人外周血和扁桃体中Tfh细胞与其他Th细胞亚群之间的关系

目的比较观察人外周血和扁桃体Tfh细胞的表型以及与Th1、Th17、Th22细胞亚群之间的关系。方法分离正常人PBMC及扁桃体单个核细胞,利用anti-CD3+anti-CD28或PMA+ionomycin刺激后,采用ELISA和流式细胞术(FCM)检测其细胞因子的产生,分析Tfh与Th1、Th17、Th22细胞亚群之间的关系。结果与PBMC中CD4+T细胞不同,扁桃体CD4+T细胞高表达CXCR5和CD45RO,低表达CCR7和CD62L;与PBMC中CD4+T细胞相比,扁桃体CD4+T细胞IL-21和IL-17产生水平较高,IFN-γ产生水平较低,IL-22水平无显著差异;外周血和扁桃体CD4+T细胞中均存在一定比例的IL-21+IL-17+双阳性、IL-21+IL-22+双阳性、IL-21+IFN-γ+双阳性细胞,IL-21单阳性细胞在扁桃体CD4+T细胞中所占比例明显高于外周血;外周血和扁桃体CD4+CXCR5+细胞除表达IL-21外,还表达IL-17、IL-22和IFN-γ。结论扁桃体中存在较多数量的Tfh细胞,大多数Tfh细胞是不同于Th1、Th17和Th22的细胞亚群。     

肝癌患者外周血CD3~+T细胞和CD19~+B细胞中程序性死亡蛋白1(PD-1)及其配体表达水平增加

目的 观察肝细胞肝癌(HCC)患者外周血单个核细胞(PBMC)中程序性死亡蛋白1(PD-1)和配体以及γ干扰素(IFN-γ)水平的变化。方法 采集15例早期HCC患者、13例进展期HCC患者和12例健康人(对照组)的外周血并分离PBMC。流式细胞术检测PBMC中PD-1、PD-L1和PD-L2的表达,ELISA检测血清中IFN-γ的水平,利用Pearson相关系数法和One-way ANOVA分析PD-1与IFN-γ的相关性。结果 进展期HCC患者外周血CD3+T细胞和CD19+B细胞中PD-1、PD-L1和PD-L2的表达水平和血清IFN-γ水平显著高于健康人群和早期HCC患者,HCC患者外周血CD3+T细胞和CD19+B细胞PD-1、PD-L1表达水平与IFN-γ呈正相关。结论 肝癌患者PBMC中PD-1、PD-L1和PD-L2分子表达增加,且PD-1、PD-L1与IFN-γ水平有关联。     

流式细胞术鉴定人外周血滤泡辅助性T细胞

目的:用免疫荧光染色和流式细胞术(FCM)鉴定正常人外周血单个核细胞(PBMC)中滤泡辅助性T(Tfh)细胞, 以便对其功能进行进一步研究.方法:密度梯度离心法分离人PBMC, 采用不同荧光素标记的抗CXCR5、 CD4及CD19抗体进行染色FCM分析, 鉴定出表型为CD4+CXCR5+的T细胞亚群.结果:外周血可以检测到CD4+CXCR5+T细胞亚群, 占CD4+T淋巴细胞百分比为(11.09±0.38)%, MFI值为25.78±0.72, 低于B细胞膜表面CXCR表达水平(MFI值为99.6±8.1).结论:健康人外周血存在一定比例的Tfh细胞, 应用FCM可以成功鉴定Tfh细胞, 为进一步研究其功能提供了良好的技术平台.     

