顺铂预处理化疗通过免疫调节增强CIK细胞对B16恶性黑色素瘤的抑制作用

目的 观察顺铂( Cisplatin,DDP)预处理化疗联合CIK细胞对B16恶性黑色素瘤的抑制作用,探讨DDP预处理化疗增强细胞因子诱导的杀伤细胞(cytokine - induced killer cells,CIK cells)抑瘤作用的潜在机制.方法 建立C57BL/6小鼠B16黑色素瘤模型,测量肿瘤体积,绘制...     

记忆性干细胞样T细胞在肿瘤治疗应用中的研究进展

记忆性干细胞样T细胞(stem memory T cells,TSCM)是最近几年发现的一个细胞亚群,约占外周血中总的CD4+T细胞和CD8+T细胞的2％～4％,是记忆性T细胞的早期分化阶段,具有记忆性细胞和干细胞的特点,基因谱介于初始T细胞和中央记忆T细胞之间.记忆性干细胞样T细胞具有自我更新和长期存活能力,这些特点使它们成为肿瘤领域关注的热点.本文将简述记忆性干细胞样T细胞的一些特点,以及它们在肿瘤治疗中的应用.     

Cell of origin of lung cancer

Lung cancer is a devastating disease and a major therapeutic burden with poor survival rates. The discovery of rare cells with stem cell‐like properties in solid tumours is emerging as an important area of cancer research and may help explain the resistance of these tumours to current therapeutics. Despite rapid developments in cancer stem cell research in other solid tumours, progress in the lung has been hampered by an incomplete understanding of the epithelial stem cell hierarchy, the heterogeneity of disease and the lack of a suitable in vivo transplantation model to assess stem cell behaviour. In this review we critically discuss what is currently known about the role of normal stem cells and cancer‐initiating cells in lung tumour development, and briefly discuss strategies aimed at advancing the field of lung stem cell biology, with an emphasis on the design and manipulation of state‐of‐art mouse models.     

苯乙酸钠处理的胃癌细胞对LAK细胞杀伤的敏感性

为了探讨分化诱导剂苯乙酸钠是否能够增强ＬＡＫ细胞的治疗效果，作者采用ＭＴＴ法分析了苯乙酸钠处理的胃癌细胞对ＬＡＫ细胞杀伤敏感性的变化。结果表明：苯乙酸钠处理的胃癌细胞ＭＫＮ－２８和ＭＫＮ－４５对ＬＡＫ细胞的杀伤敏感性均增强。另外，苯乙酸钠处理的胃癌细胞粘附ＬＡＫ细胞的活性亦增强。因此，苯乙酸钠处理的胃癌细胞对ＬＡＫ细胞杀伤敏感性的增强可能在于其增强了效靶细胞之间的识别和结合；苯乙酸钠与ＬＡＫ细胞相结合有可能增强ＬＡＫ细胞治疗肿瘤的效果。     

Pancreatic cancer microenvironment

Pancreatic ductal adenocarcinoma remains an extremely aggressive malignancy that is virtually therapy-resistant and has therefore one of the worst prognoses of all human cancers. The focus of research, which had been placed mostly on genetic and epigenetic alterations of the cancer cells themselves, has shifted gradually towards the microenvironment. The cancer microenvironment consists of various components, including fibroblasts, endothelial cells, immune cells, and endocrine cells, that interact with each other and the cancer cells in a complex fashion. This interplay has implications for pancreatic cancer cell growth, migration and invasion, angiogenesis, and immunological recognition of cancer cells. Evidence is accumulating that the cancer microenvironment plays an active role in disease progression, and efforts are being made to target this interplay between cancer cells and host cells to improve the outcome of this deadly disease.     

Tumor-specific T Cells which Form Clusters with Dendritic Cells and Tumor Cells and Deliver Macrophage-activating Factors

T cells prepared from tumor (Meth A)-bearing mice were cocultured with homologous tumor cells and splenic dendritic cells to enrich tumor-specific T cells by the separation of clusters. T blasts generated from clusters were capable of inhibiting the in vivo tumor cell growth. The culture supernatant of clustering cells (CLSN) was effective in activating macrophages (MØ) to be cytostatic and cytocidal against tumor cells. Moreover, it was found that CLSN contains at least 3 distinct factors; one was identified as interferon-γ (IFN-γ), and the others are so far unidentified, but one acts synergistically with IFN-γ, possibly as the second signal, and the other cooperates with lipopolysaccharide but not with IFN-γ. We propose that the tumor-specific T cells secrete soluble mediators which cooperate with each other in MØ activation against tumor cells.     

