老年慢性阻塞性肺疾病患者外周血T细胞表达趋化因子受体3的研究

目的 探讨趋化因子受体3(CXCR3)在老年慢性阻塞性肺疾病(COPD)发病中的作用.方法 研究对象均为65岁以上的老年人,分为COPD急性加重期组(20例)、COPD稳定期组(17例)及对照组(14例),分离外周血单个核细胞,流式细胞仪检测CD4+和CD8+T细胞分别占总T细胞的百分比,CD4+CXCR3+T细胞占总CD4+T细胞的百分比,CD8+CXCR3+T细胞占总CD8+T细胞的百分比,以及CD4+CXCR3+和CD8+CXCR3+T细胞上CXCR3的平均荧光强度(MFI).结果 ①三组间及COPD急性加重期组治疗前后CD4+、CD8+T细胞分别占总T细胞的百分比,差异无统计学意义.②CD8+CXCR3+T细胞占总CD8+T细胞的百分比及CD4+CXCR3+T细胞占总CD4+T细胞百分比:COPD急性加重期组较COPD稳定组及对照组明显降低,COPD稳定组较对照组明显升高,差异有统计学意义,P＜0.05.CD4+CXCR3+和CD8+CXCR3+T细胞上CXCR3的MFI三组间差异无统计学意义.③COPD急性加重期组经治疗后CD8+CXCR3+T细胞占总CD8+T细胞的百分比及CD4+CXCR3+T细胞占总CD4+T细胞百分比,CD4+CXCR3+和CD8+CXCR3+T细胞上CXCR3的MFI,均明显降低,差异有统计学意义,P＜0.05.结论 COPD稳定期T细胞上CXCR3的过度表达在COPD慢性炎症过程中起作用.     

吸烟对慢性阻塞性肺疾病患者稳定期支气管肺泡灌洗液中T细胞功能影响的临床研究

目的 评价吸烟对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者稳定期支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中T细胞功能的影响.方法 流式细胞术测定吸烟的COPD患者稳定期BALF中CD3+、CD4+、CD8+T细胞和CD4+/CD8+水平.结果 与非COPD组相比,COPD患者稳定期BALF中CD3+、CD4+、CD4+/CD8+水平明显下降(P＜0.05),CD8+明显升高(P＜0.05);在COPD患者中,稳定期BALF中CD3+、CD4+、CD8+T细胞无明显差异(P＞0.05),但持续吸烟组CD4+/CD8+明显下降(P＜0.05).结论 COPD患者稳定期气道T细胞功能明显异常,其中持续吸烟的COPD患者气道CD4+/CD8+下降最为显著.     

趋化因子受体3在老年COPD中的作用

目的探讨老年慢性阻塞性肺疾病(COPD)发病机制中趋化因子受体3(CXCR3)的作用。方法选择COPD急性加重期、稳定期患者和对照组各30例,分别作为A、B、C组,用流式细胞术进行检测。结果CD+4CXCR3+T细胞占总CD+4T细胞的百分比及CD+8CXCR3+T细胞占总CD+8T细胞的百分比的比较中,A组治疗前均明显低于B、C组(P均0.05)治疗后CD+4CXCR3+T细胞占总CD+4T细胞的百分比、CD+8CXCR3+T细胞占总CD+8T细胞的百分比、CD+4CXCR3+T与CD+8CXCR3+T细胞MFI相比治疗前均显著下降(P0.05)。结论 COPD稳定期T细胞上CXCR3+表达明显增多,在COPD慢性炎症反应中可能发挥着重要作用。     

CD8+T细胞在慢性阻塞性肺疾病发病机制中的作用

研究表明CD8^＋T细胞在吸烟导致的慢性阻塞性肺疾病（COPD）发病的各个环都起着非常重要的作用。本文就CD8^＋T细胞在COPD发病机制中的研究进展作一简要综述。     

