淋巴细胞体外杀伤小鼠神经母细胞瘤实验观察  

Lethality of Spleen Lymphocyte in Vitro to Neuroblastoma Cell N2a of Mice

作　　者：王政[1] 罗德炎[2] 段跃强[2] 王希良[2] 

机构地区：[1]中国人民解放军总医院小儿外科,北京100853 [2]军事医学科学院微生物流行病研究所免疫学研究室,北京100071

出　　处：《医学临床研究》2010年第3期385-388,共4页Journal of Clinical Research

基　　金：国家自然基金资助项目（30170853）

摘　　要：【目的】建立体外小鼠神经母细胞瘤细胞和淋巴细胞共培养的细胞模型，通过检测不同免疫学指标来衡量淋巴细胞杀伤肿瘤细胞能力。【方法】从正常C57BL／6小鼠体内分离脾淋巴细胞，调节到适当的浓度，将神经母细胞瘤的细胞株（N2a）和新鲜分离的脾淋巴细胞共培养。在倒置显微镜下观察脾淋巴细胞形态，同时分离培养上清，用ELISA方法检测IL-6、IFN-r、TNF-α含量。MTT方法检测共培养体系中存活细胞状况，用来反映脾淋巴细胞体外杀伤神经母细胞瘤细胞的能力。【结果】体外共培养模型显示，脾淋巴细胞具有强大的杀伤N2a肿瘤细胞的能力，37℃5％C02培养48h后，在4—1×10^6／mL脾淋巴细胞浓度下无肿瘤细胞生长。细胞培养上清中可以检测到高浓度的IL-6（1849pg／mL）和中浓度的TNF-α（137pg／mL），却检测不到IFN-r。【结论】脾淋巴细胞具有很强体外杀伤N2a肿瘤细胞能力，IL-6可能发挥着比TNF-α更重要作用，这为临床上神经母细胞瘤手术后的免疫治疗提供了新的思路。[Objective]To establish coculture model of spleen lymphocyte in vitro and N2a neuroblastoma cell in order to measure and deduce spleen lymphocyte＇s lethality to neuroblastoma cell. [Methods]The spleen was isolated from the wild-type C57BL/6 mice without any dealings to prepare spleen lymphocyte with conven- tional method. Appropriate cell concentration was adjusted. And then neuroblastoma cell N2a was cocultured with spleen lymphocytes. The morphological characteristics under optical microscope were observed. The anti- tumor antibody titer and cytokines contents（IL-6, IFN-r, TNF-α） were detected by ELISA. The MTT assay was used to measure the tumor-killed ability of spleen lymphocyte. [Results]In vitro coculture model, spleen lymphocyte displayed the strong tumor-killed ability. No neuroblastoma cell N2a was alive which was meas- ured by microscope and MTT assay when it was cocultured with 4-1× 10^6/mL spleen lymphocyte 48h in 37℃ 5% CO2 condition. In the coculured cell supernatant, high concentration of IL-6 （1849 pg/mL） and TNF-a （137 pg/mL） was detected, but IFN-r was not detected. [Conclusion] Spleen lymphocytes isolated from normal mice have strong N2a neuroblastoma-killed ability in vitro. It will be the good guidance for immunotherapy for neuroblastoma after surgery.

关 键 词：神经母细胞瘤 淋巴细胞 小鼠 

分 类 号：R730.264[医药卫生—肿瘤]