CD4~+CD25~+Foxp3~+调节性T细胞与LSCC发病的相关研究

为研究调节性T细胞在喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)、发展中的变化及其参与疾病进展的作用机制,收集2010～2011年上海市五官科医院收治的50例LSCC患者的肿瘤组织和外周血,应用流式细胞术检测CD4+CD25+Foxp3+Treg细胞及趋化因子受体CCR6的表达变化,Real-time PCR法检测转录因子Foxp3以及细胞因子mRNA的表达量。结果发现:LSCC患者外周血中CD4+CD25+Foxp3+Treg的百分比较正常人显著增加,并与临床分期相关;CD4+CD25+CCR6+Treg Foxp3的表达,以及肿瘤组织Foxp3mRNA的表达皆明显高于对照组,且与临床分期、淋巴结转移相关。同时发现,LSCC患者外周血中TGF-β和IL-10mRNA的检出水平分别高于对照组,但IFN-γ、IL-2、IL-12mRNA的水平低于对照组。提示此类Foxp3+Treg属于一类诱导性T抑制细胞(Foxp3+iTreg),可通过产生IL-10和TGF-β抑制LSCC患者的细胞免疫功能。Foxp3的检测可能对判断LSCC的预后有一定价值。     

肺癌患者调节性T细胞的变化及CpG ODN的干预作用

目的:观察肺癌患者外周血调节性T细胞的变化,并探讨CpG ODN的干预作用。方法:分离肺癌患者和健康志愿者(各30例)的外周血单个核细胞(PBMC),用流式细胞仪检测CD4+CD25+调节性T细胞比例,Real-time PCR检测Foxp3基因的表达,ELISA法检测TGF-β和IFN-γ水平。将30例肺癌患者的PBMC随机分为实验组和安慰剂组,分别给予CpG ODN2006或安慰剂CpG ODN1612干预,比较干预前后上述指标的变化。结果:肺癌患者PBMC中的CD4+CD25+调节性T细胞比例、Foxp3基因的相对表达量、TGF-β水平均高于健康对照组,差异具有显著性,但不同病理分型和分期的肺癌患者亚组间比较无显著性差异。两组的IFN-γ水平比较无差异性。CpG ODN干预后CpG ODN2006治疗组的CD4+CD25+调节性T细胞比例、Foxp3基因的相对表达量及TGF-β水平出现了下降,差异有显著性,IFN-γ水平变化无显著性差异。而CpG ODN1612安慰剂组的上述指标在干预前后则无显著性改变。结论:肺癌患者外周血中的CD4+CD25+Foxp3+调节性T细胞比例、TGF-β水平明显升高。CpG ODN2006干预可下调CD4+CD25+Foxp3+调节性T细胞比例及TGF-β水平。     

CD4~+CD25~+Foxp3~+调节性T细胞在子宫内膜癌患者外周血中的表达及意义

探讨在子宫内膜癌患者外周血中CD4+CD25+Foxp3+调节性T细胞的表达情况及意义。采用流式细胞术检测84例术前子宫内膜癌患者及40例子宫肌瘤患者外周血中CD4+CD25+Foxp3+细胞比例及Foxp3平均荧光强度,采用qRT-PCR检测两组患者外周血中Foxp3的mRNA表达情况,同时采用ELISA检测外周血中TGF-β1和IL-17含量。与子宫肌瘤组比较,子宫内膜癌患者外周血中CD4+CD25+Foxp3+Treg细胞的比例虽略有升高但没有统计学意义(P=0.08),而CD4+CD25+细胞内Foxp3的平均荧光强度明显升高(P<0.001)。子宫内膜癌患者外周血中Foxp3的mRNA表达要明显多于子宫肌瘤组(P<0.001)。子宫内膜癌患者外周血中TGF-β1、IL-17的含量要多于子宫肌瘤组。子宫内膜癌患者外周血中的Foxp3+Treg细胞表达增多,这些细胞可能通过增加细胞因子TGF-β和IL-17的分泌从而调节机体对肿瘤细胞免疫反应的方向,最终促进子宫内膜癌的发生和发展。     

