多色流式细胞术检测人外周血BCG特异性效应型记忆γδT细胞的表型特征

目的检测卡介苗(BCG)刺激后,PPD+正常人外周血中分泌IFN-γ的TCRγδ亚群及其表型特征。方法分离PPD+正常人外周血单个核细胞(PBMCs),用多色流式细胞术检测TCRγδ亚群;BCG刺激PBMCs后检测γδT细胞IFN-γ分泌水平,并分析BCG特异性γδT细胞表型特征。结果依据PBMCs中TCRγδ和δ2的表达,可将γδT细胞分为3个亚群,即δ2high、δ2low和TCRγδ中非δ2T细胞(non-δ2-γδT)细胞。其中,δ2high和δ2lowT细胞几乎全部(>98%)为CD45RO+,而non-δ2-γδT细胞中只有少部分表达CD45RO,其余大部分不表达CD45RO。δ2high和δ2lowT细胞表型类似,主要为CD45RO+CD62L-CD27-,而non-δ2CD45RO+细胞主要为CD62L-CD27+,non-δ2CD45RO-细胞几乎全部为CD62L-CD27-。BCG刺激PBMCs后,可诱导γδT细胞分泌IFN-γ,且主要为δ2high和δ2low细胞,而non-δ2-γδT几乎不产生IFN-γ。进一步分析BCG特异性γδT细胞的表型特征,结果表明,δ2high-CD45RO+IFN-γ+和δ2lowCD45RO+IFN-γ+细胞主要为CD62L-CD27-和CD62L-CD27+,为效应型记忆细胞。结论 BCG刺激PPD+正常人外周血PBMCs后,主要诱导δ2T细胞分泌IFN-γ,且该细胞表现出CD45RO+CD62L-效应型记忆细胞特征,可能在预防结核早期感染中发挥重要作用。     

八色流式细胞术检测人外周血BCG特异性效应型记忆CD4+T细胞的表型特征

目的:检测BCG刺激后,PPD+正常人外周血中细胞因子产生及其亚群.方法:分离PPD+正常人外周血单个核细胞(PBMC),BCG刺激后检测CD4+和CD8+T细胞细胞因子分泌,并用八色流式细胞术分析BCG特异性T细胞亚群.结果:BCG刺激PBMC后,主要是CD4+T细胞分泌Th1细胞因子(IFN-γ、IL-2和TNF-α),而CD8+T细胞几乎不产生细胞因子.进一步分析分泌细胞因子的细胞亚群,主要是CD4+CD45RO+ CD62L(-)CD27(-)和CD4+ CD45RO+ CD62L(-)CD27+分泌细胞因子.结论:BCG刺激PPD+正常人外周血PBMC后,主要诱导CD4+T细胞分泌细胞因子,且该细胞表现出CD4+CD45RO+ CD62L(-)效应型记忆细胞特征,可能在预防结核感染中发挥重要作用.     

人外周血CD4~+ IL-21~+记忆T细胞的特征

目的:探讨人外周血中白细胞介素21(IL-21)的产生细胞及其特征。方法:分离人外周血单个核细胞(PBMC),分为不刺激或anti-CD3(OKT3)、OKT3+anti-CD28、PMA+ionomycin刺激四个组,流式细胞术(FCM)检测产生IL-21的细胞亚群。PMA+ionomycin刺激PBMC、纯化CD4+、CD4+CD45RA-、CD4+CD45RA+细胞、脐带血单个核细胞(CB-MC),FCM分析产生IL-21细胞的表型特征和IL-21与Th1、Th2、Th17和Th22细胞因子之间的关系。结果:与OKT3、OKT3+anti-CD28相比,PMA+ionomycin能诱导最高量的IL-21产生。产生IL-21的主要细胞为CD4+T细胞,少数CD8+T细胞。CD4+IL-21+T细胞表达CD45RO,不表达CD45RA,其中部分细胞表达CCR6、CCR7或CXCR5。CD4+CD45RA-细胞表达IL-21远高于CD4+CD45RA+细胞。进一步研究表明,PBMC产生IL-21,而CBMC不产生。此外,大约24%的CD4+IL-21+细胞表达IFN-γ,小于10%CD4+IL-21+细胞表达IL-4、IL-17或IL-22。结论:人PBMC在多克隆刺激的条件下,可以诱导IL-21的产生。产生IL-21的主要细胞亚群具有记忆CD4+T细胞的表型。其中一部分CD4+IL-21+T细胞的表型独立于Th1、Th2、Th17和Th22细胞亚群。     