过敏性哮喘患者外周血CXCR5~+CD4~+T细胞的变化及其细胞因子表达研究

本研究旨在探讨过敏性哮喘患者外周血CXCR5~+CD4~+T细胞数量、表型变化及其细胞因子的分布特征。选取25例过敏性哮喘患者、25例非过敏性哮喘患者及27例性别和年龄相匹配的健康对照作为研究对象,流式细胞术检测外周血中CXCR5~+CD4~+T细胞数量及表型,胞内细胞因子染色方法检测细胞因子分布情况。结果显示过敏性哮喘和非过敏性哮喘组外周血CXCR5~+CD4~+T细胞比例均高于健康组[(19.24±5.39)vs(8.24±1.68),P0.01;(17.79±4.01)vs(8.24±1.68),P0.01],而患病两组间无明显差异(P0.05)。患病两组患者外周血CXCR5~+CD4~+T细胞表面PD-1[(15.82±2.71)vs(7.64±1.87),P0.01;(14.28±3.68)vs(7.64±1.87),P0.01]、ICOS[(9.82±2.27)vs(4.98±1.76),P0.01;(10.53±2.08)vs(4.98±1.76),P0.01]表达均高于健康组,而患病两组间差异无统计学意义(P0.05)。患病两组CXCR5~+CD4~+T细胞胞内IL-21的表达均显著高于正常对照组(P0.01)。而CXCR5~+CD4~+T细胞胞内IL-4仅在过敏性哮喘患者中高表达(P0.01)。CXCR5~+CD4~+T细胞胞内IL-17、IFN-γ的表达在三组之间无差异(P0.05)。过敏性哮喘组患者IL-4+CXCR5~+CD4~+T细胞、IL-21+CXCR5~+CD4~+T细胞比例与血清总IgE水平呈正相关(r=0.652,P=0.003;r=0.523,P=0.021)。研究结果表明过敏性哮喘和非过敏性哮喘两种表型哮喘在外周血CXCR5~+CD4~+T细胞数量、表型上未见差异,一群高表达IL-4的CXCR5~+CD4~+T细胞可能在过敏性哮喘疾病机制中具有重要意义。     

SLE患者外周血中CD4~+CXCR5~+T细胞表达ICOS和PD1的相关性研究

本研究检测ICOS、PD-1在SLE患者外周血CD4+CXCR5+T细胞上的表达水平并分析其与SLE临床特征的相关性。采用流式细胞仪检测60例SLE患者及40例健康对照组外周血中CD4+CXCR5+T细胞占CD4+T细胞的比例,同时检测CD4+CXCR5+T细胞上ICOS、PD-1的表达水平,并分析其与SLE疾病活动指数(SLEDAI)、SLE肾脏损害及临床指标之间的相关性。结果显示,SLE患者外周血中CD4+ICOShighT细胞、CD4+ICOShighCXCR5+T细胞、CD4+PD1highCXCR5+T细胞比例显著高于健康对照组(P0.05);CD4+ICOShighCXCR5+T细胞比例与SLE患者的抗dsDNA抗体水平呈正相关(P0.05),与SLEDAI评分、免疫球蛋白、C3、C4、ESR、CRP无相关性(P均0.05);SLE患者血清IL-21水平较健康对照组显著升高(P0.05)。提示,循环外周血中生物标志分子高表达的CD4+CXCR5+T细胞异常,可能参与SLE的发病。     

哮喘患者急性加重期滤泡辅助型T细胞的分化水平及临床意义

目的观察滤泡辅助型T细胞(Tfh细胞)在支气管哮喘患者急性发作期和治疗后的分化水平,探讨Tfh细胞是否在哮喘中存在分化异常及其临床意义。方法采用流式细胞术检测健康志愿者、哮喘患者急性发作期及治疗后外周血单个核细胞中CD4~~+CXCR5~~+T细胞、CD4~~+ICOS~+T细胞、CD4~~+CXCR5~~+ICOS~~+T细胞的比例;反转录PCR法检测CD4~+T细胞中Bcl-6mRNA的水平;ELISA检测血浆白细胞介素21(IL-21)的水平;采用直线回归统计学分析IL-21与患者第一秒用力呼气容积/用力肺活量百分比(FEV1/FVC)及第一秒用力呼气容积/预计值百分比(FEV1/Pre)之间的相关性。结果哮喘患者CD4~~+CXCR5~~+细胞、CD4~~+CXCR5~~+ICOS~~+T细胞比例、Bcl-6 mRNA表达水平、IL-21水平均显著高于正常对照组;治疗后哮喘患者CD4~~+CXCR5~~+细胞、CD4~~+ICOS~~+细胞、CD4~~+CXCR5~~+ICOS~~+T细胞表达比例、Bcl-6 mRNA表达水平、IL-21水平均显著低于急性发作时;哮喘患者IL-21表达水平与患者FEV1/FVC、FEV1/Pre存在负相关。结论 Tfh细胞在哮喘急性期存在分化增强,治疗后Tfh细胞分化减弱,提示Tfh细胞可能参与哮喘的炎症反应,其水平可能反映哮喘患者气道受限程度。     

