STUDY OF A MURINE RETROVIRUS FROM A PACKAGING CELL LINE USED IN GENE THERAPY

可复制的鼠逆转录病毒（RCR）可以在一些细胞中产生。 用于制备基因治疗产品的包装细胞系的病例 供人类使用。 由于RCR在人体中的相关风险是 建议对RCR进行未知的严格测试，以排除可能的 产品污染。 以前的研究中，正常和 中度免疫抑制的猴子接种RCR表明， 病毒清除和无病理学（Cornetta等，1991）;然而，最近 研究表明，RCR与淋巴瘤的发展有关， 严重免疫抑制的恒河猴（Donahue et al，1992）。 评估 RCRs在人类中的潜在风险，我们着手分析RCRs的作用 在感染和淋巴瘤发生中使用猴子作为灵长类模型系统。 四只正常幼年恒河猴接种RCR制剂 购自Genetic Therapy Inc. 目前正在进行研究， 猴感染性使用血液样本获得每两个星期后， 接种：检测PBL的病毒分离;获得血浆， 通过Western印迹分析进行抗体检测;从PBLs制备DNA， 通过PCR分析病毒DNA。此外，临床血液学和血清 化学正在进行。

Replication-competent murine retroviruses (RCRs) can be generated in some cases from packaging cell lines used in preparation of gene therapy product generated for human use. Since the risk associated with RCR in man is unknown, rigorous testing for RCRs is recommended to exclude possible contamination of the product. Previous studies in which normal and moderately immunosuppressed monkeys were inoculated with RCR indicated rapi virus clearance and no pathology (Cornetta et al, 1991); however, a recent study has shown RCR to be associated with development of lymphomas in severely immunosuppressed rhesus monkeys (Donahue et al, 1992). To assess the potential risk of RCRs in man, we undertook to analyze the role of RCRs in infection and lymphomagenesis using monkeys as a primate model system. Four normal juvenile rhesus monkeys were inoculated with a RCR preparation obtained from Genetic Therapy Inc. Studies are in progress to analyze the monkeys for infectivity using blood samples obtained every two weeks post- inoculation: PBLs are tested for virus isolation; plasma is obtained for antibody detection by Western blot analysis; DNAs are prepared from PBLs fo analysis of viral DNA by PCR. In addition clinical hematology and serum chemistry is being performed.