肿瘤细胞分泌物抑制树突状细胞的生成和功能

目的：探讨肿瘤细胞对人单个核细胞源树突状细胞（dendritic cells,DC）的生成和功能的影响。方法：培养外周血单个核细胞源DC，体系中加入人肝癌细胞系H7402，结肠癌细胞系HCT－8及正常人骨髓基质细胞系HFCL的培养上清液，以正常培养体系为对照。应用流式技术检测DC的细胞表面标志及吞噬功能；通过[^3H]－TdR掺入法检测DC激活同种异体T淋巴细胞活化的能力。结果：H7402和HCT－8细胞培养上清液作为条件培养的DC，低表达CD1a抗原、共刺激分子（CD86）及MHC-Ⅱ类分子，与对照及基质细胞组比较有明显差异。DC成熟也受到抑制。此外，DC对葡聚糖（Dextran）的吞噬功能下降，对同种异常T淋巴细胞激活的能力较低。结论：肿瘤细胞分泌的某些物质可抑制DC的生成及其相关功能，这可能是肿瘤细胞逃避机体免疫监视的机制之一。     

肺癌患者外周血树突状细胞的体外扩增及鉴定

目的 探索体外扩增成熟树突状细胞(dendritic cell，DC)的方法，并进一步鉴定Dc的形态及表型，为开展肿瘤临床生物治疗奠定基础。方法 取肺癌患者外周血，经淋巴细胞分离液分离得到DC前体细胞，加入生长因子DCGF诱导DC生成，并加入自体肿瘤相关抗原刺激。收集细胞用透射电镜观察，并用流式细胞仪测定细胞表型。结果 应用该法扩增细胞可以得到大量成熟DC。电镜观察DC具有典型的形态。流式细胞仪细胞表型测定CD80为81．8％，HLA—DR为98．3％，CD86为69．8%，CDla为19．7％，CD14为66．9．1％，CD80和HLA-DR较未成熟DC的表达明显增加。结论 肺癌患者外周血体外培养可成功地获得成熟的DC，进一步为肿瘤临床生物治疗奠定了基础。     

肺癌对树突状细胞抑制作用的研究进展

树突状细胞在肿瘤免疫中起重要作用，该细胞的抑制与肿瘤的发展密切相关，本文就肺癌对树突状细胞抑制作用的研究进展作一综述。     

以树突状细胞为基础的肺癌生物免疫治疗

肺癌对人类健康的威胁日益严重 ,据世界卫生组织(ＷＨＯ)报告 ,1996年全球肺癌新发病人数为 132万 ,同年肺癌导致死亡人数为 98 9万人 ,居各类恶性肿瘤死亡率首位[1] 。 1998年上海市恶性肿瘤流行病学调查资料显示 ,男性和女性肺癌发病率分别达到 5 2 6 / 10万和 18 2 / 10万     

树突状细胞在肺癌免疫治疗中的研究进展

树突状细胞（dendric cells,DC）是人体内功能最强的抗原提呈细胞,可摄取抗原并迁移至邻近的淋巴器官,将抗原信息传递给T细胞和B细胞,引发相应的免疫应答,是机体免疫反应的始动者,在诱导高效而特异的抗肿瘤细胞免疫中起关键作用。近年来,随着分子生物学及免疫学的发展,肺癌的生物治疗研究发展迅速,其中以DC为基础的肺癌免疫治疗成为研究热点。本文就DC的生物学特性、肺癌的免疫缺陷及以DC为基础的肺癌免疫治疗方面的研究现状做一综述。     

树突状细胞在非小细胞肺癌治疗中的研究进展

非小细胞肺癌（non—small cell lung cancer,NSCLC）,包括腺癌、鳞状细胞癌、大细胞肺癌,占据了肺癌的大多数并且受到传统治疗方式的限制,标准治疗方法的缺乏促进了NSCLC新疗法的研究和发展。树突状细胞（dendritic cell,DC）作为专职的抗原提呈细胞（antigen—presenting cell,APC）,是机体免疫应答的主要启动者,在免疫应答的诱导中发挥关键作用,为其用于癌症治疗开辟了新的道路。本文就近几年来有关DC的研究进行回顾,重点阐述NSCLC中DC的研究进展。     