外周血循环滤泡辅助性T 细胞及IL-21 在银屑病中的表达及意义

目的:探讨外周血循环滤泡辅助性T 细胞(Follicular helper T cells,Tfh)及相关细胞因子IL-21 在银屑病患者中的表达水平及其与疾病活动度的关系。方法:收集38 例银屑病患者及32 例健康对照者,流式细胞术检测外周血循环CD4+CXCR5+ Tfh 和CD4+ CXCR5+ ICOS+ Tfh 细胞比例以及Th17 细胞比例;ELISA 检测血清IL-21 浓度;分析这些指标间及与银屑病疾病活动度评分PASI 间的相关性。结果:与健康对照者相比,银屑病患者外周血循环CD4+ CXCR5+ Tfh 和CD4+ CXCR5+ ICOS+Tfh 细胞比例更高,血清IL-21 浓度和Th17 细胞比例亦显著高于对照组(P<0.05);IL-21 浓度与CD4+ CXCR5+ Tfh 和CD4+CXCR5+ ICOS+ Tfh 细胞比例均显著正相关,而与Th17 细胞比例间无显著相关性(P>0.05);且CD4+ CXCR5+ ICOS+ Tfh 细胞比例和血清IL-21 与银屑病疾病活动度PASI 评分显著正相关,而CD4+ CXCR5+ Tfh 则与之无显著相关性(P>0.05)。结论:银屑病患者外周血循环Tfh 比例、IL-21 浓度上调与银屑病患者疾病活动度密切相关,提示Tfh 可能参与银屑病的发生发展过程,这一效应可能是通过分泌高水平IL-21 实现;IL-21 浓度与Tfh 细胞比例相关,而与Th17 细胞比例无相关性提示银屑病患者IL-21可能主要由Tfh 细胞分泌。     

IL-33增强体外培养的外周血单个核细胞细胞因子的分泌以及杀伤功能

目的探讨白细胞介素33(IL-33)是否能在体外增强外周血单个核细胞(PBMC)分泌细胞因子和杀伤功能。方法常规分离培养健康人PBMC,分别经CD3单克隆抗体(m Ab)/CD28mAb/IL-2、CD3mAb/CD28mAb/IL-2/IL-12、CD3mAb/CD28m Ab/IL-2/IL-12/IL-33三种组合刺激培养72 h,收集细胞。经实时定量PCR(qRT-PCR)检测各组细胞γ干扰素(IFN-γ)、颗粒酶B(granzyme B)mRNA的水平;CCK-8法检测各组细胞对A549人肺腺癌细胞的杀伤活性;采用流式细胞术分析各组细胞IFN-γ、程序性死亡蛋白1(PD-1)的水平。结果各组因子组合刺激的PBMC,形态无明显差异;qRT-PCR检测发现,IL-33刺激组的PBMC中IFN-γ、granzyme B mRNA的水平升高;IL-33刺激组的PBMC对A549的杀伤能力更强;IL-33刺激组的PBMC中CD3+CD8+细胞亚群IFN-γ的水平升高、PD-1的水平降低。结论 IL-33能够在体外增强PBMC的效应功能。     

日本血吸虫感染小鼠的脾脏CD8+PD-1+T细胞的功能特性

目的研究日本血吸虫感染后小鼠脾脏CD8+PD-1+T细胞含量及功能的变化情况。方法使用日本血吸虫感染5～6周的C57BL/6小鼠,分离脾脏,制备石蜡切片,HE染色,观察组织病变情况;分离淋巴细胞并计数,运用流式细胞术检测CD8+PD-1+T细胞的含量;使用细胞表面染色的方法观察CD8+PD-1+T细胞的CD25、CD69、CXCR5、CD40L的表达变化;经PMA和离子霉素的刺激,使用细胞内细胞因子染色的方法检测IFN-γ、IL-10和IL-12的分泌情况。结果日本血吸虫感染后,小鼠体质量下降(P0.05),而脾脏质量和体积均明显上升(P0.01);脾脏中CD8+PD-1+T细胞的比例和绝对值数目增加(P0.01);感染组CD8+PD-1+T细胞高表达CD69(P0.05),而感染组CD8+PD-1+T细胞CXCR5分子表达显著下降(P0.05);经PMA与离子霉素刺激以后,CD8+PD-1+T细胞IFN-γ分泌增加(P0.05)。结论血吸虫感染5～6周小鼠脾脏中CD8+PD-1+T细胞为一群主要的活化细胞群可大量分泌IFN-γ,在血吸虫感染疾病发挥重要的免疫调节作用。     