成人麻疹细胞免疫与CD4+ CD25+调节性T细胞的关系

目的 了解CD4+CD25+调节性T细胞(Treg)与麻疹患者细胞免疫的关系.方法 采集34例成年麻疹患者和27例健康对照的外周血,采用流式细胞术进行CD4+CD25+细胞和FoxP3+细胞的检测.用免疫磁珠从外周血单个核细胞(PBMC)分选出CD4+CD25+T细胞和CD4+CD25-T细胞,分别用抗-CD3单克隆抗体、BCG和NV减毒株进行刺激培养,收集培养上清,用ELISA法检测培养上清中的IFN-γ和IL-10.结果 麻疹患者外周血白细胞、淋巴细胞及CD3+CC4·细胞比例明显降低,而CD4+CD25+细胞与FoxP3+细胞的比例则与对照无显著差别;CD4+CD25+细胞明显抑制抗CD3刺激下CD4+CD25-T细胞产生IFN-γ,且患者和对照没有差别,而IL-10的产生没有这种改变.结论 成人麻疹患者的的免疫抑制与Treg的抑制作用无关. 4+CD25+T细胞和CD4+CD25-T细胞,分别用抗-CD3单克隆抗体、BCG和NV减毒株进行刺激培养,收集培养上清,用ELISA法检测培养上清中的IFN-γ和IL-10.结果 麻疹患者外周血白细胞、淋巴细胞及CD3+CC4+细胞比例明显降低,而CD4+CD2 +细胞与FoxP3+细胞的比例则与对照无显著差别;CD4+CD25+细胞明显抑制抗CD3刺激下CD4+CD25-T细胞产生IFN-γ,且患者和对照没有差别,而IL-10的产生     

人外周血CD3~+CD56~+NKT与Vα24~+iNKT、Vβ11~+iNKT细胞的关系及NKT亚群和细胞因子分泌的研究

目的探讨正常人外周血中CD3+CD56+NKT细胞的频率与CD3+Vα24+iNKT、CD3+Vβ11+iNKT细胞之间的关系、表型特征及细胞因子的表达。方法应用流式细胞术,检测CD3+CD56+NKT细胞频率与CD3+Vα24+iNKT和CD3+Vβ11+iNKT细胞之间的关系、CD4及CD8表面分子及细胞因子的表达。结果正常人外周血CD3+CD56+NKT细胞的频率为6.39%±2.34%;只有1.5%CD3+CD56+NKT细胞表达TCRVα24和1.2%CD3+CD56+NKT细胞表达TCRVβ11,0.7%CD3+CD56+NKT细胞同时表达TCRVα24和TCRVβ11;根据CD4和CD8表面分子的表达,可将CD3+CD56+NKT细胞分为66.9%CD4+、20.4%CD8+和11.6%CD4-CD8-3个亚群;当细胞刺激后,50.2%CD3+CD56+NKT细胞分泌IFN-γ,3.3%分泌IL-4,1.5%同时分泌IFN-γ和IL-4。此外,CD4+NKT、CD8+NKT和CD4-CD8-NKT 3群细胞分泌IFN-γ的频率分别为45.1%、70.3%和55.4%,分泌IL-4的频率为6.5%、7.0%和6.9%,同时分泌IFN-γ和IL-4的频率为2.4%、4.7%和3.1%。结论大多数CD3+CD56+NKT细胞与CD3+TCRVα24+iNKT和CD3+TCRVβ11+iNKT细胞是不同的NKT细胞亚群,CD3+CD56+NKT细胞频率小,能分泌大量的细胞因子,参与机体的免疫反应。     

卡介苗刺激后CD8~+T细胞活化、增殖及其调节

目的:探讨卡介苗(BCG)刺激后,结核菌素试验阳性(PPD+)正常人外周血单个核细胞(PBMCs)中CD8+T细胞的活化、增殖、细胞因子产生及调节性T细胞(Treg)对其调节作用。方法:体外用BCG刺激PPD+正常人PBMCs,检测CD8+T细胞的细胞因子产生、活化和增殖。纯化后获得调节性T细胞(Treg)和CD25-细胞,检测Treg对CD8+T细胞增殖的调节作用。结果:BCG诱导CD8+T细胞表达CD69和CD25等活化分子。在低剂量IL-2存在的条件下,BCG诱导CD8+T细胞发生增殖,且增殖的CD8+T细胞大部分表达Granzyme-B。体外BCG短期刺激PBMCs后,CD8+T细胞几乎不产生IFN-γ、IL-2和TNF-α。此外,调节性T细胞抑制CD8+T细胞增殖。结论:BCG诱导CD8+T细胞活化、增殖和表达颗粒酶,Treg抑制CD8+T细胞增殖。     

