+、CD3+CD54+检测

强直性脊柱炎(ankylosing spondylitis,AS)是一种主要侵犯脊柱,并累及骶髂关节和周围关节,以慢性进行性炎症为主要表现的自身免疫性疾病.     

CD4+CD25+调节性T细胞研究进展

CD4+CD25+调节性T细胞是调节性T细胞的亚群之一,主要来源于胸腺,具有多种独特的特征,包括可识别自身抗原肽、分泌抑制性细胞因子等.其功能是通过抑制自身反应性T细胞的免疫反应、抑制传统T细胞的活化以及促进一些抑制性细胞因子的分泌等,在维持机体内环境的稳定、肿瘤免疫监测、诱导移植耐受以及自身免疫性疾病的发生中发挥重要作用.     

2-Gy whole-body irradiation significantly alters the balance of CD4+CD25? T effector cells and CD4+CD25+Foxp3+ T regulatory cells in mice

CD4⁺CD25⁺ T regulatory (Treg) cells are critical in inducing and maintaining immunological self-tolerance as well as transplant tolerance. The effect of low doses of whole-body irradiation (WBI) on CD4⁺CD25⁺Foxp3⁺ Treg cells has not been determined. The proportion, phenotypes and function of CD4⁺CD25⁺ Treg cells were investigated 0.5, 5 and 15 days after euthymic, thymectomized or allogeneic bone marrow transplanted C57BL/6 mice received 2-Gy γ-rays of WBI. The 2-Gy WBI significantly enhanced the ratios of CD4⁺CD25⁺ Treg cells and CD4⁺CD25⁺Foxp3⁺ Treg cells to CD4⁺ T cells in peripheral blood, lymph nodes, spleens and thymi of mice. The CD4⁺CD25⁺ Treg cells of the WBI-treated mice showed immunosuppressive activities on the immune response of CD4⁺CD25⁻ T effector cells to alloantigens or mitogens as efficiently as the control mice. Furthermore, 2-Gy γ-ray WBI significantly increased the percentage of CD4⁺CD25⁺Foxp3⁺ Treg cells in the periphery of either thymectomized mice or allogeneic bone marrow transplanted mice. The in vitro assay showed that ionizing irradiation induced less cell death in CD4⁺CD25⁺Foxp3⁺ Treg cells than in CD4⁺CD25⁻ T cells. Thus, a low dose of WBI could significantly enhance the level of functional CD4⁺CD25⁺Foxp3⁺ Treg cells in the periphery of naive or immunized mice. The enhanced proportion of CD4⁺CD25⁺Foxp3⁺ Treg cells in the periphery by a low dose of WBI may make hosts more susceptible to immune tolerance induction.     

Regulation of Con A – dependent cytokine production from CD4+ and CD8+ T lymphocytes by autosecretion of histamine

Objectives: Previously we have shown that both CD4+ T cells and CD8+ T cells produce histamine when activated with Con A. The aim of this study was to examine whether cytokine production by these cells is regulated by autosecretion of histamine.Materials: CD4+ and CD8+ T cells were separated from spleen cells of C57BL/6 mice and mice lacking the H1 receptor (H1R) or H2R, using anti – CD4+ – and anti – CD8+ – coupled magnetic beads, respectively.Results: Depletion of the H1R resulted in decreases in the release of IL-2 and IL-10 from both CD4+ and CD8+ cells and increases in the release of IL-4 from CD4+ T cells and IFN- from CD8+ cells. Mice lacking the H2R showed up – regulation of IFN- secretion from CD8+ cells and of IL-4 from CD4+ and CD8+ T cells. Release of IL-2 and IL-10 from CD4+ as well as CD8+ cells was down – regulated in these mice. Both CD4+ and CD8+ T cell fractions synthesized histamine, which was enhanced in the H1R - deficient CD8+ T cells. Treatment of the cells with -fluoromethyl-histidine, a specific inhibitor of HDC, or histaminase increased IFN- from CD8+ cells, whereas it had no appreciable effect on IL-4 secretion from CD4+ cells.Conclusions: These results suggest that cytokine production by CD4+ and CD8+ T lymphocytes is regulated by autosecretion of histamine.Received 4 July 2003; returned for revision 23 September 2003; returned for final revision 13 October 2003, accepted by M. Parnham 17 October 2003     

