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恶性胸腔积液来源树突状细胞对自体肿瘤 浸润性淋巴细胞的作用

Effect of Dentritic Cells from Malignant Pleural Effusions on Tumor Infiltration Lymphocytes

  • 摘要: 目的探讨恶性胸腔积液来源DCs(dentritic cells,DCs)对自体肿瘤浸润性淋巴细胞(tumor infiltration lymphocytes, TILs)增殖及杀伤肿瘤细胞能力的影响。方法用体外培养方法从16例肺癌患者恶性胸腔积液来源分离单个核细胞,再用密度梯度离心辅以免疫磁珠分选细胞,用白细胞介素4(IL-4 )、肿瘤坏死因子-α(TNF-α)、粒-单核细胞刺激因子(GM-CSF)等诱导出DCs,用流式细胞仪和电子显微镜鉴定分离DCs;用IL-2、植物血凝素诱导同一患者胸腔积液单个核细胞中的TILs,用HEA-125磁珠分选纯化肿瘤细胞,3H-TdR渗入法检测DCs对TILs增殖能力;MTT法检测TILs对肿瘤细胞杀伤活性。结果从肺癌患者恶性胸腔积液单个核细胞中可诱导成熟DCs,电镜和光镜观察结果显示,这类DCs具有典型的DC形态,高表达HLA-ABC、HLA-DR、CD86,较高表达CD80、CD54,也表达CD83、CD1a。DCs可明显促进TILs增殖能力(增加约1.7倍)。以TILs本身为对照,DCs激活的TILs对肿瘤细胞杀伤活性明显增加[从(31.80±14.05)%提高到(51.89±13.27)%,P<0.05]。结论肺癌患者恶性胸腔积液单个核细胞中可诱导成熟DCs ,这种DCs可刺激自体TILs扩增,增加TILs杀伤肿瘤细胞的活性。

     

    Abstract: Objective Dentritic cells (DCs) from malignant pleural effusions of patients with lung cancer were isolated and induced.Effect of DCs on proliferation and cytotoxicity of tumor infiltration lymphocytes (TILs) from origin malignant pleural effusions was studied. Methods Pleural effusion mononuclear cells were obtained from 16 patients with lung cancer.Cells were separated by density gradient and magnetic cell sorting system.DCs were induced by interleukin-4 (IL-4), tumor necrosis factor-α(TNF-α), and granulocyte-macrophage colony stimulating.DCs were observed by optical microscope, electronic microscope and flow cytometry respectively.TILs from origin malignant pleural effusions were induced by IL-2 and phytohemagglutinin.The cancer cells were separated with HEA-125 magnetic cell sorting system.The ability of stimulating the proliferation of TILs was detected with 3H-thymidine.The anti-tumor effect of TILs was measured with MTT method. Results The mature DCs from malignant pleural effusions of patients with lung cancer could be induced.The typical morphology in DCs was observed with optical and electronic microscopes.DCs expressed high-level surface phenotypes, including HLA-ABC, HLA-DR, CD86, CD54 and CD83, CD1a.DCs can increase about 1.7 times proliferation of TILs.The cytotoxicity of TILs stimulated with DCs was increased from (31.80±14.05)% to (51.89±13.27)%. Conclusion The mature DCs can be induced from malignant pleural effusions from patients with lung cancer.The DCs can stimulate TILs from the origin malignant pleural effusion and increase its proliferation ability and cytotoxicity to cancer cells.

     

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