PGK驱动逆转录病毒载体介导HSV-TK／GCV控制GVHD的研究  

The experimental study of preventing and treating GVHD mediated by the retroviral vectors carrying the herpes simplex virus-thymidine kinase/gancidovir（HSV-TK/GCV）under PGK promoter

作　　者：朱锋[1] 徐开林[1] 杜冰[1] 陈香梅[1] 潘秀英[1] 

机构地区：[1]江苏省徐州医学院附属医院血液科,徐州221102

出　　处：《中华微生物学和免疫学杂志》2007年第4期306-310,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（30170389）;江苏省“135”医学重点人才项目（RC2002080）;江苏省高校自然科学研究计划（02KJB320013）

摘　　要：目的：研究磷酸甘油酸激酶（PGK）启动子驱动的逆转录病毒载体介导单纯疱疹病毒胸苷激酶/更昔洛韦（HSV-TK／GCV）系统在小鼠异基因骨髓移植（allo-BMT）后能否减轻移植物抗宿主病（GVHD）。方法：将转HSV-TK基因的逆转录病毒感染供鼠（C57BL/6鼠）脾淋巴细胞，感染后细胞与供鼠骨髓细胞混合移植给Coγ7射线照射后的受鼠（BALB／c鼠）。移植后腹腔注射GCV，用药1周，按移植后首次给药时间，分为GCV0d组、GCV7d组、GCV12d组。观察移植后受鼠生存期、存活率、GVHD发生率及严重程度、T细胞免疫重建（CD3、CD4、CD8）及异基因嵌合等指标。结果：TK／GCV0d组、TK／GCV7d组小鼠平均生存时间分别为（26.90±4.88）d、（27.00±4.80）d，较TK／GCV12d组（21.504±3.26）d和移植对照组（15.10±O.43）d明显延长（P〈0.05），TK／GCV0d组与TK／GCV7d组之间比较差异无统计学意义（P〉0.05）。对照组小鼠12～14d100％发生Ⅲ～Ⅳ级GVHD，实验组死亡小鼠有Ⅱ～Ⅳ级GVHD病理学改变，而长期生存小鼠仅出现Ⅰ～Ⅱ级GVHD。TK／GCV0d组、TK/Gcv7d组、TK／GCV12d组在移植后5、10、15d3个时间点检测CD4^＋ T细胞均高于对照组（P〈0.05），CD8^＋ T细胞均低于对照组（P〈0.05）。移植后30d检测10只受鼠异基因嵌合率均在95％～100％。结论：PGK启动子驱动的逆转录病毒载体介导HSV-TK／GCV系统能有效控制小鼠allo-BMT后GVHD，移植后早期预防性用药效果优于发生GVHD后治疗性用药。Objective To study whether herpes simplex virus-thymidine kinase/ganciclovir（HSV-TK/ GCN） induced by the retroviral vectors. Metlmds Donor splenic lymphocytes were infected by retroviral vectors carrying HSV-TK. With donor bone marrow cells, they were both transplanted into recipient mice irradiated by ^60 Co γ ray. GCV was administered for 7 days by intraperitoneal injection after transplantation. The surviving time, incidence of GVHD, T lymphocytes immunity reconstruction and the ratio of allogeneic chimeras were detected after allo-BMT. Results The average survival time of mice in GCV 0 d group and GCV 7 d group and survival rates over 50 days were both longer than GCV 12 d group and transplantation control group. The incidence of Ⅲ-Ⅳ GVHD after allo-BMT in transplantation control group was 100%.After allo-BMT, CD4^＋ T lymphncytes, CD4/CU8 of experimental group were all higher than that of the transplantation control group, but CD8^＋ T lymphocyte eOunting was less than the latter＇s. Conclusion HSV-TK/GCN induced by the retroviral vectors,which were driven by PGK promoter had a certain effect on prevention and treat of GVHD. The effect of prophylactic administration in early period after transplantation was better than therapeutic administration when GVHD had occurred.

关 键 词：逆转录病毒载体 HSV-TK/GCV 移植物抗宿主病 异基因移植 

分 类 号：R686[医药卫生—骨科学]