趋化因子受体 3 在老年 COPD 中的作用简

目的 探讨老年慢性阻塞性肺疾病（COPD）发病机制中趋化因子受体3 （CXCR3）的作用.方法 选择COPD急性加重期、稳定期患者和对照组各30例,分别作为A、B、C组,用流式细胞术进行检测.结果 CD＋4CXCR3＋T细胞占总CD＋4T细胞的百分比及CD＋8CXCR3＋T细胞占总CD＋8T细胞的百分比的比较中,A组治疗前均明显低于B、C组（P均〈0.05）治疗后CD＋4CXCR3＋T细胞占总CD＋4T细胞的百分比、CD＋8CXCR3＋T细胞占总CD＋8T细胞的百分比、CD＋4CXCR3＋T与CD＋8CXCR3＋T细胞MFI相比治疗前均显著下降（P〈0.05）.结论 COPD稳定期T细胞上CXCR3＋表达明显增多,在COPD慢性炎症反应中可能发挥着重要作用.     

特异性和非特异性免疫细胞在COPD发病机制中的作用

COPD是一种暴露于有害气体或颗粒而导致的持续性气流受限,呈进行性发展,不完全可逆的慢性气道及肺部炎症.COPD是许多炎症通路介导的复杂病理过程,是成年人发病及病死的主要原因之一.巨噬细胞、中性粒细胞、树突状细胞、CD8+T和CD4+T细胞亚群是COPD发病过程中的主要炎症细胞,这些炎症细胞与人体疾病研究还不清楚,是如何参与宿主防御,免疫调节和自身免疫作用有待进一步研究.本文旨在对各种炎症细胞及在COPD免疫机制中的作用作一综述.     

Study on mRNA expression of CXCR3 on peripheral blood mononuclear cells of patients with chronic obstrucitve pulmonary disease

目的 探讨CXC趋化因子受体3(CXCR3)在慢性阻塞性肺疾病(COPD)发病机制中的作用及临床意义.方法 选择COPD急性加重期、稳定期患者和对照组各30例,采用流式细胞术检测外周血单个核细胞中T细胞亚群CD8+T细胞和CD4+T细胞数,实时荧光定量聚合酶链反应检测CXCR3、干扰素-γ(IFN-γ)、IL-4和干扰素-γ诱导蛋白-10( CXCL10) mRNA的表达.结果 流式细胞术检测结果显示,COPD急性加重期患者PMBCs中CD8+T细胞数较COPD稳定期组及对照组明显增高(P＜0.05);且COPD稳定期组较对照组明显升高(P＜0.05).在mRNA水平,与COPD急性加重期组相比,COPD稳定期组和对照组PMBCs中CXCR3、CXCL10和IFN-γ的表达水平均明显降低(P＜0.05).与对照组相比,COPD稳定期组PMBCs中CXCR3、CXCL10和IFN-γmRNA的表达水平显著上调(P＜0.05).三组间IL-4 mRNA的表达比较差异无统计学意义(P＞0.05).结论 CXCR3可通过募集CD8+T细胞及促进IFN-γ和CXCL10等细胞因子的释放而启动COPD急性加重期的炎症级联反应.因此,CXCR3可作为治疗COPD肺部炎症的新目标.     

树突状细胞与慢性阻塞性肺疾病

慢性阻塞性肺疾病（COPD）是一种慢性炎症性疾病,其病灶局部存在中性粒细胞、巨噬细胞和T淋巴细胞（尤其是CD8^＋T细胞）浸润。吸烟是COPD发病的首要诱因,目前已证实T细胞对于吸烟所致COPD的发生发展具有重要作用;树突状细胞（DCs）是已知的功能最强的抗原提呈细胞,可激活初始T细胞,在诱导和调节免疫应答中发挥关键作用。近年来发现DCs也存在于COPD病灶中,且其功能和数量均发生了改变,提示其可能与吸烟所致COPD的发病机制有关。     

慢性阻塞性肺疾病病人外周血T淋巴细胞的检测及意义

目的检测慢性阻塞性肺疾病(COPD)患者体内各类T淋巴细胞的亚群,了解COPD患者的细胞免疫功能状态。方法采用免疫荧光双标记法和流式细胞仪检测COPD患者外周血中的T淋巴细胞及其亚群。结果与健康对照组相比较,急性期COPD组患者CD3+T细胞及CD4+T细胞亚群下降,CD8+T亚群细胞升高,导致CD4+/CD8+比值下降并倒置;CD4+CD45RA+亚群、CD4+CD45RA+/CD4+CD45RO+比值下降,CD4+CD45RO+亚群上升;CD8+CD45RA+亚群、CD8+CD45RO+亚群和CD8+CD45RA+/CD8+CD45RO+比值均有所上升;自身免疫性疾病有关的CD4+CD28+亚群下降。结论 COPD患者体内存在细胞免疫功能的紊乱。     