一种简单γδT细胞扩增培养方法及其抗肿瘤生物学功能研究

目的 :建立一种特异快速地诱导扩增人外周血γδT细胞的方法 ,并比较了γδT细胞 ,NK和LAK细胞的抗肿瘤的生物学特性。方法 :收集用不同单抗分别包被粘附的细胞 ,然后通过MACS细胞分选仪的分选 ,获取的细胞进行细胞增殖动力学、细胞表型、细胞杀伤活性的测定以及单抗阻断效应的分析。结果 :经MACS分离得到的γδT细胞 ,培养 2周后细胞数扩增 6 0 0～ 80 0倍 ,CD3,CD8和γδ细胞表达阳性率分别是 72 .2 9% ,5 8.0 2 %和 6 5 .98%。γδT细胞对NK敏感细胞K5 6 2以及NK不敏感细胞Raji和XG 7这 3种不同靶细胞均有较高的杀伤率 ,分别为 35 .98% ,5 2 .2 7%和 6 9.0 8% ;NK细胞对此 3种细胞的杀伤率分别是 45 2 1% ,12 .34%和 11.94% ;LAK细胞的杀伤率分别为 44 .0 1% ,2 9.2 7%和 2 5 .6 8%。γδT细胞对经MHC Ⅰ类单抗阻断前后的K5 6 2 ,Raji和XG 73种靶细胞的杀伤率无明显改变。结论 :γδT细胞、NK细胞和LAK细胞都具有一定的非特异性杀伤肿瘤细胞的作用 ;γδT细胞对MHC Ⅰ类单抗阻断后的肿瘤细胞的杀伤无明显变化。提示γδT细胞较NK细胞和LAK细胞有更广泛的抗瘤谱     

细胞因子诱导的杀伤细胞在肿瘤治疗中的作用

细胞因子诱导的杀伤（CIK）细胞是用于肿瘤免疫治疗的主要免疫活性细胞。相关基础研究显示其对多种血液及实体肿瘤有显著杀伤活性,大量临床试验证实其对肿瘤患者有明确疗效且不良反应轻微。CIK细胞治疗肿瘤是一种前景广阔且安全的方法,因此建立规范的培养和应用标准十分重要。     

CIK细胞在癌症患者外周血中水平及影响因素的研究

目的 :分析CIK细胞在癌症患者外周血与年龄、性别、疾病种类、分期的关系。方法 :采用流式细胞术 (FCM)对 145例实体瘤患者外周血中的CIK细胞 (CD+ 3 CD+ 56)及NK细胞 (CD-3 CD+ 56)进行标记分析。结果 :在中、晚期实体瘤患者中 ,CIK细胞表达呈高水平 ,CIK细胞占外周血淋巴细胞总数的百分比为 (9 4 5± 5 4 4%) ,在早期表达水平不高 ,为 (3 2 2±2 2 9%) ,两组差异有显著性 (P <0 0 1) ;而年龄、性别、疾病种类并不影响其在外周血中的水平。NK细胞水平不如CIK细胞明显 ,两者无相关性 (P >0 0 5 )。结论 :CIK细胞在中、晚期恶性肿瘤外周血中有较高的表达水平 ,发挥其重要的抗肿瘤细胞免疫作用 ,并不受患者的年龄、性别及疾病种类的影响。     

自体CIK、DC-CIK与半合子DC-CIK抗肿瘤免疫反应的比较研究

Background and Objective:Cytokine-induced killer(CIK) cells and autologous dendritic cells-CIK(DCCIK) cells co-cultured with autologous dendritic cells(DCs) and CIK cells are commonly used for immunotherapy recently.We compared the anti-tumor immune response of CIK cells,autologous DC-CIK cells,and semi-allogeneic DC-CIK cells to explore a more effective anti-tumor adoptive immunotherapy approach.Met hods:Peripheral monocytes were isolated from patients with renal carcinoma,lung cancer,or maxillar...     

Pilot study on the interaction between B16 melanoma cell-line and bone-marrow derived mesenchymal stem cells

Bone-marrow derived mesenchymal stem cells (BMSCs) have the potential to differentiate into osteocytes, chondrocytes, adipocytes and endothelial cells. The interaction between BMSCs and epithelial tumor cell was enhanced on proliferation. Our previous study had shown that BMSCs maybe participate in angiogenesis in melanoma in vivo. The aim of this study was to investigate the interaction between B16 melanoma cells and BMSCs in vitro, the mechanism of BMSCs participating in melanoma angiogenesis in vivo is unclear, so a co-culture system containing BMSCs and B16 melanoma cells, based on transwell indirect model, was established, and the interaction between BMSCs and B16 melanoma cells was studied in vitro. In our study, BMSCs were generated out of bone marrow from C57 mouse, isolated BMSCs were positive for the markers CD105, CD90, CD73, CD44 and CD166 and negative for endothelial markers, which acquired endothelial phenotype (including the expression of VEGFR-1, VEGFR-2, Factor VIII) after co-culture with B16 melanoma cells; at the same time, B16 melanoma cells also up-regulated the expression of VEGF-a, VEGFR-1, VEGFR-2 and Factor VIII. The proliferation rate of B16 melanoma cells and BMSCs were also found to be increased. We could show the differentiation of BMSCs into cells with phenotypic features of endothelial cells. BMSCs promoted proliferation of tumor cells and improved the microenvironment in tumor. Our study suggests that the BMSCs may play an important role in tumor angiogenesis.     