CpG ODN加强树突状细胞疫苗抗Lewis肺癌的研究

目的　探讨CpGODN对树突状细胞 (DC)疫苗抗肿瘤作用的影响。方法　应用ODN182 6作为DC的成熟刺激信号 ,体外控制DC充分成熟 ,以混合或融合的方式将肿瘤抗原负载于DC制备DC疫苗。以特异性杀伤细胞活性和淋巴细胞增殖反应测定疫苗的体外免疫活性 ,并将疫苗经小鼠腹腔注射 ,观察治疗和预防实验肿瘤的生长情况。结果　经ODN 182 6刺激后的DC细胞形态呈成熟状态 ,流式细胞仪分析检测刺激前后DC细胞表面分子CD4 0的表达分别为 11和 2 4 (MFI) ,CD86的表达分别为 33和 75 (MFI) ,刺激后的DC培养上清中IL 12的分泌水平为刺激前的 10倍。刺激后DC融合疫苗组CTL活性、T淋巴细胞增殖活性及体内Lewis肺癌移植瘤的抑瘤率均明显高于未刺激的DC融合组 (P <0 .0 5 )。结论　CpGODN能通过诱导DC成熟 ,增强DC疫苗的抗肿瘤作用 ,有效诱导机体产生特异的抗肿瘤反应。未成熟的DC可通过与肿瘤细胞混合的方式 ,获得较好的抗原捕获效果 ,产生一定的抗肿瘤效应 ,而对于刺激成熟的DC则需通过融合手段负载抗原 ,达到有效的抗肿瘤活性。     

树突状细胞疫苗治疗肺癌血源性转移的实验研究

目的 探索应用树突状细胞肿瘤疫苗治疗肺癌血源性转移的可能性。方法 将Ｌewis肺癌细胞经尾静脉注射给Ｃ５７ＢＬ/６j纯系小鼠建立肺癌血源性转移模型,应用树突状细胞负载合成的抗原多肽ＭＵＴ－１,作为肿瘤疫苗进行免疫治疗。存活的小鼠再用１０倍数量的癌细胞皮下攻击,观察其免疫保护效应。这些小鼠的脾脏Ｔ淋巴细胞经纯化后进行体外抗原致敏,然后测定其特异性细胞毒效应。结果 尾静脉注射Ｌewis肺癌细胞可引起多脏器肿瘤播     

树突状细胞免疫治疗非小细胞肺癌的研究进展

树突状细胞（dendritic cell,DC）作为专职的抗原提呈细胞（antigen-presenting cell,APC）,是机体免疫应答的主要启动者,在免疫应答的诱导中发挥关键作用,诱导DC参与的免疫应答为治疗晚期非小细胞肺癌（non-small cell carcinoma,NSCLC）提供了新的思路。本文就目前DC免疫治疗NSCLC的研究进展进行综述。     

树突状细胞免疫治疗晚期非小细胞肺癌的临床观察

目的探讨热休克诱导的细胞株负载树突状细胞（DC）治疗晚期非小细胞肺癌的安全性和有效性。方法38例均接受过常规治疗的晚期非小细胞肺癌患者分成两组,DC治疗组18例,对照组20例。治疗组患者分离患者外周血单核细胞,体外诱导DC,热休克诱导凋亡的细胞株负载自体DC,将DC行皮内回输,观察患者的安全性和有效性,比较两组的总生存期（OS）。结果没有观察到明显的不良反应,DC免疫治疗安全性高;治疗组的中位生存期为9.2月,对照组的中位生存期7.2月,但两组比较没有统计学意义。结论非小细胞肺癌的DC免疫治疗值得进一步研究。     

肺癌血浆VEGF、血循环癌细胞及树突状细胞相互关系的研究

目的　分析非小细胞肺癌 (NSCLC)患者VEGF血浆含量与癌细胞血道转移及树突状细胞 (DC)成熟的关系。方法　外周血获自正常人、肺部良性疾患以及NSCLC患者。血循环中的癌细胞和DC按已建立的流式细胞仪定量分析技术进行检测。血浆VEGF含量采用ELISA方法测定。结果　 12例正常人及 13例肺部良性疾患的血浆VEGF平均含量分别为 15 .7和 13.2 pg /ml,而 12 3例NSCLC患者的血浆VEGF平均含量为 140 .8pg /ml,显著高于正常人及肺部良性疾患者 (P <0 .0 0 1)。正常人及肺部良性疾患者外周血中癌细胞检测均为阴性 ,而 6 9例NSCLC患者血样中 32例阳性 ,阳性率为 46 .4% ,阳性患者的癌细胞平均含量为 0 .471× 10 6/L。正常人及肺部良性疾患者外周血中DC含量分别为 2 9.0和 2 2 .0× 10 6/L ,而 5 4例NSCLC患者的DC含量为 15 .9× 10 6/L ,显著低于正常人及肺部良性疾患者 (P <0 .0 5 )。若以VEGF≤ 5 6pg/m1作为正常值 ,则VEGF增高的NSCLC患者外周血中癌细胞阳性率及其含量明显增高而DC含量显著降低 ,其中以III、IV期患者最为突出。结论　NSCLC患者血浆VEGF增高具有促使癌细胞血道播散和抑制DC成熟的效应。     

