半相合基因骨髓细胞移植联合抗原免疫对小鼠肾癌的抑制作用  

Supression of tumor growth in renal cancer treatment with tumor vaccination after haploidentical bone marrow cell reconstitution

作　　者：王希友[1] 魏志涛[1] 黄建华[2] 洪宝发[1] 陈新静[2] 吕海燕[2] 杨勇[1] 

机构地区：[1]解放军总医院泌尿外科,北京100853 [2]解放军总医院免疫学教研室,北京100853

出　　处：《中华外科杂志》2010年第17期1328-1331,共4页Chinese Journal of Surgery

基　　金：全军十一五课题面上资助项目（2006266243）

摘　　要：目的 探讨半相合基因骨髓细胞移植联合抗原免疫对移植物抗肿瘤效应的影响及其机制.方法 26只BALB/c青春期小鼠为受者,随机分为单纯骨髓细胞移植组（A组,10只）、联合抗原免疫移植组（B组,10只）、对照组（C组,6只）.取BALB/c（H-2d）和C57BL/6（H-2b）交配后产生的F1代小鼠为供者.A、B组小鼠均行直线加速器全身照射（8 Gy）,照射4～6 h内经尾静脉分别注射入供者小鼠的骨髓细胞（5×106/只）,C组无处理.1周后3组小鼠均给予Renca肾癌细胞腋窝下注射（2.6×106/只）,B组小鼠在荷瘤1 d后给予33 Gy照射的Renca肾癌细胞（5×105/只）皮下注射免疫,间隔1周,重复免疫1次.通过流式细胞术（FACS）测定移植后不同时间的嵌合体水平.断颈处死小鼠后,观察所有组别受者小鼠的肿瘤质量、体积增加量、肿瘤抑瘤率,取肿瘤组织做成石蜡切片HE染色后,观察肿瘤坏死面积比,采用原位末端标记法（TUNEL）检测各组小鼠肿瘤组织的凋亡指数（AI）.结果 （1）FACS检测嵌合体水平：A、B组小鼠移植后14 d供体细胞嵌合体水平均＞90%,28 d后仍＞90%.（2）与C组和A组相比,B组的肿瘤生长速率明显减慢,肿瘤质量明显减小（P＜0.05）,体积增加量明显降低（P＜0.01）,抑瘤率高达60%,肿瘤坏死面积比明显增高（P＜0.05）,AI明显增高（P＜0.01）.结论 抗原免疫能增强半相合基因骨髓细胞移植的抗肿瘤作用,其机制可能是促进肿瘤细胞坏死和细胞凋亡增加.Objective To investigate whether whole tumor cell vaccination strategies in combination with bone marrow transplantation （BMT） can stimulate graft-versus-tumor effect （GVT）. Methods Twentysix BALB/c mice were randomly divided into 3 groups： BMT group （ group A,n = 10） ,BMT ＋ vaccination group （group B, n = 10）, control group （group C, n=6）. （BALB/c × C57BI/6） F1 mice [CB6F1, H-2K（b/d） ] were used as donors. BALB/c mice of group C were only inoculated with Renca cell （2. 6 × 106）.Mice of group A and B were conditioned with 8 Gy irridation, followed by infusion by bone marrow cell of CB6F1 mice on day 1 ,then inoculated with Renca cell （2. 6 × 106） on day 8. All mice of group B were immunized subcutaneous on the back with 5 × 105 irradiated Renca tumor cells on day 9 and day 16. All mice of group C were inoculated with Renca cell （2. 6 × 106） on day 8. In group A and B, all mice were analyzed by fluorescence activated cell sorter （FACS） on day 14, and 28 day after BMT. Mice were killed on day 32after inoculation with tumor cell and collected blood sample. All tumors were taken out to be weighed and then fixed in 10% buffered formalin, embedded in paraffin, and cut into 5 μm slices. The slices were stained with HE and examined by TdT mediated-dUTP nick end labeling （TUNEL）. Liver, skin, intestine ,and spleen were biopsied for histopathological examination. Results The results of chimera showed that engraftments of group A,B were full donor chimerism, and the chimerism of those remained above 90% and preserved even after 28 days. The tumor weight, tumor volume increment in the group B was lower than group A and C（P 〈0. 05）. The tumor suppressing rates of the group A and B were 54%, 60% respectively. The area ratio of tumor necrosis and apoptosis index （AI） of the tumor in the group B were higher than group A and C（P 〈0. 05 ）. Graft-versus-host disease was not observed in each group. Conclusion The mechanism of GVTafter haploidentical allogeneic

关 键 词：骨髓移植 异种移植模型抗肿瘤试验 移植嵌合体 细胞凋亡 

分 类 号：R737.11[医药卫生—肿瘤]