调节性T细胞在慢性阻塞性肺疾病中作用的研究进展

调节性T细胞是近年发现的一种在维持外周免疫耐受和预防自身免疫性疾病中发挥重要作用的CD4+T细胞.慢性阻塞性肺疾病(COPD)是一种吸烟所诱发的由T细胞参与的炎症反应和自身免疫性所引起的疾病,我们综述了近年来调节性T细胞的特征及其在COPD中的作用,以便进一步研究此群细胞的功能,为COPD的防治提供新的思路.     

U94 of human herpesvirus 6 inhibits in vitro angiogenesis and lymphangiogenesis

Human herpesvirus 6 (HHV-6) is a lymphotropic virus, but recent observations showed that also vascular endothelial cells (ECs) are susceptible to infection, both in vivo and in vitro. The observation that lymph nodes are a site of viral persistence suggests that lymphatic ECs (LECs) might be even more relevant for HHV-6 biology than vascular ECs. Here, we provide evidence that HHV-6 can infect LECs in vitro and establish a latent infection. Thus HHV-6 infection induces the loss of angiogenic properties both in LECs and in vascular ECs, as shown by the inability to form capillary-like structures and to seal wound scratches. The antiangiogenic effects observed in infected cells are associated to the expression of HHV-6 U94/rep, a latency-associated gene. In fact, transfection of U94/rep or addition of recombinant U94/REP protein to ECs inhibits the formation of in vitro capillary-like structures, reduces migration of ECs, and blocks angiogenesis, rendering rat aortic rings insensitive to VEGF-induced vasculogenetic activity. The ability of U94/rep to block different angiogenetic steps may lead to approaches in the potential control of the proliferation of blood and lymphatic vessels.     

CD5 positive immunoregulatory B cell subsets

B-chronic lymphocytic leukemia (B-CLL) is a heterogeneous disease often expressed as a clonal expansion of CD5+ B cells. We report the characterization of CD5+ B cells from two unique B-CLL patients. Cells from patient 1 coexpressed CD5 (leu-1), CD19 (Leu-12), CD20 (B1), and HLA-DR; they were CD10 (J5), CD21 (B2), CD22 (Leu-14), CD25 (IL2-R1), PCA-1, surface, and cytoplasmic Ig negative. They suppressed normal peripheral blood lymphocyte (PBL) pokeweed mitogen (PWM) -stimulated immunoglobulin (Ig) synthesis greater than 80%. Cells from patient 2 were CD5 (Leu-1), CD19 (Leu-12), CD20 (B1), CD21 (B2), CD22 (Leu-14), HLA-DR, IgM, and kappa positive. They were negative for CD10 (J5), CD25 (IL2-R1), and PCA-1. These cells did not suppress normal PBL PWM-stimulated Ig synthesis but produced a monoclonal IgM kappa protein with rheumatoid factor-like activity. These observations suggest that there are different CD5+ B cell subsets, one immunosuppressive and the other autoreactive.     

壁层上皮细胞的研究

近年来,随着对壁层上皮细胞(parietal epithelial cells,PECs)认识的加深,发现PECs发挥着重吸收蛋白、滤过屏障等重要功能,而且在肾脏受到损伤时,PECs可能作为潜在的干细胞,促进肾脏修复。本文就PECs及相关进展作一综述。     

Transplanted human bone marrow contributes to vascular endothelium

Recent evidence indicates that bone marrow is a source of endothelial progenitor cells that are mobilized into the peripheral blood in response to cytokines or tissue injury. Previously, we showed that functional endothelial cells (ECs) can be clonally derived from phenotypically defined hematopoietic stem cells. To determine the EC potential of human bone marrow and peripheral blood stem cells, blood vessels in sex-mismatched transplant recipients were evaluated. EC outcomes were identified by using a combination of immunohistochemistry and XY interphase FISH. Donor-derived ECs were detected in the skin and gut of transplant recipients with a mean frequency of 2% and could readily be distinguished from CD45-expressing hematopoietic stem cells. None of the >4,000 ECs examined had more than two sex chromosomes, consistent with an absence of cell fusion. Y chromosome signals were not detected in sex-matched female recipients, excluding the vertical transmission of male cells. None of the recipients evaluated before hematopoietic engraftment demonstrated donor-derived ECs, indicating a close linkage between the recovery of hematopoiesis and EC outcomes. Transplantable bone marrow-derived endothelial progenitor cells may represent novel therapeutic targets for hematopoietic and vascular disease.     

