Department of Surgery, University of Minnesota, Minneapolis, MN 55455, USA.
Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA.
Viruses. 2023 Jul 1;15(7):1495. doi: 10.3390/v15071495.
There have been numerous advances in gene therapy and oncolytic virotherapy in recent years, especially with respect to cutting-edge animal models to test these novel therapeutics. With all of these advances, it is important to understand the biosafety risks of testing these vectors in animals. We performed adenovirus-based viral shedding studies in murine models to ascertain when it is appropriate to downgrade the animals from Biosafety Level (BSL) 2 to BSL 1 for experimental handling and transport. We utilized intravenous injections of a replication-competent adenovirus and analyzed viral shedding via the collection of buccal and dermal swabs from each animal, in addition to obtaining urine and stool samples. The adenovirus hexon copy number was determined by qPCR, and plaque formation was analyzed to assess the biologic activity of viral particles. Our results demonstrate that after 72 h following viral inoculation, there is no significant quantity of biologically active virus shedding from the animals. This observation suggests that on day 4 following adenovirus injection, mice can be safely downgraded to BSL 1 for the remainder of the experiment with no concern for hazardous exposure to laboratory personnel.
近年来，基因治疗和溶瘤病毒治疗取得了许多进展，特别是在用于测试这些新型疗法的尖端动物模型方面。随着所有这些进展，了解在动物中测试这些载体的生物安全风险非常重要。我们在小鼠模型中进行了基于腺病毒的病毒脱落研究，以确定何时可以将动物从生物安全级别 2 (BSL2) 降级为实验处理和运输的生物安全级别 1 (BSL1)。我们通过收集每个动物的口腔和皮肤拭子，以及获得尿液和粪便样本，利用复制型腺病毒进行静脉注射，并分析病毒脱落情况。通过 qPCR 确定腺病毒六邻体拷贝数，并分析空斑形成以评估病毒颗粒的生物学活性。我们的结果表明，在病毒接种后 72 小时，动物没有明显数量的具有生物活性的病毒脱落。这一观察结果表明，在腺病毒注射后的第 4 天，小鼠可以安全地降级为 BSL1，在实验的其余时间内，实验室人员不会有危险接触的风险。