结核病患者外周血中CD4~+ CD25~+调节性T细胞水平的检测

探讨CD4~+CD25~+调节性T细胞及其转录因子Foxp3在结核病发病机制中的作用。研究对象为肺结核患者22例(病例组)以及健康对照者23例(对照组)。采用FACS检测外周血CD4~+CD25~+调节性T细胞的百分率,采用real-time PCR检测外周血单个核细胞Foxp3mRNA的表达以及CD4~+CD25~+调节性T细胞与CD4~+T细胞、CD8~+T细胞、IFN-γ和IL-4的相关性。结核病患者外周血CD4~+CD25~+调节性T细胞占CD4~+T细胞的百分率,病例组(3.38±1.23)%高于对照组(1.97±0.62)%,两组比较差异有统计学意义(P0.05)。血清单个核细胞Foxp3 mRNA相对表达水平为134.54±6.76,高于对照组(40.98±2.34,P0.05)。CD4~+CD25~+调节性T细胞与CD4~+T细胞、CD8~+T细胞以及与IFN-γ和IL-4的表达呈负相关。结核病CD4~+CD25~+调节性T细胞数量增加、特异性转录因子Foxp3 mRNA表达上升,由此引发的免疫抑制效应可能是结核病发生发展的重要原因之一。     

人外周血单个核细胞来源CD4~+CD25~+调节性T细胞的体外扩增培养及功能研究

目的建立人外周血单个核细胞来源的CD4+CD25+调节性T细胞体外扩增培养方法 ,研究体外扩增后细胞功能改变。方法 Ficoll-Paque密度梯度分离健康人外周血单个核细胞,免疫磁珠纯化CD4+T细胞,流式分选CD4+CD25+调节性T细胞。用抗人CD3/CD28单抗和IL-2联合刺激CD4+CD25+调节性T细胞,检测其Foxp3、IL-10和TGF-β表达改变。结果人外周血单个核细胞分离纯化得到纯度98%的CD4+CD25+调节性T细胞,Foxp3表达率为95%;使用IL-2加抗CD3/CD28单抗刺激6周后细胞数量可扩增1 000倍;扩增后细胞Foxp3的表达和IL-10、TGF-β的分泌均显著降低。结论本研究成功建立了高纯度大量CD4+CD25+调节性T细胞纯化和体外扩增方法,研究了CD4+CD25+T细胞体外扩增后功能表型的改变。     

中孕期小鼠CD4~+Foxp~(3+)调节性T细胞的比例变化

本实验通过检测中孕期小鼠全身及局部CD4+Foxp3+调节性T细胞的比例变化,探讨妊娠状态下CD4+Foxp3+调节性T细胞与异基因胎儿抗原刺激之间的相互关系。将成熟B ALB/c雌鼠与BALB/c雄鼠或C57BL/6雄鼠交配作为实验组,选取未孕BALB/c雌鼠为对照组。受孕10~12 d后分离小鼠的外周血、脾脏、淋巴结、胎盘,分别用流式细胞学、免疫组化、RT-PCR法检测相关组织中的CD4+Foxp3+调节性T细胞的变化。结果:(1)流式检测实验组中同基因孕鼠和异基因孕鼠脾脏的CD4+Foxp3+T细胞占CD4+T细胞的比例较对照组明显增高(P<0.01);但是同基因和异基因孕鼠之间这类细胞比例的变化并无明显差异(P>0.05)。同样,在淋巴结和外周血中也取得类似结果;(2)免疫组化S-P法显示对照组和实验组均可在脾脏、髂淋巴结组织中表达Foxp3,但是对照组Foxp3+调节性细胞的数目显著低于实验组(P<0.01),但是同基因和异基因妊娠组间无明显差异(P>0.05);(3)RT-PCR法从同基因孕鼠和异基因孕鼠的胎盘局部检出Foxp3 mR-NA的表达。中孕期小鼠全身相关组织中CD4+Foxp3+调节性T细胞的比例显著增高,其扩增并非由于父系来源的MHC抗原所致,而是由妊娠本身所介导。另外,在胎盘局部有Foxp3 mRNA的高表达,由此推测CD4+Foxp3+调节性T细胞可能在母胎界面的局部发挥作用,抑制针对胎儿的免疫排斥。     

