摘要
[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。
Abstract： [Objective]To compare the cytotoxic effect of dendritic cells （DC） co-cultured with cytokine- induced killer cells （CIK） or lymphokine-activated killer cells （LAK） on SW480 cell line, so as to provide the experimental data for CIK cells co-cultured with DC or LAK cells co-cultured with DC in the treatment of cancer. [Methods]Human peripheral blood mononuclear cells （PBMNCs） separated from healthy donors were induced to CIK, LAK and DC. After treated with colon cancer cell lysate antigen, the phynotypes of DC were detected by FCM. CIK＋DC, CIK cells, LAK＋DC and LAK cells acted as the effector cells. SW480 cells acted as the target cells. The eytotoxic activity was detected by LDH release assay at 15.1,30. 1,45 ： 1 ratios of effector cells to target cells. The secretory amount of IFN-γ, IL-2, IL-12, and IL-17 was detected by ELISA in the collected supernatants of every reaction groups. [Results]The phenotyp expression of HLA-DR,CD40, CDS0 and CD86 in DC pulsed with colon cancer lysate antigen increased to 90. 23%, 73.68%,85.96% and 57. 55% respectively. DC became mature after pulsed with colon cancer lysate anti- gen. At the same ratio of effector cells to target cells,CIK＋DC group showed the strongest cytotoxic effect on SW480 than the other groups （P〈0.01）,especially at the ratio of 45： 1. The cytotoxic effect on SW480 of CIK group was stronger than LAK＋DC group （P〈0.01）,and the cytotoxic effect to SW480 of LAK＋ DC group are stronger than LAK group （P〈0. 01）. At the ratio of 45：1 of effector ceils to target cells, the secretory amount of IFN-γ, IL-2, IL-12, IL-17 was evaluated. The secretory amount of IFN-7 and IL-12 was higher in CIK＋DC group than the other groups （P〈0.05）. The secretory amount of IL-2 in LAK＋DC group and LAK group was higher than the CIK＋DC group and CIK group （P〈0. 05）. The secretory a- mount of IFN-γ in CIK group are higher than LAK＋DC group and LAK group（P〈0. 05）. [Conclusion] CIK＋DC group had a ob
出处
《中国中西医结合消化杂志》
CAS
2013年第3期122-126,共5页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
基金
台州市科学技术局科学技术研究项目(20111ky1102)