内毒素耐受状态下脾脏CD4~+Foxp3~+调节性T细胞的比例变化及意义

以5mg/kg浓度LPS腹腔注射C57BL/6小鼠建立内毒素耐受模型,观察注射72h及8d后脾脏大小、重量及总细胞数;HE染色观察脾脏病理学变化;FACS检测CD4+Foxp3+Treg细胞比例并计算其细胞总数;同时检测CD4+Foxp3+Treg细胞功能相关膜分子CTLA-4的相对表达;Ki-67染色法检测CD4+Foxp3+Treg细胞的增殖情况;PI染色法检测CD4+Foxp3+Treg细胞的凋亡情况;Real-time PCR技术检测脾脏中TGF-β的相对表达。结果发现与对照组相比,LPS注射72h后脾脏大小、重量及细胞总数显著增加(P0.05),且发生病理学改变;脾脏中CD4+Foxp3+Treg细胞比例和数量显著减少(P0.05),CTLA-4表达水平显著降低(P0.05);同时,增殖比例明显减少,凋亡比例增加(P0.05);最后,脾脏中TGF-β的相对表达水平显著减少。LPS注射8d后脾脏大小、重量及细胞总数均与对照组相比无差异(P0.05),且脾脏组织结构未见改变;脾脏中CD4+Foxp3+Treg细胞比例及数量仍减少(P0.05),而其表达CTLA-4水平增加(P0.05),且凋亡和增殖比例无明显差异(P0.05);最后,脾脏组织中TGF-β的相比表达显著增加。结果表明,内毒素耐受状态下,CD4+Foxp3+Treg细胞比例发生明显改变,可能与细胞凋亡和增殖变化有关。     

两种多克隆刺激条件下人诱导性Treg表型的多样性变化

目的:研究人诱导性调节性T细胞(iTreg)细胞表型的多样性变化,并比较PMA/Ionomycin和PHA两种多克隆刺激对人iTreg的诱导的异同。方法:用Ficoll密度梯度离心分离出人PBMC,空白对照组直接检测,其余则在细胞培养液(非刺激对照组)、PMA/Ionomycin溶液(PMA/Ionomycin刺激组)或PHA溶液(PHA刺激组)中培养16小时,以流式细胞仪检测CD4+CD25+FoxP3+CD127-、CD4+CD25+FoxP3+IL-2-、CD4+CD25+FoxP3+IL-10+、CD4+CD25+FoxP3+TGF-β+和CD4+CD25+FoxP3+IFN-γ+的表达。结果:空白对照组显示,约4%的CD4+细胞表达CD4+CD25+FoxP3+CD127-,即nTreg。PBMC在细胞培养液培养16个小时,iTreg的表达无明显变化,但经多克隆刺激后,各不同细胞表型的iTreg的表达明显上升(均P<0.01),表明多克隆刺激可以明显诱导人iTreg的产生。PHA对IL-2-和TGF-β+iTreg的诱导比PMA/Ionomycin强(P<0.01),但仅PMA/Ionomycin可以诱导产生CD4+CD25+FoxP3+IFN-γ+iTreg,而PHA不能诱导。刺激后产生的CD4+FoxP3+IFN-γ+PBL表达与CD25水平呈逆相关。结论:多克隆刺激可以诱导产生人iTreg,PMA/Ionomycin和PHA对人iTreg诱导的机制和程度不一样。多克隆刺激后产生的不同细胞表型反映了人iTreg的多样性变化。     

甲状腺与外周血T淋巴细胞亚群比例差异

目的观察正常甲状腺与外周血T淋巴细胞亚群,包括调节性T细胞(regulatory T cells,Treg)、辅助性T细胞1(T helper 1 cells,Th1 cells)和辅助性T细胞2(T helper 2 cells,Th2 cells)的分布,Treg细胞中干扰素-γ(interferon-γ,IFN-γ)、白介素-4(interleukin-4,IL-4)及其mRNA的表达。方法 20名良性甲状腺肿物患者、3名胸腺手术及3名脾脏切除术患者,术前当天留取外周空腹静脉血,术中分别留取甲状腺、胸腺及脾脏标本,分离单个核细胞,流式细胞术检测标本组织及外周血T淋巴细胞亚群差异。磁珠分离法分离甲状腺及外周血CD25+T细胞,RT-PCR检测Treg中IFN-γ和IL-4 mRNA的表达。NKT细胞受体激动剂α-Galcer刺激Treg 7 d后,RT-PCR检测Treg中IFN-γ、IL-4、CD1d、Vα24及Vβ11 mRNA的表达。结果 1)甲状腺组织Th1、Th2细胞比例低于外周血(P0.001),而Th0、Treg细胞比例高于外周血(P0.001)。2)在甲状腺内,几乎所有的CD4+CD25+Foxp3+细胞可以同时分泌IL-4、IFN-γ,RT-PCR提示其IL-4、IFN-γmRNA表达明显高于外周血,而在外周血中,CD4+CD25+Foxp3+细胞并不表达IL-4或IFN-γ。3)在胸腺和脾脏中,未检测到同时表达IL-4、IFN-γ的CD4+CD25+Foxp3+细胞。4)甲状腺内Treg在α-GalCer刺激前后均不表达NKT细胞的表面标志CD1d、Vα24及Vβ11 m RNA,而IL-4和IFN-γmRNA的表达在刺激前后无明显差异。结论正常甲状腺与外周血T淋巴细胞亚群分布存在差异,甲状腺内Treg特异性的同时表达IL-4及IFN-γ。     

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。     

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.     

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.     

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     

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.