巴曲酶对体外培养下内皮祖细胞数量及功能的影响

目的探讨巴曲酶(DF-521)对内皮祖细胞(endothelial progenitor cells,EPCs)数量和功能的影响。方法采用密度梯度离心法从外周血获取单个核细胞,将其接种在细胞培养板中,分为对照组、低剂量干预组(DF-5210.05BU/ml)、中剂量干预组(DF-5210.1BU/ml)和高剂量干预组(DF-5210.2BU/ml)。激光共聚焦显微镜鉴定"FITC标记的荆豆凝集素"和"DiI标记的乙酰化低密度脂蛋白"双染色阳性细胞为正在分化的EPCs,流式细胞仪检测其表面标记并进一步鉴定EPCs。MTT比色法、改良的Boyden小室及黏附能力测定EPCs的增殖、迁移和黏附能力。结果与对照组比较,不同浓度DF-521干预组均随浓度增加明显提高EPCs的数量;中剂量干预组作用24h时对EPCs的数量影响最为显著,较对照组增加近1倍(P<0.05)。DF-521对于EPCs的增殖、迁移、黏附能力也有显著的改善(P<0.05)。结论DF-521可增加体外培养条件下EPCs的数量并改善其功能。     

树突状细胞与细胞融合后诱导T细胞介导的抗肿瘤效应

目的　研究树突状细胞与小鼠肥大细胞瘤细胞P815融合作为肿瘤疫苗免疫小鼠后抑制肿瘤生长的作用及其机理。方法　Balb/C小鼠DC与P815细胞体外融合 ,形成的杂交瘤分 2次免疫接种同基因小鼠皮下 ,然后用P815细胞攻击免疫的小鼠 ,观察肿瘤的生长情况及免疫小鼠脾细胞特异性CTL功能。结果　DC P815融合率为 3 0 %,杂交瘤接种小鼠能产生明显的抗肿瘤效应 ,其拮抗P815细胞攻击的能力明显强于用灭活的死P815细胞与DC混合 ,和单用死亡P815细胞免疫的小鼠及用生理盐水的对照组小鼠。DC P815杂交瘤免疫接种小鼠的脾细胞特异性CTL ,杀伤活性强于其它各组小鼠。结论　DC与肿瘤细胞融合后作为肿瘤疫苗接种可在体内产生明显的抗肿瘤免疫 ,其机理主要是特异性CTL的作用。     

Mesenchymal stem cells efficiently inhibit the proinflammatory properties of 6-sulfo LacNAc dendritic cells

Mesenchymal stem cells emerged as a promising treatment modality for steroid-refractory graft-versus-host disease, which represents a major complication of allogeneic hematopoietic stem cell transplantation. Dendritic cells (DCs) display an extraordinary capacity to induce T-cell responses and play a crucial role in the pathogenesis of graft-versus-host disease. Here, we investigated the impact of mesenchymal stem cells on the proinflammatory capacity of 6-sulfo LacNAc (slan) dendritic cells, representing a major subpopulation of human blood dendritic cells. Mesenchymal stem cells markedly impair maturation of slanDCs and their ability to secrete proinflammatory cytokines, which was dependent on prostaglandin E₂. In contrast, the release of anti-inflammatory IL-10 was improved by mesenchymal stem cells. Furthermore, mesenchymal stem cells efficiently inhibit slanDC-induced proliferation of CD4⁺ and CD8⁺ T cells and polarization of naïve CD4⁺ T lymphocytes into Th1 cells. These results indicate that mesenchymal stem cells significantly impair the high proinflammatory capacity of slanDCs and further substantiate their potential for the treatment of graft-versus-host disease.     