Th17细胞在Graves’病中的变化及意义

目的:探讨Th17细胞在中国人Graves’病(GD)中的变化及意义。方法:应用电化学发光法测定30例初发GD患者及30例健康体检者血清中促甲状腺激素(TSH)、游离三碘甲腺原氨酸(FT3)、游离甲状腺激素(FT4)、甲状腺球蛋白抗体(TgAb)和甲状腺过氧化酶抗体(TPOAb)的水平;采用流式细胞术检测外周血单个核细胞(PBMC)中CD4+IL-17+T(Th17)细胞的数量;用ELISA法测定初发GD患者和健康对照者PBMC产生IL-17的水平;应用实时定量PCR(qRT-PCR)检测PBMC中ROR-γt、IFN-γ、IL-4 mRNA的表达量;采用免疫组化染色检测甲状腺组织中IL-17+细胞的分布和数量。结果:初发GD组外周血中Th17细胞占CD4+T细胞的比例[(20.59±4.63)%]、PBMC分泌的IL-17水平[(83.22±34.28)pg/ml]以及PBMC中ROR-γt mRNA表达(1.67±0.98)均明显高于正常对照组Th17[(11.77±4.18)%](P<0.01)、IL-17[(19.74±5.99)pg/ml](P<0.01)和ROR-γt mRNA(0.92±0.18)(P<0.05)的水平。与正常对照组比较,GD患者PBMC中IFN-γmRNA的表达(0.31±0.07)、IL-4 mRNA的表达(2.53±0.70)均明显降低(P<0.01)。初发GD组患者PBMC中Th17细胞百分率与ROR-γt mRNA表达呈正相关(r=0.5047,P=0.01);与IFN-γmRNA表达量呈负相关(r=-0.5085,P<0.01);与IL-4 mRNA表达量呈负相关(r=-0.5372,P<0.01)。正常对照组甲状腺组织中未发现IL-17+细胞,GD组甲状腺组织滤泡间质内可见散在的IL-17+细胞。结论:初发GD患者PBMC中Th17细胞数量、PBMC分泌IL-17的水平和ROR-γt mRNA表达明显增高。GD患者甲状腺组织中有IL-17+细胞浸润,提示Th17细胞可能参与了中国人GD的发生。     

Monoclonal antibodies against gastric H+ + K+ ATPase

Monoclonal antibodies were prepared against a purified membrane fraction from hog gastric mucosa containing the H+ + K+ ATPase. On sodium dodecyl sulfate gels the molecular weight of this fraction corresponds to a single band of about 95,000. In contrast, on isoelectric focusing gels three groups of peptides are resolved with isoelectric points of 5.7, 6.2, and 8.5. One of the monoclonal antibodies (HK111) was shown to react selectively with the acidic peptide, whereas another antibody (HK113) reacted with the alkaline peptide, showing that the three peptides were antigenically distinct. Both monoclonal antibodies selectively labeled the parietal cell, and antibody HK111 labeled the tubulovesicles of the resting parietal cell and the microvilli of the secretory canaliculus of the secreting cell. This finding suggests translocation of membrane from the tubulovesicles to the secretory surface on stimulation.     

Lower proportion of CD19+IL-10+ and CD19+CD24+CD27+ but not CD1d+CD5+CD19+CD24+CD27+ IL-10+ B cells in children with autoimmune thyroid diseases

Abstract

Introduction: As it is generally known, regulatory B cells (Bregs) control inflammation and autoimmunity. The significance of Bregs in the population of children with autoimmune thyroid diseases (AITD) still offers plenty of potential to explore. The aim of this study was to estimate the expression of Bregs (phenotype CD19⁺CD24⁺CD27⁺IL-10⁺, CD19⁺IL-10⁺, CD1d⁺CD5⁺CD19⁺IL-10⁺ and CD1d⁺CD5⁺CD19⁺CD24⁺CD27⁺) in a paediatric cohort with AITD and in health controls.

