IKK2dn转染树突状细胞诱导产生的CD4+CD25-T细胞的筛选及鉴定

目的 探讨从转染IKK2dn并负载供者抗原的未成熟树突状细胞(imDC)诱导产生的调节性T细胞(Treg)中筛选CD4+ CD25 - Treg的方法,并进行鉴定.方法 Lewis大鼠骨髓源性imDC,转染IKK2dn后负载供者BN大鼠抗原,与Lewis大鼠T细胞进行体外混合淋巴细胞反应(MLR)诱导产生Treg,用免疫磁珠法(MACS)筛选出CD4+ CD25 -T细胞,流式细胞仪(FCM)检测细胞纯度.加入CD4+ CD25 -T细胞行再次MLR检测其抑制T细胞增殖的作用.结果 经MACS筛选,CD4+ CD25 -T细胞纯度为(95.78±1.25)％.再次MLR结果显示CD4+ CD25 -T细胞组的吸光度值为(0.106±0.006),低于BN抗原组(0.189±0.007)、Adv0-CD4+T细胞组(0.419±0.014)及第三方供者抗原组(0.200±0.008),差异有统计学意义(P＜0.05).结论 转染IKK2dn并负载抗原的imDC诱导产生的Treg,经MACS筛选可以获得高纯度的CD4+CD25-T细胞,对同种T细胞增殖具有针对供者的特异性免疫抑制作用.     

信号转导和转录激活因子5在小鼠CD4+CD25+调节性T细胞中的表达

目的:观察信号转导和转录激活因子5(STAT5)在小鼠CD4+CD25+调节性T细胞中的表达情况.方法:免疫磁珠法分离C57BL/6J小鼠脾脏中的CD4+CD25+调节性T细胞,共聚焦荧光法检测细胞内STAT5的分布并进行初步定位,进一步采用Western blot技术从蛋白水平检测细胞内STAT5的表达.结果:激光共...     

Isolation, expansion and functional assessment of CD4+CD25+FoxP3+ regulatory T cells and Tr1 cells from uremic patients awaiting kidney transplantation

Background

The immunosuppressive properties of regulatory T cells have emerged as an attractive tool for the development of immunotherapies in various disease contexts, e.g. to treat transplantation induced immune reactions. This paper focuses on the process of obtaining and functionally characterizing CD4+CD25+FoxP3+ regulatory T cells and Tr1 cells from uremic patients awaiting kidney transplantation.

Methods

From October 2010 to March 2011 uremic patients awaiting living donor kidney transplantation, and their corresponding kidney donors, were enrolled in the study. A total of seven pairs were included. Isolation of CD4+CD25+FoxP3+ regulatory T cells was performed by magnetic activated cell sorting of peripheral blood mononuclear cells obtained from the uremic patients. Donor specific Tr1 cells were differentiated by repetitive stimulation of immature CD4+ T cells with immature dendritic cells, with the T cells coming from the future kidney recipients and the dendritic cells from the corresponding kidney donors. Cells were then expanded and functionally characterized by the one-way mixed leukocyte reaction and assessment of IL-10 production. Phenotypic analysis was performed by flow cytometry.

Results

The fraction of CD4+CD25+FoxP3+ regulatory T cells after expansion varied from 39.1 to 50.4% and the cells retained their ability to substantially suppress the mixed leukocyte reaction in all but one patient (3.8-19.2% of the baseline stimulated leukocyte activity, p < 0.05). Tr1 cells were successfully differentiated from all but one patient and produced high levels of IL-10 when stimulated with immature dendritic cells (1,275-11,038% of the baseline IL-10 secretion, p < 0.05).

Conclusion

It is practically feasible to obtain and subsequently expand CD4+CD25+FoxP3+ regulatory T cells and Tr1 cells from uremic patients without loss of function as assessed by in vitro analyses. This forms a base for adoptive regulatory T cell therapy in the setting of living donor kidney transplantation.     

