活化抗原提呈细胞对人骨肉瘤PL-11细胞凋亡后体外免疫原性的影响  

Influence of activated antigen presenting cells on the immunogenicity of apoptotic human osteosarcoma cell line PL-11

作　　者：吕昌伟[1] 王臻[1] 李立文[1] 彭磊[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,陕西西安710033

出　　处：《第四军医大学学报》2001年第11期971-974,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金!资助 (3 9770 749)

摘　　要：目的　研究人骨肉瘤细胞 PL- 11被诱发凋亡后 ,体外活化的抗原提呈细胞 (APC)对其免疫原性的影响 .方法　1H2 O2 诱发 PL- 11凋亡的浓度和时间筛选 ,流式细胞仪分析 .2正常献血者粗分白细胞悬液离心沉降法得人外周血单个核细胞 ,粘附法分离单核细胞并预激活 ,同时分离得混合淋巴细胞并预培养 .激活单核细胞分别与凋亡细胞多次冻溶物(PL- 11a)和未发生凋亡之 PL- 11的多次冻溶物 (PL- 11u)和淋巴细胞孵育 7d(pl- 11a组及 pl- 11u组 ) ,另设单纯淋巴细胞与凋亡 PL- 11多次冻溶物培养组 (对照组 )和激活单核细胞与淋巴细胞共培养 (MON组 )组 ,计数法测量增殖 .3各组培养的淋巴细胞进行体外杀伤实验 (效靶比 10 0∶ 1) ,改良 MTT(thiazolyl blue)法测量杀伤及抑制率 .结果　 1低于 10 0nmol·L- 1 H2 O2 在 7h之内均无明显诱导凋亡作用 ,于 14h后均诱导凋亡 ,且存在量效差别 ,10 0 nmol· L- 1处理 36 h最明显 .2 PL - 11a组淋巴细胞体外增殖显著 ,PL - 11u组仅有较弱的增殖作用 ,对照组和 MON组均无明显增殖作用 .3pl- 11u组仅有较弱的淋巴细胞杀伤作用 ,pl- 11a组则有明显的增强 ,而对照组及 MON组均无作用 .结论　人骨肉瘤细胞在 H2 O2 诱发凋亡后 ,如经活化 APC吞噬 ,可显著提高其体外免疫原性而引出肿?AIM To study the influence of actived antigen presenting cells (APC) on the in vitro immunogenicity of apoptotic human osteosarcoma PL 11. METHODS ① To select H 2O 2 concentration and working time to induced apoptosis, flow cyte meter (FCM) was implied to analyse ② Periphery blood mononuclear cells (PBMC) were seperated from rurally seperated white cells by density gradient centrifugation, and monocytes was seperated by adhesion and activated. The repeated freeze thaw of apoptotic cells (PL 11a) and unapoptotic cells (PL 11u) were cultured with lymphocytes and activated monocytes for 7 days (pl 11a group and pl 11u group) respectively. Lymphocytes with PL 11a (control group) and lymphocytes with activated monoctyes (MON group) were parallelly cultured, and the proliferation of each group were counted. ③ Lymphocytes from each group were harvested to measure the killing ability in vitro by modified MTT (thiazolyl blue) method. RESULTS ① H 2O 2 wasn't able to induce apoptosis at 100 nmol·L -1 concentration or lessor in 7 h, but could induce apoptosis after 14 h, especially the 100 nmol·L -1 after 36 h. ② Lymphocytes of the PL 11a group increased significently, while that of the PL 11u group increased merely and the control group and MON group have no increase. ③ The pl 11u group showed a low cytotoxicy to osteosarcoma cells, and the pl 11a group had a significent increase ( P <0.01). The control group and the MON group had no cytotoxity ( P >0.05). CONCLUSION Induced apoptosis by H 2O 2, human osteosarcoma cells could increase immunogenicity in vitro through the activated PBMC monocytes, this indicates that the activated monocytes phagocytosed repeated freeze thaw of apoptotic human osteosarcoma will be a new approach for individual tumor vaccine.

关 键 词：H2O2单核细胞 骨肉瘤 细胞凋亡 体外免疫原性 活化抗原提呈细胞 

分 类 号：R738.1[医药卫生—肿瘤]