Materials and methods: A total of 100 blood samples were obtained from 53 paediatric patients with Graves’ disease (GD) (N?=?12 newly diagnosed, mean age 12.5?±?3.5 and N?=?17 during methimazole therapy, mean age 12.7?±?4.4), Hashimoto’s thyroiditis (HT) (N?=?10 newly diagnosed, mean age 13.3?±?2.9 and N?=?10 during L-thyroxine therapy, mean age 13.7?±?3.4) and compared with healthy controls (C) (N?=?15, mean age 13.1?±?3.1). The expressions of the immune cell populations were analysed by four-color flow cytometry using a FASC Canto II cytometer (BD Biosciences).

Results: There was a decreasing tendency in the number of lymphocytes B producing IL-10 (B10) cells among all B lymphocytes and more widely, also among all lymphocytes, in each study group, as compared to C. We reported a reduction in IL-10 production in Bregs with the expression of CD19⁺CD24⁺CD27⁺IL-10 and CD1d⁺CD5⁺CD19⁺IL-10⁺ in both untreated and treated AITD.

Conclusions: Our data demonstrate that the reduction in the number of Bregs with CD19⁺CD24⁺CD27⁺IL-10⁺ and CD19⁺IL-10⁺ expression could be responsible for breaking immune tolerance and for AITD development in children.     

Na+ absorption in Aplysia intestine: Na+ fluxes and intracellular Na+ and K+ activities

Microelectrodes were used to measure the potential difference (psi m) across the mucosal membrane of epithelial cells lining the villi of isolated Aplysia californica intestine. In substrate-free NaCl seawater medium psi m was -55.1 +/- 1.2 mV. The cell interior was negative relative to the mucosal bathing solution. Intracellular K+ activity, determined in the absorptive cells with single-barreled liquid ion-exchanger microelectrodes, was 383 +/- 15 mM. Since the calculated K+ equilibrium potential exceeds the membrane potential, K+ is accumulated by the intestinal absorptive cell. Intracellular Na+ activity (aiNa) was also determined in the intestinal cells of Aplysia with single-barreled liquid ion-exchanger microelectrodes and was 17.2 +/- 2.5 mM. aiNa was much less than that predicted by the electrochemical equilibrium value for Na+ across the mucosal membrane. From these data the steady-state transapical Na+ and K+ electrochemical potential differences were calculated. Serosal ouabain abolished net sodium absorption as determined by flux measurements. These results are consistent with the operation of a basolateral Na+ - K+ pump.     

Cytoplasmic Na+-dependent modulation of mitochondrial Ca2+ via electrogenic mitochondrial Na+-Ca2+ exchange

To clarify the role of mitochondrial Na(+)-Ca(2+) exchange (NCX(mito)) in regulating mitochondrial Ca(2+) (Ca(2+)(mito)) concentration at intact and depolarized mitochondrial membrane potential (DeltaPsi(mito)), we measured Ca(2+)(mito) and DeltaPsi(mito) using fluorescence probes Rhod-2 and TMRE, respectively, in the permeabilized rat ventricular cells. Applying 300 nm cytoplasmic Ca(2+) (Ca(2+)(c)) increased Ca(2+)(mito) and this increase was attenuated by cytoplasmic Na(+) (Na(+)(c)) with an IC(50) of 2.4 mm. To the contrary, when DeltaPsi(mito) was depolarized by FCCP, a mitochondrial uncoupler, Na(+)(c) enhanced the Ca(2+)(c)-induced increase in Ca(2+)(mito) with an EC(50) of about 4 mm. This increase was not significantly affected by ruthenium red or cyclosporin A. The inhibition of NCX(mito) by CGP-37157 further increased Ca(2+)(mito) when DeltaPsi(mito) was intact, while it suppressed the Ca(2+)(mito) increase when DeltaPsi(mito) was depolarized, suggesting that DeltaPsi(mito) depolarization changed the exchange mode from forward to reverse. Furthermore, DeltaPsi(mito) depolarization significantly reduced the Ca(2+)(mito) decrease via forward mode, and augmented the Ca(2+)(mito) increase via reverse mode. When the respiratory chain was attenuated, the induction of the reverse mode of NCX(mito) hyperpolarized DeltaPsi(mito), while DeltaPsi(mito) depolarized upon inducing the forward mode of NCX(mito). Both changes in DeltaPsi(mito) were remarkably inhibited by CGP-37157. The above experimental data indicated that NCX(mito) is voltage dependent and electrogenic. This notion was supported theoretically by computer simulation studies with an NCX(mito) model constructed based on present and previous studies, presuming a consecutive and electrogenic Na(+)-Ca(2+) exchange and a depolarization-induced increase in Na(+) flux. It is concluded that Ca(2+)(mito) concentration is dynamically modulated by Na(+)(c) and DeltaPsi(mito) via electrogenic NCX(mito).     