Ovarian tumor initiating cell populations persist following paclitaxel and carboplatin chemotherapy treatment in vivo

Development of recurrent platinum resistant disease following chemotherapy presents a challenge in managing ovarian cancer. Using tumors derived from genetically defined mouse ovarian cancer cells, we investigated the stem cell properties of residual cells post-chemotherapy. Utilizing CD133 and Sca-1 as markers of candidate tumor initiating cells (TIC), we determined that the relative levels of CD133⁺ and Sca-1⁺ cells were unaltered following chemotherapy. CD133⁺ and Sca-1⁺ cells exhibited increased stem cell-related gene expression, were enriched in G₀/G₁-early S phase and exhibited increased tumor initiating capacity, giving rise to heterogeneous tumors. Our findings suggest that residual TICs may contribute to recurrent disease.     

Formation of solid tumors by a single multinucleated cancer cell

BACKGROUND:

Large multinucleated cells (MNCs) commonly exist in tumorigenic cancer cell lines that are used widely in research. However, the contributions of MNCs to tumorigenesis are unknown.

METHODS:

In this study, MNCs were characterized in the murine fibrosarcoma cell line UV‐2237 in vitro and in vivo at the single‐cell level.

RESULTS:

The authors observed that MNCs originated from a rare subpopulation of mononuclear cells and were positive for a senescent marker, β‐galactosidase. In addition, MNCs were responsible for the majority of clonogenic activity when cultured in hard agar; they were more resistant to chemotherapeutic agents than mononuclear cells; they could undergo asymmetric division (producing mononuclear cells) and self‐renewal in vitro and in vivo; and, most important; a single MNC produced orthotopic, subcutaneous tumors (composed mainly of mononuclear cells) that gave rise to spontaneous lung metastases in nude mice.

CONCLUSIONS:

The current results indicated that the growth of MNCs may be arrested under stress and that MNCs are highly resistant to chemotherapy and can generate clonal, orthotopic, metastatic tumors. Cancer 2011. © 2011 American Cancer Society.     

胎儿LAK细胞治疗中晚期肝癌的疗效观察

自1989年1月至1991年10月,我们应用6~7个月龄的胎儿脾脏细胞和白细胞介素-2(IL-2)共培养3天后制备成胎儿LAK细胞,静脉输注或肝动脉插管灌注治疗中晚期肝癌18例.结果疼痛减轻16例,占88.9%;食欲增加14例,占77%;临床完全缓解2例,占11%;部分缓解14例,占77%;总有效率占88.9%(16/18).而且LAK细胞治疗后患者周围血OKT_4比率升高,OKT_8比率下降,说明治疗后患者免疫功能有一定改善.结果提示:①胎儿脾脏含有丰富的LAK前身细胞;②胎儿LAK细胞治疗中晚期肝癌有明显疗效.     

Immune activation by combination human lymphokine-activated killer and dendritic cell therapy

Background:

Optimal cellular immunotherapy for cancer should ideally harness both the innate and adaptive arms of the immune response. Lymphokine-activated killer cells (LAKs) can trigger early innate killing of tumour targets, whereas long-term adaptive-specific tumour control requires priming of CD8+ cytotoxic lymphocytes (CTLs) following acquisition of tumour-associated antigens (TAAs) by antigen-presenting cells such as dendritic cells (DCs). As DCs stimulate both innate and adaptive effectors, combination cell therapy using LAKs and DCs has the potential to maximise anti-tumour immune priming.

Methods:

Reciprocal activation between human clinical grade LAKs and DCs on co-culture, and its immune consequences, was monitored by cell phenotype, cytokine release and priming of both innate and adaptive cytotoxicity against melanoma targets.

Results:

Co-culture of DCs and LAKs led to phenotypic activation of natural killer (NK) cells within the LAK population, which was associated with increased production of inflammatory cytokines and enhanced innate cytotoxicity against tumour cell targets. The LAKs reciprocally matured DCs, and the combination of LAKs and DCs, on addition of melanoma cells, supported priming of specific anti-tumour CTLs better than DCs alone.

Conclusion:

Clinical-grade LAKs/DCs represents a practical, effective combination cell immunotherapy for stimulation of both innate and adaptive anti-tumour immunity in cancer patients.     

