肺癌细胞致敏树突状细胞疫苗的体内外抗肿瘤作用

目的:研究以NCI-H460肺癌细胞致敏的树突状细胞(Dendritic cells,DCs)疫苗的抗肿瘤作用。方法:将IL-18基因转染的NCI-H460细胞及未转染NCI-H460细胞与DC融合作为转染融合DC组及融合DC组疫苗,以NCI-H460细胞RNA冲击的DC为冲击DC组疫苗,以未冲击的DC为DC组疫苗。以MTT法检测4组DC疫苗刺激T细胞增殖的作用,用ELISA法测定DC疫苗上清液中IL-12的含量,用LDH法测定DC疫苗对NCI-H460细胞的杀伤作用。裸鼠皮下接种NCI-H460细胞及DC疫苗,观察成瘤时间及裸鼠存活情况;荷瘤裸鼠瘤内注射DC疫苗,比较肿瘤体积。结果:4组DC疫苗均能刺激T细胞增殖,刺激作用强度为转染融合DC组>融合DC组>冲击DC组>DC组;4组均能分泌IL-12,转染融合DC组IL-12的分泌量高于融合DC组,冲击DC组高于DC组;转染融合、融合、冲击DC和DC组疫苗对NCI-H460细胞的杀伤率分别为79.73%、50.68%、35.81%及4.05%。转染融合组裸鼠移植瘤成瘤时间[(12.82±2.85)d]长于冲击DC组[(8.52±1.97)d](P<0.05)和DC组[(8.33±1.63)d](P<0.01);移植瘤内注射疫苗后的肿瘤体积比较,转染融合组<融合组<冲击DC组相似文献   

树突状细胞疫苗与食管癌免疫治疗

随着树突状细胞(DC)体外扩增技术和肿瘤抗原制备、负载体系等DC疫苗制备技术的发展,与食管癌治疗相关的多种DC疫苗相继出现,包括多肽负载DC疫苗、食管癌细胞-DC融合疫苗、核酸负载DC疫苗、自体肿瘤抗原负载DC疫苗等,其中多肽负载DC疫苗及凋亡的自体肿瘤细胞负载DC疫苗等在临床应用中已取得较好疗效.     

肿瘤细胞DC疫苗研究现状

以肿瘤细胞为基础的树突状细胞(DC)疫苗主要有肿瘤细胞抗原肽DC疫苗、肿瘤细胞溶解物DC疫苗、凋亡的肿瘤细胞DC疫苗和肿瘤细胞与DC融合的杂交疫苗.这些DC疫苗为肿瘤的免疫治疗增添了新内容,并在部分恶性肿瘤的治疗中取得了令人振奋的效果,显示出了DC疫苗在恶性肿瘤等疾病治疗中的巨大前景.     

肿瘤细胞DC疫苗研究现状

以肿瘤细胞为基础的树突状细胞（DC）疫苗主要有：肿瘤细胞抗原肽DC疫苗、肿瘤细胞溶解物DC疫苗、凋亡的肿瘤细胞DC疫苗和肿瘤细胞与DC融合的杂交疫苗,这些DC疫苗为肿瘤的免疫治疗增添了新内容,并在部分恶性肿瘤的治疗中取得了令人振奋的效果。显示出了DCU凤苗在恶性肿瘤等疾病治疗中的巨大前景。     

树突状细胞肿瘤疫苗抗肿瘤研究进展

目的:探讨不同树突状细胞(DC)肿瘤疫苗的抗肿瘤机制及应用价值.方法:应用NCBI的PubMed文献数据库系统,以“树突状细胞、疫苗、肿瘤”为关键词检索2001-01-01-2011-12-31文献1 589篇,纳入标准:1)DC抗肿瘤的机制;2)肿瘤抗原肽负载的DC肿瘤疫苗;3)肿瘤全细胞抗原负载的DC肿瘤疫苗;4)肿瘤细胞来源的基因修饰的DC肿瘤疫苗;5)DC-CIK联合修饰疫苗.根据纳入标准符合分析的共30篇文献纳入分析.结果:DC是目前已知人体内功能最强抗原提呈作用的专职抗原呈递细胞,与肿瘤发展有着密切的关系,以DC为基础制备的不同类型肿瘤疫苗提高了抗肿瘤免疫反应,但存在不同的缺点.结论:DC肿瘤疫苗展示了良好的应用前景,对临床免疫治疗将有着积极的作用,但仍有待于进一步深入研究.     

