HSV—tk／NAS系统基因治疗脑肿瘤中野生型腺病毒的检测及其安全性

在腺病毒介导的ＨＳＶ－ｔｋ／ＮＡＳ系统脑肿瘤基因治疗研究过程中，建立了野生型腺病毒（ＲＣＡ）的检测系统，主要包括腺病毒Ｅ１区的ＰＣＲ以及野生型病毒的细胞病理效应观察这两种方法，ＰＣＲ的灵敏度为１个２９３细胞所含有的Ｅ１片段数。对基因治疗脑肿瘤所采用的重组腺病毒ＡｄＴＫ及其感染的细胞进行ＲＣＡ的检测，没有发现ＲＣＡ。通过离体实验首次发现，重组腺病毒对脑肿瘤细胞的杀伤作用明显高于正常的脑胶质细胞；大鼠

REPLICATION-COMPETENT ADENOVIRUS DETECTION IN HSV-tk/NAS GENE THERAPY FOR BRAIN TUMOR AND ITS SAFETY

Based on HSV-tk/NAS gene therapy for brain tumor,the system for detection of replication-competent adenovirus(RCA) was studied.It included PCR for E1 region of adenovirus and cytopathological effect observation.The sensitivity of PCR is denoted by the copies of E1 region in one 293 cell.When the recombinant adenovirus AdTK and the affected cells were detected for RCA,all the results were negative.In in vitro study,C6 glioma cells were more sensitive to GCV than the primary astrocytes;rats and rabbits received intracerebral injection of AdTK with GCV resulted moderate necrosis and inflammation in normal brain tissue.AdTK were detected by PCR in brain at most 4 weeks.Mice and rats injected with AdTK via tail vein with GCV,resulted mild inflammatory response only in lung and liver.