DC-CIK联合化疗对乳腺癌患者Treg细胞表达及预后的影响

目的：探讨树突状细胞（DC）与细胞因子诱导的杀伤细胞（CIK）联合化疗对乳腺癌患者Treg 细胞表达和预后的影响。方法试验组42例患者术后接受 DC-CIK 细胞联合化疗,对照组38例患者仅接受单纯化疗,比较两组患者治疗后外周血中 Treg 细胞表达量,比较两组近期疗效、无进展生存期（PFS）、总生存期（OS）以及治疗后的生命质量。结果治疗后1周试验组患者外周血中 Treg 细胞表达量明显低于对照组[（11．37±1．10）％∶（14．25±0．95）％],治疗后2周试验组患者外周血中 Treg 细胞表达量明显低于对照组[（7．94±1．12）％∶（9．14±1．21）％],差异均有统计学意义（t ＝12．470,P ＝0．000;t ＝4．606,P ＝0．000）。试验组临床有效率与对照组比较（47．62％∶44．74％）,差异无统计学意义（χ2＝0．07,P ＝0．80）;试验组疾病控制率明显高于对照组（80．95％∶60．53％）,差异有统计学意义（χ2＝4．06,P ＝0．04）。试验组患者的 PFS[（7．50±1．45）个月∶（5．50±1．52）个月]和 OS[（13．50±3．20）个月∶（11．50±3．25）个月]均明显长于对照组,差异均有统计学意义（t ＝6．021,P ＝0．000;t ＝2．771,P ＝0．007）。治疗后试验组患者在躯体功能（72．85±12．01∶57．42±13．07）、情绪功能（68．45±9．97∶44．79±9．15）、认知功能（67．54±9．95∶62．37±10．34）、社会功能（65．72±12．17∶49．37±8．45）和总体生命质量（71．43±11．50∶59．48±12．45）方面的评分均明显高于对照组,差异均有统计学意义（t ＝5．503,P ＝0．000;t ＝11．020,P ＝0．000;t ＝2．278,P ＝0．025;t ＝7．032,P ＝0．000;t ＝4．463,P ＝0．000）。结论DC-CIK 联合化疗能够显著降低乳腺癌患者外周血中 Treg 细胞的表达,改善免疫抑制情况,同时提高疗效,延长 PFS,提高患者的生命质量。     

肿瘤干细胞的研究进展

肿瘤干细胞是存在于肿瘤中的一小部分具有干细胞性质的细胞群,具有高度的致瘤性和耐药性。同正常干细胞一样,肿瘤干细胞具有无限的自我更新和多向分化的潜能,使肿瘤在体内不断扩大或形成新的肿瘤,导致肿瘤复发和转移。肿瘤干细胞的研究有助于认识和理解肿瘤发生发展的机制,指导肿瘤的临床治疗。     

A-NK细胞抗肝癌Walker256瘤株的实验研究

目的:观察A-NK细胞的体外生长与增殖,以及杀伤肿瘤细胞的能力,研究A-NK细胞局部注射的体内抗肿瘤作用。方法:用淋巴细胞分离液分离单个核细胞(PBMC),培养LAK细胞和A-NK细胞,分别将LAK细胞、A-NK细胞和Walker-256瘤株细胞接种于培养板中,培养24h后,用四甲基偶氮唑盐(MTT)方法测定吸光度,计算肿瘤杀伤率。复制鼠肝癌模型,分别将LAK、A-NK细胞经肝动脉注入鼠肝癌模型中,观察两组动物的生存时间。结果:A-NK细胞的增殖明显快于LAK细胞(P<0.05),A-NK细胞对肿瘤细胞的杀伤活性比LAK细胞强(P<0.01),A-NK细胞能显著延长肝癌动物模型的生存期,与LAK细胞比较,具有显著性差异(P<0.01)。结论:A-NK细胞具有增殖快,抗瘤活性强,在体内能抑制肿瘤生长,延长荷瘤动物的生存期。     

人类羊膜上皮细胞与间充质干细胞的免疫调节功能

 成体干细胞是一种具有自我更新和多向分化能力的细胞,在组织工程、基因治疗和细胞移植领域具有较好的应用前景。近年研究显示,人类羊膜中也存在两种干细胞：羊膜上皮细胞（HAEC）和羊膜间充质干细胞（hAMSC）,由于具有来源广泛、取材方便、多向分化潜能以及免疫原性低等优点,在细胞移植中表现出免疫调节等作用,且可作为细胞组织工程种子,正逐渐成为干细胞研究领域的热点之一。现就两种羊膜细胞生物学特性、免疫调节机制以及临床运用的潜在价值作一综述。