DC/肿瘤融合细胞疫苗制备规范化研究

树突状细胞（dendritic cells,DC）是目前所发现的功能最强的专职抗原呈递细胞（antigen presenting cell,APC）,在激活静息T细胞产生抗肿瘤反应方面发挥着重要的作用。正因为如此,DC肿瘤疫苗的研究和应用倍受人们的关注,其中一种新型的DC肿瘤疫苗引发了研究者们的极大兴趣——DC／肿瘤融合细胞疫苗,其既表达肿瘤的所有抗原成分,     

树突状细胞疫苗治疗恶性淋巴瘤的进展

 目前针对恶性淋巴瘤（ML）的治疗方案还不是十分有效,因此研发治疗淋巴瘤的新型药物具有重要意义。树突状细胞作为一类抗原递呈细胞,目前已开展了肿瘤抗原冲击树突状细胞（DC）免疫治疗ML的研究。从ML相关的各种抗原类型、DC疫苗的制备及临床试验研究等方面阐述近年来DC疫苗免疫治疗ML的进展,并探讨了DC疫苗的应用前景。     

树突状细胞及其在血液系统肿瘤免疫治疗中的应用

树突状细胞(DC)是体内功能最强的抗原提呈细胞,由DC激活的T细胞免疫在抗肿瘤过程中起着主导作用。关于DC肿瘤疫苗在血液系统肿瘤免疫治疗方面应用的结果已显示出DC疫苗在血液系统肿瘤治疗中有巨大的应用前景。就国内外有关DC的生物学特征和DC疫苗在血液系统肿瘤免疫治疗中的研究进展作一综述。     

肿瘤DC疫苗临床应用研究现状

树突状细胞（DC）是体内功能最强的抗原提呈细胞，由DC激活的T细胞免疫在抗肿瘤过程中起着主导作用。关于DC肿瘤疫苗临床应用的结果显示出DC疫苗在恶性肿瘤治疗中有巨大的应用前景。本文将国内外有关DC疫苗临床应用的最新进展作一综述。     

RNA转染树突状细胞肿瘤疫苗的研究进展

树突状细胞（DC）是体内功能最强的专职抗原递呈细胞,能激发并维持机体的初始免疫反应。通过不同方式将肿瘤抗原信息负载DC制备肿瘤疫苗是肿瘤免疫治疗的一个重要方面。而通过RNA转染DC制备核酸肿瘤疫苗是最近发展起来的新型制备疫苗的方法。现对RNA—DC疫苗的研究进展作一综述。     

肿瘤抗原致敏树突状细胞疫苗治疗晚期恶性肿瘤的临床观察

目的:观察肿瘤抗原致敏人树突状细胞(dendritic cell,DC)疫苗治疗各类晚期恶性肿瘤的不良反应,初步观察其治疗疗效。方法:91例非小细胞肺癌、结直肠癌、恶性黑素瘤、肾癌、乳腺癌等晚期恶性肿瘤患者入组,均符合试验的纳入组及排除标准,签署知情同意书,并报医院伦理委员会审查批准。患者单采外周血单核细胞体外培育成DC,用抗原致敏后制备成DC疫苗回输,每周回输1次,3次定义为1个周期。结果:91例患者共接受96个周期疫苗治疗;不良反应主要表现为寒战、发热、肌肉酸痛、皮肤瘙痒、胸闷及一过性全身无力,大部分为自限性;76例进行疗效评价,无完全缓解(complete remission,CR)及部分缓解(partial remission,PR)病例,治疗后稳定(stable disease,SD)31例,进展(progression disease,PD)45例,临床获益率40.8%;85例患者获得随访资料,患者中位达进展时间(median time to progression,mTTP)为2.6个月,生存时间为0.9~30.6个月,中位生存时间为4.5个月,1年生存率为9.2%。结论:肿瘤抗原致敏人DC疫苗治疗各类晚期恶性肿瘤耐受性良好,亦可见临床获益,疫苗临床应用方法有待进一步探讨。     

