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背景:冷冻免疫是近年来倍受关注的课题,但冷冻对细胞免疫功能的影响一直缺乏深入的研究,特别是在细胞凋亡和树突状细胞方面。目的:观察体外培养的肺癌NCI-H446细胞经氩氦刀冻融处理后细胞形态和细胞表面免疫表型的变化,及其能否有效激发骨髓树突状细胞产生特异性抗瘤效应。设计、时间及地点:细胞免疫水平的对照实验,于2002-08/2003-04在解放军海军总医院血液实验室及解放军军事医学科学院细胞与基因治疗中心完成。材料:小细胞肺癌细胞系NCI-H446细胞购自上海科学院细胞库。方法:取体外培养的肺癌NCI-H446细胞经氩氦刀处理后,在体外树突状细胞培养过程中,加入冻融的肺癌细胞,分别观察肺癌细胞的形态结构、混合淋巴细胞反应及细胞毒T淋巴细胞杀癌细胞效应。主要观察指标:培养树突状细胞的细胞形态和细胞表面免疫表型变化,培养细胞的混合淋巴细胞反应结果,培养细胞的凋亡情况。结果:骨髓树突状细胞体外培养后,符合树突状细胞特征。在培养过程中加入冻融的肺癌细胞,不影响树突状细胞细胞表面抗原的表达,混合淋巴细胞反应增强,细胞毒T淋巴细胞可引起肺癌细胞凋亡。结论:经氩氦刀处理的肺癌细胞溶解,胞膜不完整,体外能有效激发骨髓树突状细胞产生特异性抗瘤效应。
Background: Freezing immunity is a topic of great concern in recent years. However, the effects of freezing on cellular immune function have not been studied deeply, especially in the aspects of apoptosis and dendritic cells. OBJECTIVE: To observe the change of cell morphology and cell surface immunophenotype in NCI-H446 cells cultured in vitro by argon-helium cryoablation and whether it can effectively stimulate bone marrow dendritic cells to produce specific anti-tumor effects. DESIGN, TIME AND SETTING: The control experiment of cellular immune level was performed at the Blood Laboratory of People’s Liberation Army Navy General Hospital and the Center for Cell and Gene Therapy, Academy of Military Medical Sciences, People’s Liberation Army from August 2002 to April 2003. Materials: Small cell lung cancer cell line NCI-H446 cells were purchased from Shanghai Academy of Sciences cell bank. Methods: The lung cancer NCI-H446 cells were treated with argon-helium knife in vitro. During the process of dendritic cell culture in vitro, freeze-thaw lung cancer cells were added to observe the morphology, mixed lymphocyte reaction and cytotoxicity T lymphocytes kill cancer cells. MAIN OUTCOME MEASURES: Cell morphology and cell surface immunophenotypic changes of cultured dendritic cells, mixed lymphocyte reaction results of cultured cells, and apoptosis of cultured cells. RESULTS: Bone marrow dendritic cells cultured in vitro were in accordance with the characteristics of dendritic cells. The addition of freeze-thawed lung cancer cells during culture does not affect the expression of dendritic cell surface antigens, and the mixed lymphocyte reaction is enhanced. Cytotoxic T lymphocytes can cause lung cancer cell apoptosis. CONCLUSION: The lung cancer cells treated with argon-helium knife are lysed and the cell membrane is incomplete, which can effectively stimulate bone marrow dendritic cells to produce specific anti-tumor effect in vitro.