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食管癌细胞与源自脐带血的树突状细胞融合体的抗肿瘤活性

Antitumor activity of a fusion of esophageal carcinoma cells with dendritic cells derived from cord blood.

作者信息

Guo Guanghua, Chen Suzuan, Zhang Juan, Luo Lili, Yu Jing, Dong Hongmei, Xu Hong, Su Zhongjing, Wu Libiao

机构信息

Department of Gastroenterology, The First Affiliated Hospital of Shantou University Medical College, Shantou 515041, Guangdong Province, China.

出版信息

Vaccine. 2005 Nov 1;23(45):5225-30. doi: 10.1016/j.vaccine.2005.07.080. Epub 2005 Aug 18.

Abstract

The aim of this experiment was to develop a cytotoxic cancer vaccine (EC109-DC) prepared by fusions of esophageal carcinoma cells with dendritic cells derived from cord blood and to study the biological characteristics and resultant induction of antitumor immunity. CD34+ hematopoietic stem cells were isolated from cord blood using a CD34+ Progenitor Cell Isolation Kit by magnetic cell sorting system (MACS). CD34+ cells were incubated with rhGM-CSF, rhTNF-alpha and rhSCF for 2 weeks as DC (dendritic cells), and then by PEG-3600 to fuse with an esophageal carcinoma cell line. Selection with MACS marked with HLA-DR MicroBeads generated EC109-DC. Phenotypes and proliferation were analyzed by flow cytometry and cell culture in vitro. The lymphocyte proliferation reaction and CTL cytotoxicity were examined by MTT assay. The EC109-DC cells could proliferate slowly in vitro and highly expressed CD80, CD83 and CD86. The lymphocyte proliferation reaction and specific cytotoxicity against EC109 induced by EC109-DC cells were significantly higher than in control groups (p < 0.05). EC109-DC cells obtained by PEG fusion acquired the immuno-stimulating phenotype and could significantly stimulate the lymphocyte proliferation reaction and CTL activity. The results of this research provide the basis for materials to develop the DC-based vaccine against esophageal carcinoma.

摘要

本实验旨在研制一种通过食管癌细胞与脐血来源的树突状细胞融合制备的细胞毒性癌疫苗(EC109-DC),并研究其生物学特性及由此诱导的抗肿瘤免疫。使用CD34+祖细胞分离试剂盒通过磁性细胞分选系统(MACS)从脐血中分离CD34+造血干细胞。将CD34+细胞与重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)、重组人肿瘤坏死因子-α(rhTNF-α)和重组人干细胞因子(rhSCF)一起培养2周以获得树突状细胞(DC),然后通过聚乙二醇3600(PEG-3600)与食管癌细胞系融合。用标记有HLA-DR微珠的MACS进行筛选得到EC109-DC。通过流式细胞术和体外细胞培养分析其表型和增殖情况。通过MTT法检测淋巴细胞增殖反应和细胞毒性T淋巴细胞(CTL)的细胞毒性。EC109-DC细胞在体外能缓慢增殖,高表达CD80、CD83和CD86。EC109-DC细胞诱导的淋巴细胞增殖反应和对EC109的特异性细胞毒性显著高于对照组(p < 0.05)。通过PEG融合获得的EC109-DC细胞具有免疫刺激表型,能显著刺激淋巴细胞增殖反应和CTL活性。本研究结果为开发基于DC的食管癌疫苗提供了物质基础。

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