转染sFas基因骨髓移植阻抑白血病小鼠肿瘤细胞的免疫逃逸  

Blocking tumor cell immune escape of leukemic mouse in bone marrow transplantation by transfection of soluble Fas gene

作　　者：黎纬明[1] 赵智刚[1] 邹萍[1] 胡中波[1] 李爱香[1] 刘凌波[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液科湖北省血液病研究所,武汉430022

出　　处：《中华器官移植杂志》2006年第7期403-406,共4页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金资助(No.39970708)

摘　　要：目的探讨骨髓细胞转染小鼠可溶性Fas（sFas）基因后移植能否阻抑白血病小鼠肿瘤细胞的免疫逃逸。方法用AdEasy系统构建携带sFas和EGFP的腺病毒质粒，先按病毒／细胞的感染倍数（MOI）为100的比例分别直接感染呈对数生长期的EL4细胞，培养24h后，采用氚标记胸腺嘧啶掺入法检测其诱导YAC-1细胞凋亡的情况。然后同法以携带sFas和EGFP的腺病毒质粒分别转染雄性C57BL／6小鼠骨髓细胞，每只雌性C57BL／6小鼠（白血病模型）通过尾静脉接受5×10^6～10×10^6个转染sFas或EGFP的雄性C57BL／6小鼠骨髓单个核细胞移植。移植后观察受者的造血重建情况、白血病／淋巴瘤的发生及生存率。结果体外实验中，转染EGFP的EL4细胞诱导YAC-1细胞凋亡的情况与非转染的EL4细胞相似，转染sFas基因后，YAC-1细胞的凋亡率明显下降（P〈0．01）。接受转染sFas基因的骨髓移植者，移植后8周受者全部存活（16／16），骨髓中未发现EL4细胞，造血功能恢复良好，能检测到Y染色体；接受转染EGFP的骨髓移植者，移植后8周仅1只小鼠（1／16）存活（P〈0．01），在死亡和存活的小鼠中均存在EL4细胞，其造血重建情况较转sFas者差。结论转染sFas基因的造血细胞移植可以在体内阻抑肿瘤细胞的免疫逃逸，同时不影响造血重建功能。Objective To study whether transfection of soluble Fas gene could block tumor cells＇ immune escape in morrow marrow transplantation （BMT）. Methods The two recombinant adenoviral vectors containing soluble Fas and EGFP were constructed with the AdEasy adenovirus vector system. The two vectors were transfected into EL4 cell with MOI of 100. Twenty-four h later, the apoptosis ratio of the target cells YAC-1 induced by EL4 cells transfected with sFas or EGFP was detected by 3 H thymidine （3 H-TdR） labeling. The two recombinant adenoviral vectors containing soluble Fas and EGFP were transfected into BM cells of C57BL/6 mouse in the same way. Then every remale leukemic C57BL/6 mouse received 5 × 10^6-10×10^6 bone marrow mononuclear cells of male C57BL/6 mouse that transfected with sFas and EGFP vector. The hematopoietic reconstitution, generation of leukemia/lymphoma and the survival rate were observed in all the groups after BMT. Results The apoptosis of YAC-1 cells was similar while induced by EGFP transfected or non-transfected EL4 cells, and transfection of sFas could decrease the apoptosis of YAC-1 cells in vitro （P〈0. 01）. Moreover, mice transplanted with sFas transfected bone marrow cells survived 8 weeks post-transplantation （16/16）, no EL4 cells were seen in bone marrow, hematopoiesis recovered soon, and the Y chromatosomes were detectable. While the mice were transplanted with EGFP transfected bone marrow cells, only 1 out of 16 survived （P〈0. 01）. EL4 cells could be found in either dead or survival mice, and hematopoiesis recovery was not good. Conclusion The immune escape of tumor cells was blocked by transplantation of hematopoietic cells transfected with sFas gene in vivo, and the ability of hematopoietic reconstruction of hematopoeitic cells was not affected by transfection of sFas gene.

关 键 词：转染 肿瘤逃逸 骨髓移植 白血病 

分 类 号：R733.7[医药卫生—肿瘤]