IL-23基因修饰的树突细胞疫苗联合β-榄香烯对小鼠胰腺癌生长的影响

背景与目的:树突细胞(dendriticcells,DC)疫苗是目前最具应用潜能的治疗性疫苗,功能性细胞因子能显著增强DC的活性及其诱导的宿主抗肿瘤作用,利用细胞因子基因修饰DC是当今肿瘤免疫治疗中最活跃的领域。本研究探讨β-榄香烯联合白细胞介素-23(interleukin-23,IL-23)修饰的DC疫苗对胰腺癌小鼠的协同抗肿瘤作用。方法:克隆并构建IL-23基因真核双表达载体,转染DC并负载肿瘤抗原后制备成疫苗。将IL-23基因转染的DC疫苗、空质粒转染的DC疫苗、未转染的DC疫苗及对照生理盐水分别注射小鼠,体外观察各组小鼠脾脏T淋巴细胞IFN-γ及IL-4的分泌。体内观察β-榄香烯联合DC疫苗对荷胰腺癌小鼠肿瘤生长的抑制作用及对小鼠存活期的影响。结果:基因测序证实IL-23基因克隆及双表达载体构建成功,转染后DC共刺激分子MHC-Ⅰ和MHC-Ⅱ的表达增强。接种IL-23转染DC疫苗后小鼠的免疫防御能力显著增强,有效地延缓并防御接种肿瘤的发生。DC介导的免疫应答促进了IFN-γ生成型Th1细胞的产生,未转染DC疫苗组和空质粒转染DC疫苗组IL-4分泌量与IL-23转染的DC疫苗组比较差异有显著性(P<0.05);IL-23转染DC疫苗组IFN-γ的分泌与其他各组比较差异有显著性(P<0.01)。β-榄香烯联合IL-23转染DC疫苗组肿瘤生长受到显著抑制,该组小鼠存活期与DC组、NS对照组及β-榄香烯组比较差异有极显著性(P<0.01)。结论:IL-23修饰DC疫苗可强化宿主针对特异肿瘤的Th1及CTL的免疫应答,使宿主不仅产生防御性免疫反应而且增强自动免疫能力。β-榄香烯有一定的协同抗癌作用。     

树突状细胞肿瘤疫苗研究进展

树突状细胞(DC)是具有最强抗原提呈能力的专职性抗原提呈细胞.近年来,以DC为基础的肿瘤疫苗研究已成为肿瘤免疫治疗的热点之一.本文综述各种DC肿瘤疫苗的制备方法和目前临床应用现状.     

Glycan modification of the tumor antigen gp100 targets DC-SIGN to enhance dendritic cell induced antigen presentation to T cells

Dendritic cells (DC) have gained much interest in the field of anticancer vaccine development because of their central function in immune regulation. However, the clinical application of ex vivo cultured DC has significant disadvantages. A vaccine that targets dendritic cells in vivo and enhances antigen presentation would be of great benefit. Because of its DC-restricted expression pattern, and its function as an antigen uptake receptor, DC-SIGN is an interesting candidate target structure for human immature DC. Here, we studied whether modification of the melanoma differentiation antigen gp100 with DC-SIGN-interacting glycans enhances targeting to human DC. A high-mannose form of gp100, as protein or as tumor lysate, not only interacted specifically with DC through DC-SIGN but also resulted in an enhanced antigen presentation to gp100-specific CD4(+) T cells. Our results indicate that glycan modification of tumor antigens to target C-type lectin receptors, such as DC-SIGN, is a new way to develop in vivo targeting DC strategies that simultaneously enhance the induction of tumor-specific T cells.     