Phenotypic analysis of nylon-wool-adherent suppressor cells that inhibit the effector process of tumor cell lysis by lymphokine-activated killer cells in patients with advanced gastric carcinoma

The causes of down-regulation of cytotoxic immune responses in cancer patients have not been fully evaluated. We previously demonstrated that T-cell-growth-factor-activated peripheral blood lymphocytes (PBL) with the surface phenotype CD8⁺ CD11b^–, from patients with widespread metastasis of gastric carcinoma, inhibited the effector process of lymphokine-activated-killer(LAK)-cell-mediated cytolysis. In this study, we examined suppressor cell activity in freshly prepared PBL from 18 patients with advanced gastric carcinoma, and 10 normal healthy individuals. The suppressor cell activity was assayed by recording whether or not PBL inhibited directly the effector process of LAK cell cytotoxicity. Most of the PBL suspensions from cancer patients showed that they contained a population of cells that can directly inhibit the effector phase of tumor cell lysis of the cytotoxic cells. To analyze further the PBL responsible for the suppression, the cells were passed over a nylon-wool column. Nylon-wool-adherent cells significantly augmented the suppression, while the cells passing through abrogated the suppressive effect. Most nylon-wool-adherent cells from 10 normal healthy controls did not inhibit the cytotoxic reaction. To determine further the suppressor-effector population in nylon-wool-adherent cells, negative-selection studies using CD8-, CD4- or CD11b-coated magnetic beads, and positive-selection studies using CD8- or CD4-coated magnetic beads were performed. Finally the results suggest that the suppressor-effector cells comprise at least two different surface phenotypes: CD8⁺ T and CD8^–CD11b⁺ cells. The possible role of CD4⁺ T cells and HLA-DR⁺ LeuM3⁺ macrophages as suppressor cells was ruled out in nylon-wood-adherent cells. CD8⁺ T and possibly CD8^–CD11b⁺ cells apparently suppressed the efferent limb of the antitumor immunity. The selective immune suppression mediated by these cells may partly be concerned with escape mechanisms of gastric carcinoma from the host immune surveillance system.Abbreviations TCGF T cell growth factor - PBL peripheral blood leucocytes - LAK lymphokine-activated killer - NK natural killer - CTL cytotoxic T lymphocytes     

脐血、外周血内皮祖细胞分化成内皮细胞的实验研究

目的探讨人的脐血、外周血内皮祖细胞（endothelialprogenitorcells,EPCs）体外分离、纯化、诱导扩增和分化为内皮细胞的可行性,并检测其表型和功能。方法新鲜脐血和健康成年人的外周血,使用Ficoll密度梯度离心法得单个核细胞,在M199培养基中体外培养,3d后去除悬浮细胞,继续培养,诱导EPCs增殖和分化。流式细胞仪检测EPCs标志CD34和内皮细胞特异性标志CD31表型,RTPCR检测ecNOS,flk1/KDR基因水平表达,免疫组化验证蛋白水平表达,并进一步通过NO活性的变化检测内皮细胞的功能。结果流式细胞仪检测,外周血单个核细胞（peripheralbloodmononuclearcells,PBMC）刚分离时,CD34阳性表达率为（1.1±0.8）%,培养3d后为（16.9±6.2）%。细胞形态观察发现,刚分离的单个核细胞呈圆形,形态小,3d后有明显集落形成,7d后梭形细胞线样排列,随培养时间增加,细胞形态逐渐变大,呈现出典型铺路石样改变。脐血单个核细胞（umbilicalcordbloodmononuclearcells,CBMC）和PBMC培养10d后,CD31阳性表达率分别为（76±17）%和（82±9）%。RTPCR检测有内皮细胞特异性成分ecNOS,flk1/KDR的表达。免疫组化染色,细胞膜和细胞浆中有弥漫性棕色出现,呈阳性反应,证实了蛋白水平的表达。培养10d的贴壁细胞随着VEGF浓度增加,NO生成增加,具有内皮细胞的功能。结论脐血,外周血EPCs体外分离,纯化,诱导培养后的贴壁细胞表型检测,大部分细胞具有内皮系标志物,并具有产生NO功能。