单纯疱疹病毒胸苷激酶基因工程化细胞和更昔洛韦系统的急慢性毒性

Toxicity studies of retroviralmediated herpes simplex virus thymidine kinase gene transfer and ganciclovir administration

目的：探讨单纯疱疹病毒Ⅰ型胸苷激酶基因工程化细胞（ｐＬＴＫｃＳＮ／ＶＰＣ）和更昔洛韦（ＧＣＶ）系统的动物体内急、慢性毒性。方法：将ｐＬＴＫｃＳＮ／ＶＰＣ和（或）以β－半乳糖苷酶基因为报告基因的ＰＡ３１７细胞悬液注入小鼠、大鼠和灵长类动物体内，７ｄ后部分动物再全身应用ＧＣＶ。分批处死动物，观察ｐＬＴＫｃＳＮ／ＶＰＣ和ＧＣＶ系统在不同种属动物的体内急、慢性毒性。采用ＰＣＲ和原位杂交方法直接检测胸苷激酶基因在动物脑和全身各脏器内的整合及表达情况。结合原位杂交和Ｘ－ｇａｌ染色结果，判断注射细胞在动物脑内的存活时间。结果：ｐＬＴＫｃＳＮ／ＶＰＣ和ＧＣＶ系统对动物的心、肝、肺和肾有一定的急性损害作用。该细胞在大鼠和猴脑内的存活时间为３周。

Objective: To study the toxicities of the retroviralvector producer cells of herpes simplex virus thymidine kinase gene(pLTKcSN/VPC) and the ganciclovir(GCV) system in mice, rats and primates. Methods: pLTKcSN/VPC and/or the cell suspension of PA317 cells modified by the βgal reporter gene were injected into the animal bodies, and 7 d later, some of the animals were given GCV. The animals were then sacrificed as scheduled, and the acute and long term toxicities of this treatment system were evaluated through histological studies and direct detection of the transduction and expression of the thymidine kinase (TK) gene in animal brain and other organs. The survival interval of the xenogenic cells in rat and monkey brain was determined by the Xgal staining and the  in situ  hybridization of the TK gene in brain. Results: The pLTKcSN/VPC and GCV system had certain acute side effects on heart, liver, lung and kidney, but no long term toxicities could be detected. The survival time of pLTKcSN/VPC in the brain of rats and primates was 3 weeks. Conclusion: The pLTKcSN/VPC and GCV system has slight toxicities when applied in vivo. 