Improvement of DC vaccine with ALA-PDT induced immunogenic apoptotic cells for skin squamous cell carcinoma

Dendritic cell (DC) based vaccines have emerged as a promising immunotherapy for cancers. However, most DC vaccines so far have achieved only limited success in cancer treatment. Photodynamic therapy (PDT), an established cancer treatment strategy, can cause immunogenic apoptosis to induce an effective antitumor immune response. In this study, we developed a DC-based cancer vaccine using immunogenic apoptotic tumor cells induced by 5-aminolevulinic acid (ALA) mediated PDT. The maturation of DCs induced by PDT-treated apoptotic cells was evaluated using electron microscopy, FACS, and ELISA. The anti-tumor immunity of ALA-PDT-DC vaccine was tested with a mouse model. We observed the maturations of DCs potentiated by ALA-PDT treated tumor cells, including morphology maturation (enlargement of dendrites and increase of lysosomes), phenotypic maturation (upregulation of surface expression of MHC-II, DC80, and CD86), and functional maturation (enhanced capability to secrete IFN-γ and IL-12, and to induce T cell proliferation). Most interestingly, PDT-induced apoptotic tumor cells are more capable of potentiating maturation of DCs than PDT-treated or freeze/thaw treated necrotic tumor cells. ALA-PDT-DC vaccine mediated by apoptotic cells provided protection against tumors in mice, far stronger than that of DC vaccine obtained from freeze/thaw treated tumor cells. Our results indicate that immunogenic apoptotic tumor cells can be more effective in enhancing a DC-based cancer vaccine, which could improve the clinical application of PDT-DC vaccines.     

Oxygen is a master regulator of the immunogenicity of primary human glioma cells

With recent approval of the first dendritic cell (DC) vaccine for patient use, many other DC vaccine approaches are now being tested in clinical trials. Many of these DC vaccines employ tumor cell lysates (TL) generated from cells cultured in atmospheric oxygen (～20% O?) that greatly exceeds levels found in tumors in situ. In this study, we tested the hypothesis that TLs generated from tumor cells cultured under physiologic oxygen (～5% O?) would be more effective as a source for DC antigens. Gene expression patterns in primary glioma cultures established at 5% O? more closely paralleled patient tumors in situ and known immunogenic antigens were more highly expressed. DCs treated with TLs generated from primary tumor cells maintained in 5% O? took up and presented antigens to CD8 T cells more efficiently. Moreover, CD8 T cells primed in this manner exhibited superior tumoricidal activity against target cells cultured in either atmospheric 20% O? or physiologic 5% O?. Together, these results establish a simple method to greatly improve the effectiveness of DC vaccines in stimulating the production of tumoricidal T cells, with broad implications for many of the DC-based cancer vaccines being developed for clinical application.     

冻融人树突状细胞基因疫苗体外抗肿瘤免疫效应

目的：观察人外周血和人脐血来源的冻融树突状细胞（dendritic cell,DC）基因疫苗的形态、表型及体外诱导的CTL抗肿瘤活性。方法：细胞因子扩增人外周血和脐血DC,分别将EBV-LMP2、HPV16E6基因转染2种来源的冻融DC制备疫苗。动态形态学观察和流式细胞术检测疫苗表面分子表达,体外诱导并测定CTL活性。结果：人外周血和脐血冻融DC疫苗均高表达CD80、CD86和CD83,低表达CD14,高表达CD1a,体外均能诱导高效的CTL活性（P=0.001）,并与新鲜DC疫苗差异无统计学意义,P=0.138。结论：负载肿瘤相关病毒抗原基因的冻融DC疫苗保持了功能成熟DC的形态特征,且能诱导高效的特异性抗肿瘤免疫应答。     