CD4~+Foxp3~+调节性T细胞在恶性血液病中的变化及机制研究

目的:分析CD4+Foxp3+调节性T细胞(CD4+Foxp3+Treg)在恶性血液病患者外周血的比例变化,探讨CD4+Foxp3+Treg参与恶性血液病发病的可能机制。方法:用流式细胞仪检测急性白血病、淋巴瘤患者及健康对照外周血CD4+Foxp3+Treg细胞的比例;然后用小鼠的淋巴瘤细胞EL-4和红白血病瘤细胞FBL3的培养上清液与C57BL/6小鼠脾细胞共同培养72小时,RT-PCR检测Foxp3 mRNA的表达。结果:恶性血液病患者外周血CD4+Foxp3+Treg数量显著高于正常对照(14.9±2.92)%、(5.68±1.21)%,P<0.001。小鼠EL-4和FBL3细胞上清液均能够使小鼠脾细胞Foxp3 mRNA表达水平明显增高。结论:恶性血液病患者CD4+Foxp3+Treg比例增高可能导致抗肿瘤免疫功能低下,此外肿瘤细胞分泌的可溶性物质使Foxp3表达增高,增强了CD4+Foxp3+Treg细胞的抑制功能,使肿瘤易于生长和转移。     

特发性血小板减少性紫癜患者CD4~+ CD25~+ Treg细胞和TGF-β1的相关研究

目的:研究特发性血小板减少性紫癜(ITP)患者外周血CD4+ CD25+调节性T细胞(Treg)及相关细胞因子转化生长因子β1(TGF-β1)的表达水平,探讨二者在ITP发病机制中的作用。方法:流式细胞术(FCM)检测31例ITP患者及25例健康志愿者外周血Treg细胞的数量,ELISA方法检测血清中TGF-β1的含量,并进行相关性分析。结果:ITP患者外周血CD4+ CD25+调节性T细胞数量显著低于正常对照组(P0.05),血清TGF-β1的含量也较正常对照组明显减低(P0.05),差异有统计学意义。但CD4+ CD25+调节性T细胞比例与TGF-β1的含量无相关性(P0.05)。结论:ITP患者CD4+ CD25+调节T细胞数量减少与ITP的细胞免疫失调有关,CD4+ CD25+调节性T细胞和TGF-β1在ITP中作用机制复杂,还需要进一步研究。     

子宫内膜异位症患者外周血CD4~+CD25~+Foxp3~+调节性T细胞的变化

观察子宫内膜异位症患者外周血单个核细胞CD4~+CD25~+Foxp3~+调节性T细胞的数量变化,初步探讨其意义。采用流式细胞术检测20例健康对照者(对照组)及46例子宫内膜异位症患者(疾病组临床r-AFS分期:Ⅰ～Ⅱ期26例,Ⅲ～Ⅳ期20例)外周血单个核细胞(PBMC)中CD4~+CD25~+Foxp3~+调节性T细胞数目,并计算CD4~+CD25~+Foxp3~+调节性T细胞占CD4~+T淋巴细胞的百分率;分析不同分期子宫内膜异位症患者外周血CD4~+CD25~+Foxp3~+调节性T细胞的变化。结果显示,与健康对照组相比,疾病组PBMC中CD4~+CD25~+Foxp3~+Treg占CD4~+T淋巴细胞的百分率及CD4~+CD25~+Foxp3~+Treg绝对数均明显升高(P<0.01,P<0.05);疾病组中,Ⅲ～Ⅳ期的PBMC中CD4~+CD25~+Foxp3~+Treg占CD4~+T淋巴细胞的百分比及CD4~+CD25~+Foxp3~+Treg绝对值较Ⅰ～Ⅱ期均明显升高(P<0.01,P<0.05)。提示子宫内膜异位症患者外周血CD4~+CD25~+Foxp3~+调节性T细胞数目和比例增多,可能存在自身免疫调节功能的紊乱,且与病程发展紧密相关。     