CD4+CD25+调节性T细胞对小鼠肝脏移植自发性免疫耐受的影响

目的 研究CD4+CD25+调节性T细胞在诱导自发性肝脏免疫耐受中的作用.方法 向受体和供体注射抗CD25抗体(PC61)后进行小鼠原位肝脏移植,观测其生存时间.术后20～30 d切取移植肝脏行HE染色,同时观察CD4+CD25+T细胞对CD4+T细胞和CD8+T细胞功能的影响.结果 去除受体而不是供体小鼠的CD4+CD25+T细胞可以导致肝移植排斥反应.而且,去除CD4+CD25+T细胞使移植物的白细胞浸润明显增多,组织损伤加重.同时,去除CD4+CD25+T细胞导致CD4+T细胞的增殖活性和CD8+T细胞的细胞毒活性明显增强.结论 受体来源的CD4+CD25+调节性T细胞在小鼠肝脏移植免疫耐受诱导中起重要作用.

Abstract：

Objective To examine the contribution of CD4+ CD25+ regulatory T cells to liver transplant tolerance. Methods After injection of anti-CD25 monoclonal antibody (mAb, PC61), mouse orthotopic liver transplantation was performed and survivals were determined. The paraffin-embedded sections of hepatic allografts were cut and stained with hematoxylin and eosin (HE). Furthermore, the effect of CD4+ CD25+ regulatory T cells on proliferative response of CD4+ T cells and cytotoxicity of CD8+ T cells was examined by depleting these regulatory T cells. Results Depletion of these cells in the recipients but not in the donors before liver transplantation caused rejection. Histological analyses of hepatic allografts with PC61 treatment showed extensive leukocyte infiltration and tissue destruction, whereas those in the control group showed minimal changes. Moreover, elimination of CD4+CD25+ T cells resulted in the enhancement of both proliferative response of CD4+ T cells and cytotoxicity of CD8+ T cells against donor-type alloantigen. Conclusions These results suggest that CD4+CD25+ regulatory T cells were important for tolerance induction to hepatic allografts.     

Immune regulation by regulatory T cells: implications for transplantation

The induction of antigen-specific T cell tolerance and its maintenance in the periphery are critical for the immune system to prevent autoaggressive immune responses. Our current state of knowledge about the immunoregulatory mechanisms responsible for T cell tolerance in the periphery offers new possibilities for immunomodulation to prevent transplant rejection as well as to diminish autoimmune reaction or chronic allergy. There is growing evidence that dendritic cells, besides their well-known T cell stimulatory functions, also maintain and regulate T cell tolerance in the periphery. This control function is exerted by certain maturation stages and subsets of dendritic cells, and can be further influenced and modulated by immunoregulatory cytokines and drugs. The regulatory functions of dendritic cells include the induction of T cell anergy, of T cells with regulatory properties and of T cells that produce immunosuppressive cytokines such as IL-10 or TGF-beta. Additionally, distinct subsets of resident regulatory T cells generated in the thymus play a central role in maintenance of peripheral tolerance by active suppression of effector T cell populations. These CD4(+)CD25(+) regulatory T cells inhibit a variety of autoimmune and inflammatory diseases and they are also efficient in the suppression of alloantigen responses. This review summarises the current knowledge regarding the immunoregulatory role of dendritic cells and the functional activities of resident regulatory T cells as guardians for peripheral T cell tolerance.     

慢性乙型肝炎患者外周血调节性T细胞对树突状细胞功能的抑制作用

目的研究慢性乙型肝炎（CHB）患者外周血CD4＋CD25＋调节性T细胞（Treg）对树突状细胞（DCs）免疫功能的抑制作用,探讨治疗CHB的新方法。方法采用密度梯度离心法获得CHB患者和健康对照组（NC组）外周血单个核细胞（PBMC）;部分PBMC体外诱导培养获得DCs,部分PBMCs用特异性免疫磁珠分选获得CD4＋CD25＋Treg和CD4＋CD25-T细胞;不同来源的DCs和正常对照组CD4＋CD25-T细胞混合为反应细胞,将不同来源和不同比例的Treg分别加入到反应细胞中培养3 d,MTT法检测Treg抑制DCs的抑制指数（SI）,并在培养DCs的不同时间加入Treg,应用流式细胞术检测DCs表面共刺激分子CD80和HLA-DR的表达。结果来源于CHB患者及NC组的Treg均可抑制DCs的免疫作用,来源于CHB患者Treg抑制DCs能力显著高于NC,差异具有统计学意义（P ＜0.01）。不同比例的Treg均可抑制DCs的免疫功能,随着Treg比例的增高抑制作用也越明显,抑制指数亦越高。在DCs培养的不同时间加入相同比例的Treg,发现Treg对DCs表面分子CD80、HLA-DR的表达均有抑制作用,与对照组相比,差异具有统计学意义（P＜0.01）,同时发现加入Treg的时间越早,DCs表面分子表达降低越明显。结论 CHB患者Treg可显著抑制DCs免疫功能且呈时间和量的依赖,抑制DCs表面分子CD80和HLA-DR表达,可能是Treg抑制DC免疫功能的机制之一。     

