激活骨髓抗肿瘤生物效应的实验研究  

An experimental study on the anti tumor biotherapy of activated bone marrow

作　　者：刘子玲[1] 王冠军[1] 易永林[1] 姚程[1] 张福明[1] 

机构地区：[1]白求恩医科大学第一临床学院血液内科

出　　处：《中华内科杂志》1998年第10期659-662,共4页Chinese Journal of Internal Medicine

摘　　要：目的最大限度的增加激活骨髓（ＡＢＭ）的数量和抗肿瘤活性，保持ＡＢＭ的重建造血功能，探讨ＡＢＭ的性质和杀伤肿瘤细胞的机制。方法应用植物血凝素（ＰＨＡ）、抗ＣＤ３单抗、重组白细胞介素２（ｒＩＬ２）联合作用体外激活骨髓，采用甲基纤维素半固体培养法进行粒巨噬细胞系祖细胞（ＣＦＵＧＭ）、红系祖细胞（ＢＦＵＥ）培养，以ＭＴＴ比色分析法测定ＡＢＭ的杀伤活性，应用流式细胞仪检测ＡＢＭ细胞表面ＣＤ２５、ＦａｓＬｉｇａｎｄ（ＦａｓＬ）表达。结果ｒＩＬ２诱导的ＡＢＭ体外培养１天即可诱导出杀伤活性，第５天最强，发病期、缓解期和正常对照组ＡＢＭ的杀伤活性（％）分别为３１．１±４．０、５４．２±４．０、５８９±６．７，缓解期与正常对照组差异无显著性（Ｐ＞０．０５）。ｒＩＬ２、抗ＣＤ３单抗＋ｒＩＬ２、ＰＨＡ＋抗ＣＤ３单抗＋ｒＩＬ２体外诱导３天的ＡＢＭ杀伤活性（％）分别为５６．４±９．０、６５．８±９．２、７２．５±８．９，第７天细胞增殖倍数分别为３．７、６．０、１０．０倍。体外激活３～５天的ＡＢＭ与同期培养的未激活骨髓细胞相比ＣＦＵＧＭ、ＢＦＵＥ形成率差异无显著性（Ｐ＞０．０５）。结论合理的多种细胞因子联合应用?Objective To increase maximally the amount and anti tumor activity of activated bone marrow (ABM), so as to keep the hematopoietic reconstruction function of ABM and to study the proper of ABM and its mechanism of killing tumor cells. Methods At first, using PHA, anti CD 3 monoclonal antibody and rIL 2 to activate jointly bone marrow outside the body, then cultivating CFU GM and BFU E with methylcellulose semi solid culture method and at last testing the killing activity of ABM with MTT method and monitoring the content of CD 25 and Fas Ligand (FasL) on the surface of ABM cells with flowing cytometry. Results The killing activity of ABM induced by rIL 2 appeared on the first cultivating day and reached its maximum on the fifth day. The percentage of the killing activity of ABM induced by rIL 2 was 31.1±4.0 and 54.2±4.0 respectively in acute stage and remission period of leukemic patients. The latter percentage approached that of a normal control group ( P >0.05), in which killing activity of marrow, was 58.9±6.7. The percentage of the killing activity of ABM induced outside of the body by rIL 2, anti CD 3 monoclonal antibody+rIL 2 and PHA+anti CD 3 monoclonal antibody+rIL 2 were 56.4±9.0, 65.8±9.2 and 72.5±8.9 respectively and the cell proliferation after seven days was 3.7, 6.0 and 10.0 times respectively. Comparing the CFU GM and BFU E formation rate of ABM activated for 3～5 days outside the body with the marrow cells that were activated but cultivated for the same time, there was no obvious difference ( P >0.05). Conclusion Reasonable joint application of various cytokines can strengthen the killing activity of ABM and raise the cells proliferation ability. ABM can keep hematopoietic reconstruction function. The mechanism killing tumor cells by ABM is related not only to the participation of T cells which is a component of ABM, but also to the apoptosis of Fas/FasL medium tumor cells.

关 键 词：白细胞介素-2 CD3抗原 植物血凝素 骨髓激活 

分 类 号：R730.51[医药卫生—肿瘤]