Higher Percentage of CD3+CD154+ T Lymphocytes Predicts Efficacy of TNF-α Inhibitors in Active Axial SpA Patients

The objective of this study was to evaluate which subtypes of T lymphocytes (CD3⁺CD28⁺ and CD3⁺CD154⁺) could predict clinical efficacy after TNF-α inhibitor treatment in active axial SpA patients. Patients who fulfilled Assessment of SpondyloArthritis international Society (ASAS) criteria for axial SpA had a BASDAI of ≥40 mm. All patients received TNF-α inhibitor treatment for 12 weeks. ASAS20 was used to evaluate the effect of the treatment at week 12. We detected the percentage of CD3⁺CD28⁺ and CD3⁺CD154⁺ T lymphocytes on lymphocyte cells in the peripheral blood in patients and healthy controls. We evaluated whether the percentage of the above subtypes of T lymphocytes could predict clinical efficacy by ROC curve analysis. Fifty-eight healthy controls and 74 active axial SpA patients were included. Mean age was 26.28?±?9.08 and 26.95?±?8.13 years for healthy controls and patients, respectively (p?=?0.767). The percentage of CD3⁺CD154⁺ T lymphocytes was significantly higher in axial SpA patients than in healthy controls (1.62?±?1.89 % vs 0.79?±?0.52 %, p?+CD154⁺ T lymphocytes was significantly higher in HLA-B27(⁺) patients than HLA-B27(^?) ones (HLA-B27⁺ vs HLA-B27^?:1.77?±?1.95 % vs 0.41?±?0.27 %, p?=?0.005). Compared with baseline, the percentage of CD3⁺CD154⁺ T lymphocytes significantly decreased to 0.87?±?0.49 % at week 12 (p?+CD154⁺ T lymphocytes could predict clinical efficacy of SpA patients with TNF-α inhibitor treatment (AUC?=?0.733, p?=?0.014). High percentage of CD3⁺CD154⁺ is over-expressed on lymphocytes in peripheral blood of active SpA patients and can be down-regulated by TNF-α inhibitor therapy. High-percentage of CD3⁺CD154⁺ T lymphocytes may predict clinical efficacy of TNF-α inhibitor treatment in active axial SpA patients.     

检测CD3+、CD4+、CD8+、CD4+/CD8细胞对慢性乙肝患者的临床意义

乙型肝炎病毒（HBV）是一种非细胞毒性病毒,当其进入机体后常可引起一系列复杂的免疫反应。而机体产生的免疫应答的强弱又与HBV感染所致的不同临床结果密切相关。而在这一系列免疫应答中,细胞免疫应答又是决定HBV感染机体后转归的最重要因素。而不同的T细胞亚群对病毒抗原的反应状态也不同,即对临床过程的轻重及转归的影响不同。本实验应用流式细胞仪对健康人、慢性乙型肝炎患者和乙肝肝硬化患者外周血中的测定CD4+、CD4+/CD8+、CD4+/CD8细胞的含量进行了检测计算,来探讨慢性乙型肝炎患者外周血中CD4+、CD4+/CD8+、CD4+/CD8细胞特点及其与肝病病情的关系。     

CD4+CD25+T细胞与移植免疫耐受

调节性T细胞是机体维持自身耐受的重要组成部分.CD4+CD25+T细胞以持续高表达CD25为特征,可通过细胞间直接接触或分泌TGF-β、IL-10来发挥抑制功能.它广泛参与自身免疫耐受、肿瘤免疫、移植免疫.现就其发育、特性、发挥功能的机制以及在移植免疫耐受中的作用和应用前景作一综述.     