Foxp3转染小鼠CD4+CD25-T细胞抑制NK细胞活性

目的： 通过逆转录病毒载体转染Foxp3基因到小鼠CD4+CD25-T细胞,以研究体外诱导获得的调节性T细胞对NK细胞免疫活性的调节作用及其机制。方法： 携带Foxp3基因的逆转录病毒转染初始CD4+CD25-T细胞,以获得持续性高表达Foxp3的CD4+ T细胞模型。CD4+Foxp3+ T细胞与NK细胞共培养后,用［51Cr］标记的YAC-1细胞检测NK细胞的杀伤毒性。在TGF-β阻断实验中,通过Transwell共培养实验以及向细胞共培养体系加入抗TGF-β抗体,并检测NK细胞杀伤毒性。结果：逆转录病毒转染初始CD4+CD25-T细胞,成功建立了表达Foxp3的CD4+T细胞模型,转染后1周Foxp3阳性表达的T细胞比例为38.0%。CD4+Foxp3+ T细胞在与NK细胞共培养的24 h和48 h后,对NK细胞的细胞毒性杀伤效应的抑制率分别为42.9%和22.7%。在CD4+Foxp3+ T细胞与NK细胞的共培养体系中加入抗TGF-β抗体后,其抑制率分别由原来的42.9%(24 h)、22.7%(48 h)变为3.2%(24 h)、2.1%(48 h)。在Transwell共培养实验中与NK细胞直接接触的Foxp3+CD4+T细胞可以诱导NK细胞免疫抑制,而没有直接接触的Foxp3+CD4+T细胞则不能抑制NK细胞的杀伤作用。结论：强制性表达Foxp3的CD4+CD25-T细胞可以在体外发挥免疫抑制作用,可以抑制NK细胞的细胞毒性杀伤作用。转染 Foxp3的CD4+CD25-T细胞对NK细胞发挥作用依赖于细胞之间的直接接触,与转染后T细胞表面表达TGF-β有关。     

B7H1-Ig诱导Tr1细胞向CD4~+CD25~+Foxp3~+Treg细胞转化的研究

为探讨Ⅰ型调节性T细胞(Tr1)与CD4+CD25+Foxp3+Treg之间的转化和相互关系,以预包被而固相化的B7H1-Ig融合蛋白加抗CD3单抗刺激初始CD4+CD62L+T细胞,分析细胞因子及Foxp3表达水平的变化,检测细胞功能;在B7H1-Ig开始刺激时或诱导细胞分化结束后加入重组人TGF-β,观察其对细胞分化的影响。结果显示,B7H1-Ig激活的CD4+T细胞产生高水平IL-10、IFN-γ和IL-5,极低水平的IL-2和IL-4,不表达Foxp3,通过分泌抑制性细胞因子IL-10发挥免疫抑制功能,证实B7H1-Ig可诱导Tr1细胞的产生。同时发现TGF-β不影响B7H1-Ig刺激的初始CD4+T的分化,却可促进B7H1-Ig诱导的已分化Tr1细胞向CD4+CD25+Foxp3+Treg转化,提示在特定条件下,Tr1细胞可转化的CD4+CD25+Foxp3+Treg。研究结果为将来临床应用CD4+Treg治疗免疫失调性疾病奠定了基础。     

哮喘大鼠淋巴液与血液CD4~+CD25~+Treg及IL-10、TGF-β水平比较和地塞米松干预的影响

目的:观察CD4+CD25+Foxp3+调节性T细胞、IL-10、TGF-β在哮喘大鼠淋巴液与血液中的水平,以及地塞米松干预的影响,探讨其与哮喘发病的关系。方法:建立大鼠哮喘模型,收集激发后0、24、48 h淋巴液、血液中淋巴细胞,采用流式细胞术(FCM)检测淋巴液及血液中CD4+CD25+Foxp3+T细胞百分率;采用ELISA方法检测血浆及淋巴液中IL-10、TGF-β水平。结果:哮喘组淋巴液和血液的CD4+CD25+Foxp3+T细胞百分率水平、血浆IL-10和TGF-β浓度在各时间点均低于对照组和治疗组(P0.05);哮喘组淋巴液中CD4+CD25+Foxp3+T细胞百分率水平、IL-10浓度在不同时间点均高于血液水平(P0.05),而TGF-β浓度则低于血液水平(P0.05);治疗组淋巴液和血液中CD4+CD25+Foxp3+T细胞百分率水平、血浆IL-10和TGF-β浓度与对照组相比无显著性差异(P0.05)。结论:哮喘大鼠淋巴液中存在CD4+CD25+Foxp3+调节性T细胞数量以及功能失调,且淋巴液中CD4+CD25+Foxp3+调节性T细胞水平显著高于其血液水平,地塞米松可能通过影响CD4+CD25+Foxp3+调节性T细胞数量以及功能而发挥治疗作用。     

Steady state migratory RelB+ langerin+ dermal dendritic cells mediate peripheral induction of antigen-specific CD4+ CD25+ Foxp3+ regulatory T cells