人肺癌组织总RNA转染树突状细胞方法的优化

背景与目的树突状细胞(dendriticcell,DCs)作为体内惟一能活化初始T细胞的抗原递呈细胞,是激发机体特异性免疫反应的关键,以DCs为基础构建的疫苗广泛应用于各种肿瘤免疫治疗中。其中RNA转染DCs构建DCs疫苗的方法,因避免了抗原的筛选和鉴定,所需肿瘤量小等优点,特别适于像肺癌这样的缺乏特异性肿瘤抗原、异质性大、抗原性弱的肿瘤。然而,目前缺乏RNA转染DCs的最佳时间和方法的研究。本研究以人肺癌组织总RNA体外转染DCs,试图摸索转染DCs的最佳条件。方法用免疫组化的方法选择肿瘤组织癌胚抗原CEA、粘蛋白MUC1双阳性的10例肺癌患者,一步法提取肿瘤组织总RNA,采集和分离外周血单核细胞,体外培养DCs和T细胞,将RNA于不同时间和不同方法转染DCs,用流式细胞仪检测CEA和MUC1的表达,混合淋巴细胞反应测定DCs刺激自体T细胞增殖能力。结果未成熟期DCs转染人肺癌总RNA后CEA和MUC1表达量(11.33±2.64和39.68±7.25)明显高于成熟期DCs(5.46±1.63和27.17±4.16),且刺激自身T细胞增殖反应能力更强。电穿孔法转染人肺癌总RNA后DCs表达CEA和MUC1的量(20.53±3.64和65.39±9.33)明显多于脂质体转染(11.33±2.64和39.68±7.25)和直接混合法(0.91±0.27和18.53±3.26),且刺激自身T细胞增殖反应能力更强。结论采用电穿孔法将肺癌总RNA体外转染未成熟DCs可以使DCs表达肿瘤抗原量更高,刺激T细胞增殖的活性更强。     

DC/CIK维持治疗晚期非小细胞肺癌的临床观察

目的 评价树突状细胞联合自体细胞因子诱导杀伤细胞（DC/CIK）维持治疗晚期非小细胞肺癌（NSCLC）的疗效和不良反应。方法 根据治疗情况将2010年1月至2014年1月120例含铂方案化疗4～6个周期后疾病稳定的晚期NSCLC患者分为对照组（n=65）和研究组（n=55）,仅研究组化疗后接受DC/CIK维持治疗。对两组进行生存随访并比较中位无进展生存期（PFS）和总生存期（OS）,同时记录不良反应情况。结果研究组的中位PFS为5.0个月,高于对照组的3.5个月,差异有统计学意义（P＜0.05）;研究组的中位OS为8.5个月,与对照组（8.0个月）的差异无统计学意义（P＞0.05）;两组化疗后获不同疗效亚组中位PFS和OS的差异均无统计学意义（P＞0.05）。常见不良反应为骨髓抑制、发热、乏力和关节酸痛,两组不良反应发生率的差异无统计学意义（P＞0.05）。结论DC/CIK维持治疗可延长化疗后疾病稳定晚期NSCLC患者的进展时间,且不良反应轻,患者可耐受。     