Prognostic Values of CD38+CD101+PD1+CD8+ T Cells in Pancreatic Cancer

Programmed death-1 (PD-1), a key immune checkpoint molecule, has been developed as an oncotherapy target for various carcinomas. However, treatment with anti-PD-1 elicited only a minimal effect in pancreatic ductal adenocarcinoma (PDAC). Subsequent studies revealed the existence of a subset of PD-1⁺ T cells coexpressing CD38 and CD101, representing a fixed dysfunctional subpopulation that are not able to be rescued by anti-PD-1 immunotherapy. However, whether this subpopulation of PD-1 expressing CD8⁺ T cells could be useful in predicting PDAC stage or prognosing survival is unknown. In this study, we used flow cytometry and immunofluorescence assay to analyze the expression of CD38 and CD101 in 183 clinical PDAC samples, including 84 of peripheral blood and 99 of surgical tissues. High coexpression of CD38/CD101 on peripheral PD-1⁺CD8⁺ T cells or tumor-infiltrating lymphocytes (TILs) was found to be most significantly correlated with Tumor/Node/Metastasis (T/N/M) classification and clinical stage, in contrast PD-1⁺CD8⁺ T cells could not correlate with T classification. CD38/CD101 co-repression on TILs also correlated with the poor survival in these PDAC patient samples. Our data suggest that CD38/CD101 might represent a more helpful biomarker than PD-1 alone for diagnosis and prognosis of PDAC.     

Ontogeny of Tumor-associated CD4+CD25+Foxp3+ T-regulatory Cells

ABSTRACT

The critical contribution of CD4+CD25+Foxp3+ T-regulatory cells (Treg) to immune suppression in the tumor microenvironment is well-established. Whereas the mechanisms that drive the generation and accumulation of Treg in tumors have been an active area of study, the information on their origin and population dynamics remains limited. In this review, we discuss the ontogeny of tumor-associated Treg in light of the recently identified lineage markers.     

长春新碱诱导的自噬性凋亡与细胞内游离Ca^2＋浓度的相关性研究

目的 研究长春新碱（ＶＣＲ）诱导的Ｌ－０２细胞自噬性凋亡细胞内游离钙离子浓度（［Ｃａ＾２＋］ｉ）的变化,以及自噬特异性抑制剂３－ｍｅｔｈｙｌａｄｅｎｉｎｅ（３ＭＡ）对此自噬性凋亡和［Ｃａ＾２＋］ｉ的影响。方法 应用已建立的ＶＣＲ诱导的Ｌ－０２细胞自噬性凋亡模型,使用电镜、流式细胞术检测细胞;用Ｆｌｕｏ－３／ＡＭ荧光探针经流式细胞仪测定Ｌ－０２细胞平均［Ｃａ＾２＋］ｉ。结果 电镜及流式细胞术检测证实     

Circulating CD3+ CD4+ CD+ T lymphocytes in multiple sclerosis

Triple-antibody flow cytometry was used to search for distinctive populations of peripheral blood lymphocyte immunophenotypes in multiple sclerosis (MS). Using monoclonal antibodies to the cell surface markers CD3, CD4, and CD8, T cell subsets were quantified on a cohort of 31 MS patients (not treated with corticosteroids for at least 6 months), 30 healthy donors, and 14 patients with other autoimmune diseases (also corticosteroid treatment-free for at least 6 months). Untreated MS patients displayed a significantly greater population of CD3+CD4+CD8+ circulating T cells than healthy donors (P = 0.023). Patients with other autoimmune diseases displayed mean populations of CD3+CD4+CD8+ cells greater than normal donors and less than MS, but not significantly different from either. An additional 45 MS patients who had received corticosteroid therapy within the previous 6 months were phenotyped. Treatment of symptomatic MS with corticosteroids was associated with a smaller population of circulating CD3+CD4+CD8+ cells. Some MS patients have significantly greater numbers of peripheral blood T lymphocytes simultaneously expressing CD3, CD4, and CD8 surface markers than healthy donors and this population of cells may be reduced by corticosteroids treatment. This triple positive phenotype may be a manifestation of a systemic immune abnormality in MS.     

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.