体外富集Treg细胞对CTL杀伤MNK45胃癌细胞活性的影响及机制研究

目的体外分析CD4 CD25 Treg(regulatory T cells)细胞对CD8 CTL(cytotoxic Tlym-phocytes)细胞杀伤MNK45胃癌细胞株活性的影响及机制。方法通过T细胞纯化柱分选小鼠脾细胞悬液T淋巴细胞,以MACS单阳性和双阳性分选法分别富集CD4 CD25 Treg细胞和CD8 CTL;通过不同浓度CD8 CTL与MNK45胃癌细胞混合培养,筛选有效杀瘤体外培养体系。通过Trans Well分隔培养和混合培养的方式向CTL与MNK45胃癌细胞株组成的体外抗肿瘤培养体系中加入CD4 CD25 Treg细胞,以瑞氏染色显微计数的方法观察Treg细胞对CTL杀伤MNK45胃癌细胞活性的影响,结合ELISA法检测各体系穿孔素浓度,分析影响活性的细胞及免疫学机制。结果质量浓度为2.0×105/ml的CTL与1.0×105/ml MNK45胃癌细胞混合培养,CTL能对MNK45胃癌细胞形成明显的杀伤作用(P<0.01);在此培养体系中以混合培养的方式加入质量浓度为5.0×104/ml CD4 CD25 Treg细胞即可明显抑制培养体系中CTL对肿瘤细胞的杀伤作用;如以Trans Well分隔培养的方式将CD4 CD25 Treg细胞与抗肿瘤培养体系分隔培养,达到同样抑制效能(实验组死亡肿瘤细胞百分率与对照组比较,P<0.05)的CD4 CD25 Treg细胞浓度须≥2.0×105/ml。结论CD4 CD25 Treg细胞能够对CTL杀伤MNK45胃癌细胞株的活性形成明显抑制,在Treg细胞与CTL作用的相应细胞和免疫学机制中,细胞接触机制占主导地位。     

RNA干扰阻断OX40/OX40L共刺激通路对CD4+CD25+T调节细胞的影响

目的 探讨慢病毒介导的RNA干扰技术诱导小鼠树突状细胞OX40L基因沉默对调节性T细胞的影响. 方法 设计针对小鼠OX40L基因的RNA于扰序列,通过外源筛靶筛选出干扰效果最佳的序列.以293T为包装细胞,制备含OX40L的siRNA序列的慢病毒载体OX40L-RNAi-LV和阴性对照载体NC-GFP-LV.采用磁式分选器分离培养骨髓来源的小鼠树突状细胞(DCs),以MOI为25进行转染,分别将转染OX40L-RNAi-LV(实验组)、转染NC-GFP-LV(阴性对照组)和未转染(空白对照组)的DCs与磁式分选器分选得到的CD4+CD25+T调节细胞共培养,6 d后通过流式细胞仪检测T调节细胞的增殖和凋亡情况. 结果外源筛靶筛选出干扰效果最佳的RNA干扰序列(靶序列为GCTCATACAAGAATGAGTA),OX40L蛋白表达的抑制率为73.1%.感染复数为25时,慢病毒载体感染DCs的效率为86.4%.DCs与CD4+CD25+T调节细胞共培养后,实验组的凋亡细胞比例为8.7%,显著低于阴性对照组(20.1%)和空白对照组(19.8%),F=244.22,P=0.000;而增殖细胞前体频率为38.3%,明显高于阴性对照组(24.5%)和空白对照组(22.9%),F=95.40,P=0.000.结论 小鼠OX40L的siRNA慢病毒载体可以有效降低树突状细胞OX40L的表达,对体外培养的CD4+CD25+T调节细胞具有显著地促进增殖、减少凋亡的作用.     