CD4+CD25+T细胞在CD8+T细胞抗肿瘤免疫中的调节作用

实验旨在研究CD4^＋CD25^＋T细胞在CD8^＋T细胞抗肿瘤免疫中的调节作用。将小鼠脾脏中分离的单个核细胞分为两组．即去除CD4^＋CD25^＋T细胞组和未去除CD4^＋CD25^＋T细胞组,测定树突状细胞提呈的肿瘤抗原多肽刺激不同T细胞增殖活性、细胞因子IFN一1分泌,以及多肽特异性CD8^＋T细胞对同源性胃癌细胞株MFC的杀伤活性。结果显示预先去除未致敏T细胞中的CD4^＋CD25^＋T细胞,所诱导的特异性CD8^＋CTL对肿瘤细胞免疫应答增强,表现为反应性T细胞对树突状细胞提呈的肿瘤抗原多肽增殖反应增强,IFN-γ分泌量提高及CD8＋T细胞对MFC杀伤活性增强。这些结果表明。预先去除未致敏T细胞中的CD4^＋CD25^＋T细胞,肿瘤抗原多肽修饰的树突状细胞肿瘤疫苗效能可明显增加。CD4^＋CD25^＋T细胞在CD8^＋T细胞抗肿瘤免疫中起下调作用。     

CD4+CD25+T调节细胞的研究进展

CD4 CD25 TH细胞通过抗原特异性方式或细胞接触的方式抑制自身反应性T细胞的活化,能有效地维持自身免疫耐受,是调节自身反应性T细胞和防止自身免疫病发生的重要调节细胞。     

天然CD4+CD25+Treg细胞的研究进展

天然CD4+ CD25+ Treg细胞在针对自身抗原和外来抗原的免疫应答中起关键控制作用,其缺乏或功能性的缺陷将导致多重病理性的失调.本文就近年在其产生、作用机制以及与免疫耐受的诱导关系等方面的研究进展进行了综述.     

CD4+CD25+调节T细胞及其功能

CD4 CD25 T细胞亚群具有免疫调节功能,是机体内调节性T细胞(Teguolatory T cells,Tr)的主要类型,于1995年首次由Sakaguchi等提出.     

Differential sensitivity of naïve and subsets of memory CD4+ and CD8+ T cells to hydrogen peroxide-induced apoptosis

CD4+ and CD8+ memory T cells are identified into central and effector memory subsets, which are characterized by distinct homing patterns and functions. In this investigation, we show that na?ve and central memory CD4+ and CD8+ T cells are sensitive to hydrogen peroxide (H2O2)-induced apoptosis, whereas effector memory CD4+ and CD8+ T cells are relatively resistant to H2O2-induced apoptosis. Apoptosis in na?ve and central memory CD4+ and CD8+ is associated with the release of cytochrome c and activation of caspase-9 and caspase-3, upregulation of Bax and voltage-dependent anion channel (VDAC) expression, and decreased intracellular glutathione (GSH). In vitro GSH and a superoxide dismutase mimetic Mn(III) tetrakis (1-methyl-4-pyridyl) porphyrin inhibited H2O2-induced apoptosis in both na?ve and central memory CD4+ and CD8+ T cells. Furthermore, VDAC inhibitor 4,4'-diisothiocynostilbene-2,2'-disulfonic acid blocked H2O2-induced apoptosis. These data demonstrate that H2O2 induces apoptosis preferentially in human na?ve and central memory CD4+ and CD8+ T cells via the mitochondrial pathway by regulating intracellular GSH and the expression of Bax and VDAC.     

TGF-β1诱导CD4+CD25-T细胞分化为CD4+CD25+调节性T细胞

目的 研究TGF-β1诱导CD4 CD25 调节性T细胞分化的能力及其相关机理.方法 ①利用丝裂霉素处理的Balb/C裸鼠脾细胞(同种抗原)刺激C57BL/6小鼠T细胞,同时给予TGF-β1予以干预(按TGF-β1剂量不同设立4组:对照组,低浓度组,中浓度组和高浓度组),共同培养5天后,用流式细胞仪检测CD4 CD25 T细胞比例;同时用RT-PCR检测培养细胞中Foxp3的表达水平.②在第一部分实验基础上,通过MACS分离获取C57BL/6小鼠CD4 CD25-T细胞,用同种抗原刺激后,给予TGF-β1干预,培养5 d后分离CD4 CD25 T细胞,利用混合淋巴细胞培养系统检测其抑制淋巴细胞增殖的能力.结果 T细胞经同种抗原刺激后,在中、高浓度TGF-β1诱导下CD4 CD25 T细胞比例均明显升高,Foxp3的表达水平也相应增加,与对照组相比有显著性差异(P＜0.05).TGF-β1可直接诱导CD4 CD25-T细胞转化为CD4 CD25 T细胞,转化后的CD4 CD25 T细胞可有效抑制淋巴细胞增殖.结论 TGF-β1可诱导CD4 CD25-T细胞转化为CD4 CD25 Treg,促进其表达Foxp3,并能够抑制淋巴细胞增殖.     