Tolerance to self-antigens expressed in peripheral organs is maintained by CD4(+) CD25(+) Foxp3(+) Treg cells, which are generated as a result of thymic selection or peripheral induction. Here, we demonstrate that steady-state migratory DCs from the skin mediated Treg conversion in draining lymph nodes of mice. These DCs displayed a partially mature MHC II(int) CD86(int) CD40(hi) CCR7(+) phenotype, used endogenous TGF-β for conversion and showed nuclear RelB translocation. Deficiency of the alternative NF-κB signaling pathway (RelB/p52) reduced steady-state migration of DCs. These DCs transported and directly presented soluble OVA provided by s.c. implanted osmotic minipumps, as well as cell-associated epidermal OVA in transgenic K5-mOVA mice to CD4(+) OVA-specific TCR-transgenic OT-II T cells. The langerin(+) dermal DC subset, but not epidermal Langerhans cells, mediated conversion of naive OT-II×RAG-1(-/-) T cells into proliferating CD4(+) CD25(+) Foxp3(+) Tregs. Thus, our data suggest that steady-state migratory RelB(+) TGF-β(+) langerin(+) dermal DCs mediate peripheral Treg conversion in response to epidermal antigen in skin-draining lymph nodes.     

肺癌患者CD4+CD25high Foxp3+调节性T细胞的格局变化及意义

目的：研究肺癌患者外周血（PBMC）及肿瘤浸润淋巴细胞（TIL）中CD4^＋ CD25^high Foxp3^＋调节性T细胞（Treg）的比例改变，探讨其在抗肿瘤免疫中的调节作用。方法：分离肺癌患者PBMC及TIL，FACS分析CD4^＋／CD8^＋T细胞的比值及CD4^＋ CD25^highT细胞占CD4^＋T细胞的比例。Real-time PCR检测Treg特异性转录因子Foxp3基因在PBMC及TIL中的表达。结果：肺癌患者PBMC及TIL中CD4^＋／CD8^＋比值降低；而CD4^＋ CD25^high T细胞在CD4^＋T细胞中所占比例升高；Foxp3基因仅在TIL中高表达，而在PBMC中低或不表达，表明肿瘤局部的CD4^＋ CD25^high T细胞主要是CD4^＋ CD25^high Foxp3^＋ Treg。结合临床资料分析显示Treg在肺腺癌比例较高。结论：CD4^＋ CD25^high Foxp3^＋ Treg在肺癌患者肿瘤浸润淋巴细胞中明显升高，可能与其通过细胞与细胞间接触抑制CD8^＋T细胞的杀伤效应，最终发挥免疫抑制效应相关。     

Immune compartmentalization of T cell subsets in chemically-induced breast cancer

In cancer, the phenotype and/or the function of T cells may differ according to their distribution through immune-associated tissues, namely immune compartments. Here, in N-methyl-N-nitrosourea (MNU)-induced mammary carcinomas of rat as a relevant model for human breast tumors, the impact of tumor burden on the T cell subsets populating the tumor microenvironment, the tumor-adjacent and -opposite mammary lymph nodes, and the spleen was assessed. In the tumors, ratio of CD8(+) cytotoxic and CD4(+) helper T cells were not significantly different than other immune compartments. On the other hand, most of these cells were further identified with CD4(+) CD25(hi) or CD4(+) Foxp3(+) , CD8(+) Foxp3(+) regulatory phenotype. The selective presence of Tregs in the mammary tumors but not in neighboring-mammary tissue was also confirmed by the expression of Treg-associated genes. The percentage of CD161(+) NKT cells was also significantly increased especially in the tumors and mammary lymph nodes. In the lymph nodes of tumor-bearing animals, in contrast to the spleen, total amount of CD8(+) cells and CD4(+) cells were increased but both of these compartments harbored high numbers of CD4(+) CD25(hi) Treg cells. TGF-β was determined as the major suppressive cytokine secreted by the immune cells of tumor-bearing animals, in addition, proliferation capacity of the T cells was diminished. Hence, the differential distribution of T cell subsets through the spleen, the mammary lymph nodes and the tumor mass in MNU-induced mammary tumor-bearing animals may contribute to a tumor-associated immunosuppression.     