化疗联合DC-CTL治疗不同分期非小细胞肺癌的疗效分析

目的:分析化疗联合树突状细胞诱导的细胞毒性T淋巴细胞(dendritic cells and cytotoxic lymphocytes,DC-CTL)对非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的疗效及安全性.方法:收集我院2013年5月至2015年12月期间住院病理确诊为Ⅰb(具有高危因素)、Ⅱa、Ⅱb、Ⅲa、Ⅲb、Ⅳ期的NSCLC患者114例,其中接受肺癌根治术的患者分为术后化疗联合DC-CTL组20例和术后化疗组32例,未行手术患者分为化疗联合DC-CTL组23例和化疗组39例.回顾分析了114例患者的临床特点、疾病控制率(disease control rate, DCR)、中位无疾病生存期(median-disease free survival,M-DFS)、中位无进展生存期(median-progression free survival,M-PFS)、副作用等资料.随访截止至2016年11月.结果:1年DCR:术后化疗联合DC-CTL组(75.0%)vs术后化疗组(43.8%)(P=0.044).M-DFS:术后化疗联合DC-CTL组(19.5个月)与术后化疗组(11.5个月)相比,差异具有明显统计学意义(P=0.025 5).化疗联合DC-CTL治疗与化疗组相比,DCR及M-PFS不具有统计学差异.DC-CTL治疗的副作用显著低于化疗.结论:术后化疗联合DC-CTL治疗可显著提高DCR及延缓NSCLC患者的疾病进展,且安全性高.     

共培养的树突细胞和CIK细胞对肺癌的体内外抑癌作用

背景与目的:细胞因子诱导的杀伤(cytokine-inducedkiller,CIK)细胞是高效的肿瘤杀伤细胞。树突细胞(dendriticcells,DCs)是体内最强的抗原递呈细胞,并且能够提高效应细胞的抗瘤活性。本实验将DCs和CIK细胞共培养观察DCs对CIK细胞的细胞表型、增殖活性及体内外的抗肺癌作用的影响。方法:从健康人外周血单个核细胞中常规诱导出DCs、CIK细胞后,将DCs和CIK细胞按1∶10比例共培养5天获得DC-CIK细胞。流式细胞仪测DC-CIK细胞表型变化,3H-TdR掺入法测定其体外的细胞毒活性,并用肺腺癌细胞株A549建立裸鼠模型观察DC-CIK体内的抗肿瘤效果。结果:在培养第14天,DC-CIK细胞与单独CIK细胞培养组相比,增殖速率提高[(17.0±1.8)倍vs.(10.9±2.0)倍,P<0.05],CD3 CD56 表达水平明显上调[(36.0±4.2)%vs.(25.7±2.9)%,P<0.05],同时对A549细胞的细胞毒活性明显增强(P<0.05)。裸鼠体内实验表明,接种肺癌细胞51天后DC-CIK组、CIK组的抑瘤率分别为62.9%、41.5%,与对照组相比DC-CIK组及CIK组均抑制裸鼠皮下移植瘤的生长(P<0.01),且DC-CIK组与CIK组抑瘤效应差异有统计学意义(P<0.05)。结论:DCs与CIK细胞共培养可使CIK细胞获得更高的增殖活性和更强的抑癌作用。     

树突状细胞分化成熟障碍在肺癌免疫逃逸中的研究

肺癌患者体内存在广泛的免疫逃逸现象,这一事实已在鼠肿瘤模型及癌症患者得到确认.目前所知肺癌免疫逃逸的机制很多,其中树突状细胞(DC)的分化成熟异常导致的功能障碍是肿瘤免疫逃逸的主要机制之一,肺癌微环境已成为造成DC功能不足的重要因素.     

Neuroblastoma-derived gangliosides inhibit dendritic cell generation and function

Neuroblastoma (NB), a tumor of the sympathetic nervous system, is the most common extracranial solid tumor in children. NB-derived gangliosides inhibit the functional activity of T and natural killer cells, contribute to tumor-induced bone marrow suppression, and cause multiple alterations of hematopoiesis, resulting in pancytopenia. However, the role of gangliosides in the regulation of dendritic cell (DC) generation (dendropoiesis) has not been studied. Using murine and human NB cell lines, we demonstrated that coincubation of murine bone marrow progenitors or human CD34+ progenitor cells with NB cells resulted in a significant inhibition of dendropoiesis in vitro up to 90%. The number of DCs was assessed by FACScan determination of CD83+ or CD11c+ cells coexpressing MHC class II and CD86 molecules. In addition, inhibition of antigen-presenting properties of DCs cultured in the presence of NB cells was observed in allogeneic mixed leukocyte reaction (33,508 +/- 1,613 cpm for control DCs versus 17,428 +/- 152 cpm for NB-treated DCs; P < 0.05). Treatment of NB cells with 10 microM DL-threo-1-phenyl-2-decanolylamine-3-morpholino-1-propanol HCl, an inhibitor of glucosylceramide synthase, markedly abrogated ganglioside synthesis and was accompanied by blockade of NB ability to inhibit dendropoiesis. Furthermore, purified gangliosides added to DC cultures significantly inhibited DC generation. The percentage of CD83+ cells decreased from 51.8 +/- 6.1% in the control group to 12.9 +/- 2.7% in cultures treated with GD2 (P < 0.05). Thus, our results demonstrate that NB-derived gangliosides inhibit the generation of functionally active DCs and may play a role in tumor-induced immunosuppression and subsequent tumor escape from immune recognition and elimination.     