Immune regulation by CD4+CD25+ regulatory T cells: implications for transplantation tolerance

In transplantation research, the achievement of life-long tolerance for the graft without the need for immunosuppressive drugs, is a major goal. In the immune system various mechanisms are in place that help to prevent unwanted immunity. These mechanisms of peripheral tolerance include deletion, anergy, ignorance and suppression. In the last decade it has been demonstrated convincingly that a naturally occurring subset of CD4+ T cells, the so-called CD4+CD25+ regulatory T cells, play a key role in the suppression/regulation of immune responses. These cells have been shown to exist in mice, rats and humans, and can be found in thymus, peripheral blood, lymphoid organs and at sites of inflammation. CD4+CD25+ regulatory T cells can down-regulate the immune response by affecting T cell responses, antibody production, cytokine secretion and antigen-presenting cells. CD4+CD25+ regulatory T cells are generated in the thymus, but importantly recent evidence suggests that they can also be generated in the periphery. This latter finding is of particular importance for transplantation immunology, since it suggests that specific manipulation or induction of these cells is achievable in vivo. Here we review the recent developments on the CD4+CD25+ regulatory T cells and we discuss the potential use of these cells in transplantation immunology.     

血栓闭塞性脉管炎与调节性T细胞的关系

【摘要】〓目的〓探讨CD4+CD25+调节性T细胞(Treg)与血栓闭塞性脉管炎(TAO)的关系。方法〓收集35例TAO患者,以35例动脉硬化性闭塞症(ASO)患者作为阳性对照组,35例健康人作为阴性对照组。采用流式细胞术检测外周血Treg及其转化生长因子β1(TGF-β1)及白介素10(IL-10)百分含量;采用ELISA法检测胞浆型抗中性粒细胞胞浆抗体(CANCA)和核周型抗中性粒细胞胞浆抗体(PANCA),并分析其相关性。结果〓与正常对照组比较,TAO组患者Treg及TGF-β1均显著升高(P<0.05),而ASO组患者Treg及TGF-β1则显著低于正常对照组,三组间Treg及TGF-β1两两差异具有统计学意义(P<0.05)。与Treg及TGF-β1不同,TAO及ASO组患者IL-10组间无统计学差异,且均显著低于正常对照组(P<0.0)。与正常对照组比较,TAO及ASO组患者CANCA及PANCA均呈不同程度升高,且TAO患者CANCA及PANCA均显著高于正常对照组,而ASO组患者与正常对照组之间并无统计学差异(P<0.05)。Pearson相关性分析显示Treg、TGF-β1及IL-10三者任意一者均与CANCA及PANCA呈显著正相关性(P<0.05)。结论〓Treg及TGF-β1升高可能参与TAO的发病机制。     

调节性CD4~+CD25~+T细胞的分离纯化及免疫功能研究

目的 制备调节性CD~+ CD25~+T细胞(Treg)分析其免疫功能,诱导局部免疫耐受防治同种异体复合组织移植(CTA)排斥反应.方法 采用免疫磁珠法(MACS)从雄性大鼠脾脏细胞分离CD4~+CD25~+Treg(1×10~6),2%锥虫蓝染色检测活性、流式细胞术分析其纯度,在5 mg/L抗CD3的刺激下观察其反应性、增殖及其与200 U/ml细胞介素(IL)-2的关系.结果 从8只雄性大鼠脾脏分选出的CD4~+CD25~+Treg活性平均为(97.90±0.36)%及纯度为(96.05±0.41)%,CD3刺激呈低反应,按比例培养抑制率为89%,IL-2可使CD4~+CD25~-抑制逆转.结论 MACS能快速分选出较高纯度的CD4~+CD25~+Treg,并且活性良好在体外具有免疫无能及免疫抑制作用,能满足动物CTA排斥反应研究的需要.     

