复制缺陷性慢病毒介导的双自杀基因在小鼠异基因骨髓移植中的应用  

Application of pseudotyped lentivirus-mediated double suicide genes to allogeneic bone marrow transplantation in mice

作　　者：姜义荣[1] 陈学良[1] 马道新[1] 刘春生[1] 

机构地区：[1]山东大学齐鲁医院血液研究室

出　　处：《中华血液学杂志》2004年第2期82-86,共5页Chinese Journal of Hematology

基　　金：国家自然科学基金资助项目 (3 0 0 70 3 2 1)

摘　　要：目的　探讨复制缺陷性慢病毒介导的胞嘧啶脱氨酶 (cytosinedeaminase,CD)和胸苷激酶(thymidinekinase ,TK)双自杀基因在异基因骨髓移植 (allo BMT)时控制移植物抗宿主病 (GVHD)的可行性及其有效性。方法　将携带双自杀基因的复制缺陷性慢病毒通过脂质体转染入T细胞中。将经6 0　Coγ射线照射后的荷瘤小鼠分组进行骨髓移植 ,观察骨髓移植后荷瘤鼠CFU S、CFU GM、Y染色体整合、T细胞中CD +TK基因整合、生存率和生存期、体重及病理组织学等指标。结果　复制缺陷性慢病毒载体可将双自杀基因高效稳定转染入T细胞。在allo BMT中 ,除空白对照组外 ,各组小鼠的CFU S的数目和CFU GM产率差异无显著性 ,且均有Y染色体整合。使用前体药物治疗的小鼠未发生明显的GVHD ,其生存率和生存期明显高于其它各组 ;双自杀基因的效果优于单自杀基因。结论　复制缺陷性慢病毒介导的双自杀基因系统可在不影响移植物存活的情况下有效控制GVHD的发生 ,显著提高allo BMT的成功率。Objective To explore the feasibility and efficiency of cytosine deaminase (CD)/thymidine kinase (TK) gene-modified donor T cells used in allogeneic bone marrow transplantation (allo-BMT) as an approach to mitigate GVHD without compromising engraftment. Methods The pseudotyped lentivirus vectors containing CD and TK double suicide genes were transfected with lipofectine to donor T cells. Lethally irradiated 615 leukemia mice were transplanted with BALB/c bone marrow plus CD+TK+T cells. GVHD prophylaxis was by administration of ganciclovir (GCV) and 5-Fluoride cytosine (5-FC). Results The pseudotyped lentivirus-mediated gene transfer system could efficiently transfer CD and TK double suicide genes into donor T cells. Administration of GCV and 5-FC to the mice could markedly potentiate the CFU-S and CFU-GM yields and raise the number of peripheral white blood cells. 1×107 CD+TK+ allogeneic T cells caused GVHD of a similar magnitude and time course to that of fresh, naive T cells after allo-BMT. Administration of GCV and 5-FC in mice received CD+TK+T cells reduced the severity of GVHD and resulted in significantly longer survival as compared with non-administration mice, and the effect was stronger than that of administration of GCV or 5-FC alone. Conclusion Administration of CD+TK gene-modified donor T cells to recipient in allo-BMT might be an approach to mitigate GVHD without compromising alloengraftment.

关 键 词：复制缺陷性慢病毒 自杀基因 小鼠 异基因骨髓移植 脂质体 

分 类 号：R457.7[医药卫生—治疗学]