非小细胞肺癌患者外周血CD11b~+CD33~+CD15~+CD14~-及CD11b~+CD33~+CD15~-CD14~+髓系细胞的比例变化及临床意义

髓源性抑制细胞(myeloid-derived suppressor cell,MDSC)是一群具有免疫抑制作用的细胞,其在外周血中的表达规律与非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的外周免疫逃逸及临床分期密切相关。本研究纳入116例NSCLC患者及30例健康者,流式细胞术检测外周血PBMC中MDSC、调节性T细胞(regulatory T cell,Treg)的比例,研究其与临床分期、病理及免疫的相关规律。结果显示:与健康组比较,粒系髓源性抑制细胞(G-MDSC)、单核系髓源性抑制细胞(M-MDSC)在NSCLC外周血中比例升高,具有统计学意义(P<0.05);G-MDSC、病理类型与临床分期具有相关性(n=116,r=0.330,P<0.001;n=116,r=0.441,P<0.001),而M-MDSC、Treg与临床分期之间未发现相关性(n=116,r=-0.053,P=0.558;n=116,r=0.173,P=0.052);G-MDSC与Treg具有相关性(n=116,r=0.343,P<0.001)且与病理类型也呈现正相关(r=0.333,P<0.001);而M-MDSC与Treg未发现相关性(r=0.122,P=0.174),与病理类型不具有相关性(r=-0.143,P=0.109)。研究表明NSCLC患者外周血PBMC中MDSC比例升高,其中G-MDSC与NSCLC临床分期及Treg呈正相关,调控MDSC的表达有望成为防治肺癌发生及术后复发与转移的新策略[临床研究国际注册号NCT02603003]。     

CD200 与CD4+ CD25+ CD127low Treg 在强直性脊柱炎中的意义

目的:研究强直性脊柱炎(AS)患者外周血淋巴细胞中CD200的表达水平,分析CD200及Treg与IL-6、SAA的相关性,探讨CD200与AS免疫功能的关系以及诊断效能,为辅助AS的诊断、免疫评估提供帮助。方法:从74例AS患者和50例健康志愿者中收集外周血。流式细胞术检测淋巴细胞亚群、Treg的百分率及CD200的表达率,并用ELISA法检测血清中CD200的浓度,上转发光法测定IL-6的浓度,胶乳免疫比浊法测定SAA的浓度。结果:与对照组相比,AS活动组CD4~+T细胞、CD19~+B细胞的百分率均增高(P0.05),CD8~+T细胞、Treg的百分率均减低(P0.05),CD3~+T细胞的百分率无明显差异;稳定组CD4~+T细胞、CD8~+T细胞、CD19~+B细胞、Treg的百分率与对照组均无显著差异。与对照组比较,活动组(CD3~+、CD4~+、CD8~+)T细胞中CD200的表达率和血清中CD200的浓度以及稳定组(CD3~+、CD4~+)T细胞中CD200的表达率和血清中CD200的浓度均减低(P0.05);而稳定组CD8~+T细胞、CD19~+B细胞中CD200的表达率及活动组CD19~+B细胞中CD200的表达率无显著差异。与对照组比较,活动组血清中IL-6、SAA的浓度明显升高(P0.05);而稳定组IL-6的浓度轻度升高(P0.05),SAA的浓度无差异。活动组CD3~+T淋巴细胞中CD200的表达率与Treg呈正相关,与IL-6呈负相关,但与SAA不相关;Treg的百分率与IL-6负相关,与SAA不相关。CD200对AS的诊断优于Treg、SAA,而与IL-6的诊断效能相当。结论:AS活动期患者存在淋巴细胞亚群紊乱,CD200和Treg的表达率降低,两者可能参与了AS的发病,为辅助AS的诊断和免疫功能的判断提供理论依据。