Gene-modified dendritic cells for immunotherapy against cancer

Dendritic cells (DCs) are described as professional antigen-presenting cells because of their superior T-cell stimulatory capacity. For this reason, attention is being focused on using DCs for clinical applications to treat cancer patients. Although preclinical studies are promising, the majority of clinical studies with DCs have not fulfilled the expectations, yet. The field of DC biology has progressed rapidly over the past years, leading to several options for the improvement of vaccination. Among the different parameters to investigate, this review focuses on the efficiency and biological and functional consequences of different gene transfer methods into different subsets of human DCs. Another important consideration for DC-based vaccination is the elucidation of the role of maturation and apoptosis during DC differentiation.     

Lung cancer stem cells: Progress and prospects

Epithelial stem cells are critical for tissue generation during development and for repair following injury. In both gestational and postnatal stages, the highly branched and compartmentalized organization of the lung is maintained by multiple, resident stem/progenitor cell populations that are responsible for the homeostatic maintenance and injury repair of pulmonary epithelium. Though lung epithelial injury in the absence of oncogenic mutation is more commonly expressed as chronic lung disease, lung cancer is the most common form of death worldwide and poses a highly significant risk to human health. Cancer is defined by the cell of origin, responsible for initiating the disease. The Cancer Stem Cell Hypothesis proposes that cancer stem cells, identified by stem-like properties of self-renewal and generation of differentiated progeny, are responsible for propagating growth and spread of the disease. In lung cancer, it is hypothesized that cancer stem cells derive from several possible cell sources. The stem cell-like resistance to injury and proliferative potentials of bronchioalveolar stem cells (BASCs) and alveolar epithelial type II cells (AEC2), as well as cells that express the cancer stem cell marker glycoprotein prominin-1 (CD133) or markers for side populations make them potential reservoirs of lung cancer stem cells. The abnormal activation of pathways that normally regulate embryonic lung development, as well as adult tissue maintenance and injury repair, including the Wnt, Hedgehog (Hh) and Notch pathways, has also been identified in lung tumor cells. It is postulated that therapies for lung cancer that specifically target stem cell signaling pathways utilized by lung cancer stem cells could be beneficial in combating this disease.     

Pattern response of dendritic cells in the tumor microenvironment and breast cancer

Breast cancer (BC) is the most common malignant neoplasm and the cause of death by cancer among women worldwide. Its development, including malignancy grade and patient prognosis, is influenced by various mutations that occur in the tumor cell and by the immune system’s status, which has a direct influence on the tumor microenvironment and, consequently, on interactions with non-tumor cells involved in the immunological response. Among the immune response cells, dendritic cells (DCs) play a key role in the induction and maintenance of anti-tumor responses owing to their unique abilities for antigen cross-presentation and promotion of the activation of specific lymphocytes that target neoplasic cells. However, the tumor microenvironment can polarize DCs, transforming them into immunosuppressive regulatory DCs, a tolerogenic phenotype which limits the activity of effector T cells and supports tumor growth and progression. Various factors and signaling pathways have been implicated in the immunosuppressive functioning of DCs in cancer, and researchers are working on resolving processes that can circumvent tumor escape and developing viable therapeutic interventions to prevent or reverse the expression of immunosuppressive DCs in the tumor microenvironment. A better understanding of the pattern of DC response in patients with BC is fundamental to the development of specific therapeutic approaches to enable DCs to function properly. Various studies examining DCs immunotherapy have demonstrated its great potential for inducing immune responses to specific antigens and thereby reversing immunosuppression and related to clinical response in patients with BC. DC-based immunotherapy research has led to immense scientific advances, both in our understanding of the anti-tumor immune response and for the treatment of these patients.