CD4+CD25+调节性T细胞/辅助性T细胞17在脓毒症大鼠中的作用

目的 观察CD4+ CD25+调节T细胞(Treg)/辅助性T细胞17(Th17)细胞在脓毒症大鼠炎性免疫反应中的作用.方法 110只雄性SD大鼠随机分为正常对照组、假手术组、脓毒症(CLP)组,采用改良的盲肠结扎穿孔术(CLP)制作大鼠脓毒症模型.采用流式细胞术检测CD14+单核细胞表面人类白细胞抗原-DR基因(HLA-DR)表达率、Treg细胞及TH17细胞比例;酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-6、IL-10、肿瘤坏死因子(TNF)-α、转化生长因子(TGF)-β、白细胞介素(IL)-17炎性因子蛋白表达.结果 与假手术组比较:(1)伴随着脓毒症病情的发展,大鼠出现明显的免疫抑制,CD14+单核细胞HLA-DR表达率＜30％,IL-10/TNF-α比值(27.41 ±7.04比6.63 ±2.60)明显增高(P＜0.01).(2)术后96 h脓毒症大鼠Treg细胞[(11.91±3.88)％比(6.57±2.60)％,P＜0.01]和Th17细胞[(5.14±0.29)％比(2.85±0.07)％,P＜0.01]表达明显增高.(3)术后96 h脓毒症组前炎性细胞因子IL-6[(42.31±15.89) ng/L比(6.32 ±3.18) ng/L,P＜0.01]、IL-10[(69.89 ±20.78) ng/L比(13.58±5.37) ng/L,P＜0.01]、TNF-α[(5.03±3.10) ng/L比(2.77±1.10) ng/L,P＜0.01]、TGF-β[(4.99±2.01) ng/L比(1.88±1.07) ng/L,P＜0.01]、IL-17[(92.77±11.64) ng/L比(7.58±2.30) ng/L,P＜0.01]表达明显增高.结论 伴随着脓毒症病情的发展,大鼠出现明显的免疫抑制;在大鼠脓毒症的发生发展中,Treg细胞介导的免疫抑制及Th17细胞介导免疫激活反应同时存在;脓毒症细胞因子微环境变化可能是导致Treg细胞/Th17细胞失衡的原因之一.     

兔肺VX2肿瘤手术切除模型中外周血CD4+CD25+调节性T细胞的变化和意义

目的 观察外周血CD4+CD25+调节性T细胞水平在兔肺VX2肿瘤手术切除模型中的变化.方法 将30只纯种新西兰白兔随机分成3组.A组:建立兔肺VX2肿瘤手术切除模型;B组:建立兔肺VX2肿瘤模型,不进行肿瘤切除术;C组:不建立兔肺VX2肿瘤模型,仅行兔肺叶切除术.3组实验兔在相同的时间点抽取外周血,经流式细胞仪分析测定外周血CD4+CD25+调节性T细胞的水平.结果 兔在种植肿瘤后外周血CD4+CD25+调节性T细胞的水平增高(P＜0.01);手术切除兔肺VX2肿瘤后15 d和30 d,实验兔外周血CD4+CD25+调节性T细胞水平较术前下降(P＜0.05),而不进行肿瘤切除的实验兔外周血CD4+CD25+调节性T细胞水平持续增高(P＜O.05).结论 外周血CD4+CD25+调节性T细胞水平升高可能与肿瘤的发生发展相关;手术切除肿瘤可减少肿瘤负荷,有利于机体抗肿瘤免疫功能的恢复.

Abstract：

Objective To analyze the change of CD4+ CD25 + regulatory T cells in peripheral blood in rabbit' s lung VX2 tumor resection model. Methods Thirty New Zealand rabbits were divided into 3 groups: group A, 10 rabbits with lung VX2 tumor receiving lobectomy; group B, 10 rabbits with lung VX2 tumor receiving no operation; group C, 10 rabbits without lung VX2 tumor receiving lobectomy only.The peripheral blood of rabbits was drawn. All the samples were analyzed by flow cytometry (FCM). Results The proportion of the CD4 + CD25 + regulatory T cells in peripheral blood of the rabbits was increased after the implantion of VX2 tumor ( P ＜ 0. 01 ), that in the rabbits with VX2 lung tumor receiving tumorectomy was obviously decreased at the 15th and 30th day after the operation (P ＜ 0. 05 ), but that in the rabbits with VX2 lung tumor receiving no operation did not have the same change ( P ＜ 0. 05 ). Conclusion The increase of the proportion of CD4 + CD25 + regulatory T cells in peripheral blood is related to immunosuppression. The functional disorder of immune system recovers in a certain extent after resection of the tumor.     

