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[目的]研究树突状细胞(DC)联合细胞因子诱导或未诱导的杀伤细胞(CIK)或淋巴因子激活的杀伤细胞(LAK)对结肠癌细胞株SW480的杀伤活性。提供DC联合CIK或LAK治疗结肠癌的实验依据。[方法]取人外周血分离出单个核细胞(PBMNC),诱导生成DC、CIK、LAK细胞;流式细胞仪检测DC经SW480肿瘤抗原冲击后的表型变化;以CIK+DC细胞、CIK细胞、LAK+DC细胞及LAK细胞作为效应细胞,SW480为靶细胞,以15∶1、30∶1、45∶1为效靶比,LDH释放法测定细胞杀伤试验活性;ELISA检测杀伤试验中干扰素γ(IFN-γ)、白细胞介素2(IL-2)、IL-12、IL-17的分泌水平。[结果]流式细胞仪检测DC经SW480肿瘤抗原冲击后,其表面分子HLA-DR、CD40、CD80和CD86表达分别平均为90.23%、73.68%、85.96%、57.55%,与未经肿瘤抗原冲击DC比较,DC成熟的表面标志分子表达明显增加(P<0.01)。相同效靶比下,CIK+DC细胞组对SW480的杀伤作用最强,明显高于其他细胞组(P<0.01);CIK+DC细胞组在效靶比为45∶1时,杀伤活性最强(P<0.01);单独CIK细胞组的杀伤活性明显高于LAK+DC细胞组(P<0.01);LAK+DC细胞组的杀伤活性明显高于单独LAK细胞组(P<0.01)。效靶比为45:1时,各杀伤试验细胞组上清液中IFN-γ、IL-2、IL-12、IL-17的分泌量,CIK+DC细胞组的IFN-γ、IL-12的分泌量显著高于其他细胞组(P<0.05);LAK+DC、单独LAK细胞组IL-2的分泌量明显高于CIK+DC、单独CIK细胞组(P<0.05);单独CIK细胞组IFN-γ的分泌量明显高于LAK+DC、单独LAK细胞组(P<0.05)。[结论]CIK+DC细胞组对SW480的杀伤活性明显强于单独CIK、LAK+DC组、单独LAK细胞组。其机制可能是,SW480抗原致敏的DC分泌IFN-γ、IL-12等刺激、诱导CIK细胞的活化和增殖,明显增强CIK细胞杀伤SW480的活性。
[Objective] To study the killing activity of dendritic cells (DC) combined with cytokine-induced or uninduced killer (CIK) or lymphokine-activated killer (LAK) on colon cancer cell line SW480. Provide experimental evidence of DC combined with CIK or LAK in the treatment of colon cancer. [Methods] Peripheral blood mononuclear cells (PBMNC) were isolated and induced to produce DC, CIK and LAK cells. Flow cytometry was used to detect the phenotypic changes of DCs exposed to SW480 tumor antigen. CIK + DC cells and CIK cells , LAK + DC cells and LAK cells as effector cells, and SW480 as target cells. The cytotoxic activity was determined by LDH release assay at 15:1, 30:1 and 45:1 respectively. γ (IFN-γ), interleukin 2 (IL-2), IL-12, IL-17 secretion levels. [Results] The results of flow cytometry showed that the expression of HLA-DR, CD40, CD80 and CD86 on the surface of SW480 cells was 90.23%, 73.68%, 85.96% and 57.55% on the average, respectively. DC, the expression of mature DCs was significantly increased (P <0.01). At the same target ratio, CIK + DC cells had the strongest cytotoxic effect on SW480 cells (P <0.01), while CIK + DC cells had the strongest cytotoxic activity at 45:1 (P <0.01). The killing activity of CIK cells alone was significantly higher than that of LAK + DC cells (P <0.01). The killing activity of LAK + DC cells was significantly higher than that of LAK cells alone (P <0.01). The ratio of IFN-γ, IL-2, IL-12 and IL-17 in supernatant of cytotoxic groups was significantly higher than that in CIK + DC cells (P <0.05). The secretion of IL-2 in LAK + DC and LAK alone group was significantly higher than that in CIK + DC and CIK cells alone group (P <0.05) The secretion of IFN-γ was significantly higher than that of LAK + DC and LAK cells alone (P <0.05). [Conclusion] The killing activity of SW480 in CIK + DC cell group was obviously stronger than that in CIK, LAK + DC group and LAK cell alone groups. The mechanism may be, SW480 antigen-primed DC secretion of IFN-γ, IL-12 and other stimuli, induce CIK cell activation and proliferation, significantly enhanced CIK cell killing SW480 activity.