Aurintricarboxylic acid promotes the conversion of naive CD4+CD25- T cells into Foxp3-expressing regulatory T cells

Naive peripheral CD4(+)CD25(-) T cells can be converted into Foxp3-expressing regulatory T cells under appropriate stimulation conditions. Considering that continuous exposure to antigens is one of the prerequisites for the differentiation and maintenance of Treg cells, we investigated whether preventing activation-induced cell death while providing continuous TCR stimulation could promote the expression of Foxp3 in murine naive CD4(+) T cells. Among the several anti-apoptotic agents tested, aurintricarboxylic acid (ATA) was found to induce the in vitro conversion of naive CD4(+) T cells into Foxp3(+) Treg cells with suppressive activity. Neutralizing studies with an antibody against transforming growth factor (TGF)-β revealed that ATA requires the presence of TGF-β to induce Foxp3 expression in naive CD4(+)CD25(-) T cells. Although ATA itself did not activate the Smad signaling pathway, it down-regulated the extracellular signal-regulated kinase and mammalian target of rapamycin signaling cascade in activated T cells. Lastly, combined exposure to ATA and TGF-β had a synergistic effect on the rate of induction and maintenance of Foxp3 expression. These results indicate that ATA could be exploited to efficiently prepare inducible regulatory T cells in vitro and may aid in more precisely identifying the specific signaling pathways that drive Foxp3 expression in T cells.     

Hydrogen-Rich Saline Ameliorates Allergic Rhinitis by Reversing the Imbalance of Th1/Th2 and Up-Regulation of CD4+CD25+Foxp3+Regulatory T Cells,Interleukin-10, and Membrane-Bound Transforming Growth Factor-β in Guinea Pigs

It is well known that CD4+CD25+Foxp3+Treg cells play an important role in the development of allergic rhinitis (AR); the defect of cell numbers and functions contribute to AR. Hydrogen has been proven effective in alleviating symptoms of AR. We herein aim to verify the protective effects of hydrogen on CD4+CD25+Foxp3+Treg cells in guinea pigs with AR and to explore the effect of hydrogen-rich saline (HRS) on CD4+CD25+Foxp3+Treg cells in animals with AR and investigate the underlying anti-inflammatory mechanism. Eighteen guinea pigs were randomly divided into three groups (control group/AR group/AR-HRS group). The guinea pigs were injected with hydrogen-rich saline (AR-HRS group) for 10 days after sensitization. The control group was injected with an equal volume of normal saline. The number of sneezes, degree of runny nose, and nasal-rubbing movements were scored. Peripheral blood eosinophil count was recorded. The proportions of Th1/Th2 of the peripheral blood and the CD4+CD25+Foxp3+T cells in the CD4+T cells of the spleen and peripheral blood were determined by flow cytometry. The content of interleukin (IL)-10 and transforming growth factor (TGF)-β in the serum was detected by enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expression of Foxp3, IL-10, and TGF-β were determined by Western blot, immunofluorescence, and real-time PCR analysis, respectively. Scores of symptoms, number of eosinophils,and nasal mucosa damage were dramatically reduced after HRS treatment. HRS increased the expression of Foxp3, IL-10, TGF-β, and number of CD4+CD25+Foxp3+Treg cells, which were reduced in AR. HRS also revised the dysregulation of Th1/Th2 balance. Both the number and biological activity of CD4+CD25+Foxp3+Treg cells increased with up-regulation of Th1/Th2 after HRS administration. HRS could play a protective role in attenuating AR through improving the proportion and functions of CD4+CD25+Foxp3+Treg cells.     

Tregs调节小鼠肝癌局部引流淋巴结内免疫效应细胞的功能

目的:探讨肿瘤引流淋巴结(TDLNs)内调节性T细胞(Tregs)对局部免疫效应细胞的调节作用。方法:建立小鼠肝癌TDLNs模型,通过免疫组织化学染色和流式细胞仪检测TDLNs内Foxp3+Tregs和CD4+及CD8+T细胞的数量。实时定量PCR测定Foxp3mRNA表达水平。应用酶联免疫斑点法(ELISPOT)检测TDLNs内CD8+T细胞分泌IFN-γ的功能。结果:TDLNs内Tregs和效应性T细胞均明显扩增,Tregs弥散分布于CD8+T细胞聚居区。TDLNs内Foxp3mRNA表达水平显著高于同一接种肿瘤小鼠腹股沟淋巴结(P0.01)和脾脏(P0.01)。Tregs趋向于在TDLNs内聚集,而非其它外周淋巴结位点。荷瘤小鼠的脾脏Foxp3mRNA表达明显高于注射LPS小鼠脾脏。Tregs抑制TDLNs内已初始化的CD8+T细胞分泌IFN-γ的功能,经anti-CD3刺激激活后,CD8+T细胞分泌IFN-γ的功能可恢复。结论:TDLNs内Tregs通过调控CD8+T细胞功能而发挥重要作用,清除Tregs是发挥特异性肿瘤免疫治疗的关键。