【摘要】 目的 体外观察胰腺癌(PC)患者树突状细胞(DC)增生成熟,DC分泌的细胞因子及DC诱导免疫效应细胞分泌的细胞因子对胰腺癌细胞PC3的凋亡作用。方法 用淋巴细胞分离液分离获取外周血单核细胞(PBMC),贴壁法获取DC和去DC的单核细胞(即免疫效应细胞),分别用粒/巨噬细胞集落刺激因子(GM-CSF)1 000 U/ml,人白细胞介素-4(IL-4)500 U/ml,肿瘤坏死因子(TNF)-α 500 U/ml、PC3肿瘤相关(TAA)和人 IL-2 100 U/ml培养 DC和免疫效应细胞,观察 DC生长状况,检测DC表型(CD1a、CD80、CD83、CD86)及DC培养上清液对PC3细胞的凋亡作用。结果 体外多种细胞因子和肿瘤相关抗原能有效引起胰腺癌患者 DC增殖 0.5×10~5～1.0×10~5个/ml,高表达CD80、CD83、CD86。DC培养上清液和DC混合免疫效应细胞培养上清液均能有效地引起PC3凋亡。结论 DC在胰腺癌免疫治疗中有重要作用。更多还原

【Abstract】 Objective To observe dendritic cell (DC) proliferation and maturation, and the effectsof cytokines secreted by DC and the cytokines by DC-induced immunoeffect cells on PC3 apoptosis ex vi-vo. Methods The DC and mononuclear cells without DC (namely immunoeffect cells) were isolated andpurified from peripheral blood mononuclear cells (PBMC) in the patients with pancreatic cancer. The DCwere cultured in the presence of granulocyte/ macrophage colony stimulating factor (1 000 U/ml), inter-leukin-4 (500 U/ml), tumor necrosis factor-α(500 U/ml) and extracted tumor-associated antigen(TAA) from human pancreatic cancer cell line PC3, and the immunoeffect cells were culture with inter-leukin-2 (100 U/ml), respectively. 5 or 6 days later, DC and immunoeffect cells were cocultured for twodays ex vivo. The effects of DC phenotypes (CD1a, CD80, CD83 and CD86) and the culture supernatantof DC on the apoptosis of PC3 were determined. Results In vitro cytokines and tumor related antigenscould effectively result in the DC amplification to 0. 5×10~5 ～1. 0×10~5 DC/ml and high expression ofCD80, CD83 and CD86. Both the culture supernanant of DC and that of DC in combination with immu-noeffect cells could induce the apoptosis of PC3. conclusion DC might play an important role in the im-munotherapy of pancreatic cancer.更多还原