人IL-12和B7-1基因对HuPBL-SCID小鼠模型中人源肿瘤的协同抑制作用  

Establishment of a SCID Mouse Model for Synergistic Anti-tumor Effect of Human IL-12 and B 7-1

作　　者：高军[1] 赵平[2] 陈秀华 李茂[1] 龙建秋[1] 谷爱梅[1] 吴晓兰[1] 曹广文[2] 戚中田[2] 

机构地区：[1]海军医学研究所流行病学研究室,上海200433 [2]第二军医大学微生物学教研室,上海200433 [3]上海市医药工业研究院药理研究室,上海200040

出　　处：《生物化学与生物物理学报》2003年第6期529-535,共7页

基　　金：国家自然科学基金资助项目 (No .3 960 0 172 ;No .3 0 1710 5 5 )~~

摘　　要：人IL 12 (hIL 12 )对鼠源的免疫细胞作用较弱 ,进行hIL 12抗瘤研究缺乏有效的动物模型。为此利用荷人瘤的HuPBL SCID小鼠模型 ,将改良的Winnassay细胞免疫功能分析方法应用于评价hIL 12和hB 1协同抗瘤作用。用商业化的非脂质转基因试剂配制单独的hIL 12、hB7 1或hIL 12联合hB7 1的基因转染液。将上述三种基因转染液分别瞬时转基因入人恶性黑色素瘤A375细胞 ,转基因 30min后 ,按组别 (hIL 12组、hB7 1组、hIL 12 +hB7 1组、rhIL 2对照组、HuPBL对照组和肿瘤对照组 )同人外周血淋巴细胞混合接种于SCID小鼠皮下 ,2 0天后处死实验小鼠 ,测得hIL 12和hB7 1的抑瘤率分别为 74 .0 6 %和 6 6 .98% ,联合作用达 93.4 0 % ,rhIL 2和HuPBL对照组无抗瘤作用。同时 ,在此模型上hIL 12联合hB7 1基因对人大肠癌LoVo和人肺癌SPC的抑制率分别为 98.37%和 97.39%。实验建立的各组HuPBL SCID小鼠 2 0天的外周血都含有人IgG(>0 .5mg/L)和极少量人CD3+ 淋巴细胞 (>0 .5个 /10 0个有核细胞 )。hIL 12、hB7 1和hIL 12 +hB7 1组的瘤灶内人淋巴细胞浸润生长 ,肿瘤细胞残存 ,而rhIL 2和HuPBL对照组的瘤灶内人淋巴细胞“似团样”生长 ,肿瘤生长旺盛。表明此HuPBL SCID小鼠模型上的抗瘤机制为局部免疫反应。Human IL-12(hIL-12) has weak effect on mouse immunity cells, so the practical animal model is not available for the study of hIL-12 anti-tumor activity. In this work, the improved Winn assay was applied to evaluate the synergistic anti-tumor effects of hIL-12 and human costimulatory molecule B7-1(hB7-1) on human tumor in HuPBL-SCID mouse model. Three gene transferring solutions? ? WTBX]hIL -12, hIL -12, hB7 -1 and their mixture(1∶1) —were prepared using the nonliposome transgene reagent and the expressing vectors, and hIL -12, hB7 -1 or their mixture were transferred into tumor cell A375 respectively. Then A375 were co-injected into SCID mice with HuPBL, and rhIL-2 were injected i.p. as an anti-tumor agitator. On the other hand, LoVo and SPC tumor cells were also used to test the inhibitory effect of the mixture of hIL -12 and hB7 -1. The anti-tumor effect of transferred genes was estimated by detecting tumor inhibition rate. Furthermore, the histochemical change of A375 implanting tumor tissue was also observed. Results showed that, to A375, the tumor inhibition rate of hIL -12, hB 7-1, or their mixture were 74.06%, 66.98%, and 93.40%, respectively ( P <0.01); and the mixture showed a good synergistic effect according to the Webb’s fraction multiplication law. The tumor inhibition rate of the mixture in LoVo and SPC implanted mice were 98.37% and 97.39% respectively, also showing a good synergistic effect. Histochemical study in A375 implanted mice showed that in gene transfected mice, tumor cells were greatly inhibited and fully intruded by HuPBL cells; while in control group, tumor cells grew very well and HuPBL showed a conglomeration. At last, the human IgG and T cells in PBL of HuPBL-SCID mice were higher than non-HuPBL-SCID mice implanted A375; which showed that HuPBL-SCID mice could be applied for the evaluation of the anti-tumor effect of human IL-12 and B7-1. All data indicated that the combination of hIL -12 and hB 7-1 gene might be a promising approach for in

关 键 词：肿瘤 HuPBL-SCID小鼠 基因治疗 B7-1基因 IL-12基因 

分 类 号：R730.3[医药卫生—肿瘤]