肺癌可溶性抗原联合超抗原修饰致敏ＤＣ诱导抗瘤免疫的研究

目的　观察经肺癌肿瘤可溶性抗原（ＴＳＡ）和超抗原金黄色葡萄球菌肠毒素Ａ（ＳＥＡ）联合修饰致敏树突状细胞（ＤＣ）体外诱导抗肺癌的免疫效应。方法　３ｍｏｌ／Ｌ氯化钾提取法获得人肺癌细胞ＧＬＣ ８２的可溶性抗原;从人外周血单个核细胞（ＰＢＭＣ）中诱导扩增ＤＣ,并用流式细胞仪（ＦＣＭ）检测表型;以肺癌ＴＳＡ和ＳＥＡ联合修饰致敏的ＤＣ、单纯肺癌抗原致敏的ＤＣ和未经抗原修饰的ＤＣ分别与同种异体外周血Ｔ淋巴细胞共同孵育,刺激Ｔ淋巴细胞活化增殖（作为效应细胞分别称为ＴＳＡ-ＳＥＡ-ＤＣＬ、ＴＳＡ-ＤＣＬ、ＤＣＬ）,直接活细胞计数法观察增殖倍数;ＭＴＴ法检测不同ＤＣ∶Ｔ淋巴细胞比例的效应细胞对靶细胞ＧＬＣ-８２的体外杀伤效应。结果　诱导出高表达ＣＤ１ａ、ＣＤ８０、ＨＬＡ-ＤＲ的ＤＣ,光镜下具有典型的ＤＣ特性;联合抗原修饰后的ＤＣ具有较强的免疫刺激活性,少量致敏ＤＣ即可强烈激发Ｔ细胞的增殖;ＴＳＡ-ＳＥＡ-ＤＣＬ对靶细胞ＧＬＣ-８２的杀伤率明显高于ＴＳＡ-ＤＣＬ及ＤＣＬ;联合抗原修饰的ＤＣ以１∶１００与Ｔ淋巴细胞共孵后的杀伤肿瘤细胞效应最强。结论　经肺癌ＴＳＡ和ＳＥＡ联合修饰致敏的ＤＣ可强烈激发同种异体Ｔ淋巴细胞活化增殖;肺癌ＴＳＡ联合超抗原ＳＥＡ诱导的ＤＣ疫苗对肺癌细胞有高效杀伤作用,经肺癌ＴＳＡ与超抗原ＳＥＡ联合修饰ＤＣ的活性明显强于单用肺癌ＴＳＡ。     

Immunization of NSCLC patients with antigen-pulsed immature autologous dendritic cells

Only a handful of NSCLC patients have been included in dendritic cell (DC) vaccine clinical trials. We had previously reported a series of 16 individuals with stages IA-IIIB NSCLC who received autologous DC vaccines matured with dendritic cell/T cell-derived maturation factor (DCTCMF). Here we report the results of a continuation study with similar inclusion criteria, immunization protocol, and analysis, using an immature DC vaccine. Of the 14 participants, 7 had undergone surgical resection (stage I/II), with or without adjuvant therapy, and 7 with unresectable stage III had been treated with chemo-radiation alone. Autologous DCs were pulsed with apoptotic bodies derived from an allogeneic NSCLC cell line that over-expresses Her2/neu, CEA, WT1, Mage2, and survivin. DCs were not exposed to any maturation stimulus. Individuals received two intradermal vaccines (average 8.1x10(7) DC per immunization) 1 month apart. Immune responses were measured by IFN-gamma ELISPOT, comparing relative number of antigen-reactive T-cells from pre-vaccine to timepoints post-immunization. Immunologic responses were seen in 4/7 stage III unresectable, and 6/7 stage I/II surgically resected patients, including 3/3 resected patients who had also received adjuvant chemo-radiation. There were no related adverse events. One of seven surgically resected patients recurred and 4/7 stage III patients progressed. Three of five patients with progressive disease showed no immunologic response. Data indicate that immature DC pulsed with apoptotic tumour cells have similar biologic activity to a DCTCMF-matured DC preparation delivered in a similar clinical protocol. Therapeutic efficacy is unknown and clinical outcomes are anecdotal.