大鼠CD4^＋CD25^＋调节性T细胞的分离及功能鉴定

目的：研究利用免疫磁珠分选法稳定分离正常大鼠脾脏CD4^＋CD25^＋调节性T细胞的方法。方法：采用免疫磁珠两步法分离大鼠脾组织CD4^＋CD25^＋T细胞。首先采用藻红蛋白（PE）标记的抗CD25抗体和抗PE多功能磁珠试剂盒阳性分选CD25^＋T细胞,再用抗异硫氰酸荧光素（FITC）标记抗体和抗IgG磁珠阳性分选获得CD4^＋CD25^＋T细胞。分离后的细胞经流式细胞仪检测分离纯度,台盼蓝染色检测细胞存活率,体外增殖实验检测其对CD4^＋CD25^-T细胞的免疫抑制作用。结果：两次阳性分选后获得的CD4^＋CD25^＋T细胞纯度为（90.4±1.6）%,细胞存活率为（92.6±2.4）%。体外增殖实验表明,CD4^＋CD25^＋T细胞能明显抑制CD4^＋CD25^-T细胞的增殖（P〈0.01）。结论：采用免疫磁珠法两次阳性分选,可稳定地获得纯度理想并有免疫抑制功能的大鼠CD4^＋CD25^＋T细胞。     

Presence of Functional Mouse Regulatory CD4+CD25+T Cells in Xenogeneic Neonatal Porcine Thymus-Grafted Athymic Mice

Xenotransplantation with porcine thymus is emerging as a possible means to reconstitute host cellular immunity and to induce immune tolerance in rodents and large animals. However, the presence of regulatory T cells (Treg cells) in this model needs to be determined. We herein demonstrated that efficient repopulation of mouse CD4+CD25+Treg cells was achieved in Balb/c nude mice by grafting neonatal porcine thymic tissue (NP THY). Mouse CD4+CD25+T cells expressed normal levels of Foxp3 in NP THY-grafted nude mice. Furthermore, these CD4+CD25+Treg cells showed significant inhibitory effects on the cell proliferation or interleukin-2 products of syngeneic T cells to alloantigens, Con A or a peptide antigen, although the potent immunosuppressive function might be lower than CD4+CD25+Treg cells in Balb/c mice. CD4+CD25+T cells in NP THY-grafted nude mice showed significantly stronger inhibition on the response to donor porcine antigens of CD4+CD25(-)T cells than CD4+CD25+Treg cells in Balb/c mice. Both CD4+CD25+Treg cells in NP THY-grafted nude and Balb/c mice prevented the development of autoimmune disease mediated by syngeneic CD4+CD25(-)T cells in a similar efficient way in the secondary recipients. These findings provide evidence for the potential involvement of CD4+CD25+Treg cells in keeping self-tolerance and transplant tolerance in this xeno-thymus transplantation model.     

The significantly enhanced frequency of functional CD4CD25Foxp3 T regulatory cells in therapeutic dose aspirin-treated mice

CD4⁺CD25⁺Foxp3⁺ regulatory T (Treg) cells, produced in the thymus or periphery as a functionally mature T cell subpopulation, play pivotal roles in maintenance of self-tolerance and negative regulation of immune responses. Aspirin (ASA) is widely used to reduce pain, the risk of cardiovascular diseases and allo-graft rejection. However, the effect of ASA on CD4⁺CD25⁺Foxp3⁺ Treg cells has yet to be determined. The frequency, phenotype and immunosuppressive function of CD4⁺CD25⁺Foxp3⁺ Treg cells were detected in BALB/c mice treated with low or high doses of ASA for 4 weeks. ASA significantly decreased the percentage and number of CD4⁺ T cells in the periphery, while ASA remarkably increased the percentage of CD4⁺CD25⁺Foxp3⁺ Treg cells in CD4⁺T cells. The total cell numbers of thymocytes were significantly decreased in ASA-treated mice, but the number of CD4⁺ CD25⁺Fxop3⁺ cells and its ratio in CD4⁺CD8⁻ thymocytes were markedly enhanced in the thymi of ASA-treated mice. The phenotype of CD4⁺CD25⁺ Treg cells, including the expressions of CD44, CD45RB, CD62L, CD69, GITR and CTLA-4, did not show detectable changes in ASA-treated mice. CD4⁺CD25⁺ Treg cells in ASA-treated mice exhibited unimpaired immunosuppressive function on CD4⁺CD25⁻ T effector cells. ASA significantly enhanced the frequency of functional CD4⁺CD25⁺Foxp3⁺ Treg cells in mice in a therapeutic dose range. The different effects of ASA on CD4⁺CD25⁺Foxp3⁺ Treg cells and CD4⁺CD25⁻ T cells may potentially make hosts susceptible to tolerance induction which would be beneficial for tolerance induction in patients with autoimmune diseases or allo-grafts. This study may have potential impacts in the clinical application of ASA.     

