树突状细胞与细胞因子诱导的杀伤细胞共培养抗非霍奇金淋巴瘤作用研究  

Effect of dendritic cells co-cultured with cytokine induced killer cells on cytotoxicity against non Hodgkin＇s lymphoma cell

作　　者：刘小兰[1] 关涛[2] 苏丽萍[2] 

机构地区：[1]山西医科大学研究生院,太原030001 [2]山西医科大学附属肿瘤医院血液科

出　　处：《白血病．淋巴瘤》2013年第8期466-469,共4页Journal of Leukemia & Lymphoma

摘　　要：目的研究细胞因子诱导的杀伤细胞（CIK细胞）与树突状细胞（DC）共培养后细胞的增殖活性、细胞因子分泌水平、免疫表型及其对非霍奇金淋巴瘤Raji细胞的体外杀伤作用。方法采集健康人群外周血单个核细胞，于37℃、5％CO2、饱和湿度培养箱中培养2h；收集贴壁细胞用于诱导培养DC，非贴壁细胞诱导培养CIK细胞，分别培养9d；将成熟DC与CIK细胞共同培养6d，以单纯CIK细胞作为对照组。采用四甲基偶氮唑蓝（MTF）比色法检测CIK细胞、DC和CIK共培养细胞对非霍奇金淋巴瘤Raji细胞杀伤活性。同时分别检测第0、3、6、9、12、15天培养细胞的细胞形态、细胞增殖量、细胞因子分泌量及免疫表型受体等指标。结果在细胞培养第12天，DC．CIK细胞与单纯CIK细胞相比，增殖明显[（42．44±2．68）倍比（30．01±2．05）倍]（t=11．64，P〈0．05），白细胞介素（IL）-2、干扰素（IFN）-γ、IL．12及肿瘤坏死因子（TNF）-α分泌量增加[（124．34±12．57）ng／L比（56．32±6．58）ng／L、（496．60±95．32）ng／L比（247．80±69．45）ng／L、（84．92±6．07）ng／L比（24．18±3．31）ng／L、（380．60±45．95）ng／L比（196．61±24．19）ng／L1（t值分别为15．16、6．67、27．78、11．20，均P〈0．05），CD3＋CD；和CD；CD56细胞比例也提高[（71．79±1．73）％比（60．37±3．24）％、（48．54±3．30）％比（33．07±2．22）％]（t=9．83，P〈0．05；t=12．30，P〈0．05），并在同一效靶比下，对Raii细胞具有更强的杀伤活性。结论DC与CIK细胞共培养后可促使CIK细胞成熟、增殖，并提高抗肿瘤作用。Objective To study proliferation, secreted cytokines, immune phenotypes and cytotoxicity on Raji cells by cytokine induced killer （CIK） cells co-cultured with dendritic cells （DC）. Methods The mononuclear cells from peripheral blood of healthy individuals were extracted, then cultured the cells under 5 % CO2 at 37℃ for 2 hours. DC were induced from suspended cells, and CIK cells were from adherent cells. After 9 days of nurturing, two types of cells were mixed.CIK cells were cultured alone as the control. The cytotoxic activity of CIK and DC-CIK cells were detected by MTI＇ assay. The morphologies, proliferation, secreted cytokines, and immune phenotypes of the two cells in day 0, 3, 6, 9, 12, 15 in culture were monitored. Results In day 12 in culture, comparing with CIK cells, DC-CIK cells significantly enhanced the proliferation rate [（42.44-±2.68） fold vs （30.01-±2.05） fold] （t = 11.64, P 〈 0.05） and had increased IL-2, IFN-% IL-12 and TNF-~ secretion [（124.34-±12.57） ng/L vs （56.32-±6.58） ng/L, （496.60±95.32） ng/L vs （247.80-± 69.45） ng/L, （84.92±6.07） ng/L vs （24.18±3.31） ng/L, （380.6±45.95） ng/L vs （196.61±24.19） ng/L] （t = 15.16, P〈 0.05; t = 6.67, P〈 0.05; t = 27.78, P〈 0.05; t = 11.20, P〈 0.05）, and there were more CD3 CD; cells and CD3 CD56 cells in the co-culture [（71.79±1.73） % vs （60.37±3.24） %, （48.5±3.30） % vs （33.07-±2.22） %] （t = 9.83, P 〈 0.05; t = 12.30, P 〈 0.05）, and DC-CIK cells had a significantly increased cytotoxicity on Raji cells in vitro at the same ratio of effector cells to target cells. Conclusion CIK cells have higher proliferation rate and cytotoxicity against Raji cells when co-cultured with DC.

关 键 词：树突状细胞 细胞因子诱导的杀伤细胞 淋巴瘤 非霍奇金 过继细胞免疫治疗 

分 类 号：R733.1[医药卫生—肿瘤]