阻断ICOS/B7h信号的供体特异性输血对移植受体CD4+CD25+调节性T细胞的影响

目的 观察阻断ICOS/B7h信号的供体特异性输血(DST)对异基因小鼠心脏移植术后体内CD4+CD25+调节性T细胞(Treg)的影响.方法 按陈氏方法建立小鼠颈部异位心脏移植模型,实验分3组,异基因组及同基因组:供心分别来源于BALB/C和C57BL/6小鼠,受体均为C57BL/6小鼠,未予治疗.治疗组:移植当天给予受体鼠(C57BL/6)尾静脉注射5×106 ICOS-Fc靶定的供体(BALB/C)脾B淋巴细胞,d0～6连续给予受体鼠尾静脉注射ICOS-Fc 200 μg/d.术后统计各组移植物的存活时间,通过流式细胞术检测受体鼠外周血中CD4+CD25+Treg的亚群比例,利用逆转录-聚合酶链反应(RT-PCR)检测移植物中FOXP3的mRNA表达,在混合淋巴细胞反应中检测CD4+CD25+Treg对CD4+CD25-效应T细胞(Teff)的增殖抑制效率.结果 与异基因组比较,治疗组心脏移植物存活时间明显延长[(84.38±29.14)d比(7.00±0.76)d,P＜0.01].各组中,治疗组受体外周血中CD4+CD25+Treg亚群比例显著上调[(15.60±5.69)%,P＜0.01].与其他两组比较,治疗组心脏移植物中FOXP3 mRNA表达显著上调.以正常鼠为对照,耐受鼠脾脏中获取的CD4+CD25+Treg能够更高效地抑制CD4+CD25-Teff在混合淋巴细胞培养中的增殖效应.结论 通过阻断ICOS/B7h信号的DST可以诱导异基因小鼠心脏移植耐受,CD4+CD25+Treg在耐受的形成与维持中均起着重要作用.     

树突状细胞和CD4+CD25+调节性T细胞在免疫耐受中的作用及关系

诱导免疫耐受具有高效、低毒、经济等优点,在器官移植中有着极其重要的意义.在建立免疫耐受中,树突状细胞(DC)起到克隆清除、克隆无能、表达T细胞抑制因子、选择性激活辅助T细胞和诱导调节性T细胞尤其是CD4+CD25+调节性T细胞的产生等作用.而CD4+CD25+调节性T细胞主要以DC为作用靶点,影响其分化成熟,抑制DC向...     

类风湿性关节炎患者外周血环瓜氨酸肽抗体与CD4+CD25+调节性T细胞检测的意义

目的 探讨类风湿性关节炎(RA)患者外周血环瓜氨酸肽(CCP)抗体、CD4+ CD25+调节性T细胞检测的意义.方法 54名符合1987年美国风湿病协会(ACR)修订的RA诊断标准的患者为实验组,20名正常体检健康人员做为对照组.用酶联免疫吸附试验(EUSA)检测血清CCP抗体,用流式细胞仪测定外周血CD4+ CD25+调节性T细胞水平,RA组患者均行双手和双足X线检查评价骨关节侵蚀情况.结果 RA组CCP抗体的阳性率为66.7％,较对照组明显增高(P＜0.01);RA组及对照组CD4+ CD25+调节性T细胞表达水平分别为(3.41±1.83)％、(4.23±2.16)％,两组差异有统计学意义(P＜0.05);RA患者CCP抗体阳性组与阴性组CD4+ CD25+调节性T细胞表达水平分别为(3.17±1.57)％、(3.59±1.84)％,差异无统计学意义(P＞0.05),两组骨关节侵蚀发生率分别为72.2％、16.7％,差异有统计学意义(P＜0.05),CCP抗体阳性组骨关节侵蚀率与CD4+ CD25+调节性细胞水平无明显相关(r=-0.126,P＞0.05).结论 CCP抗体可以作为RA的重要实验室检测指标;CCP抗体阳性RA患者较阴性患者更易发生骨关节侵蚀,但骨关节侵蚀的发生可能与CD4+ CD25+调节性T细胞的表达水平降低无关.