A Method for the Production of Recombinant VSVs with Confirmation of Biological Activity.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-01-01 DOI:10.32607/actanaturae.27314

V D Moroz, N B Gasanov, A D Egorov, A S Malogolovkin, M O Nagornykh, E N Subcheva, E S Kolosova, A Yu Fizikova, R A Ivanov, A V Karabelsky

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Abstract

The design of new effective cancer treatment methods is a promising and important research field in translational medicine. Oncolytic viruses can induce immunogenic cell death by activating the body's immune system to recognize tumor cells. This work presents the results for optimizing the production of recombinant vesicular stomatitis viruses (rVSVs). To ensure the assembly of viral particles, we developed the HEK293TN-T7 cell line, which stably expresses DNA-dependent RNA polymerase 7 for viral genome transcription, and obtained helper plasmids encoding viral genes under the control of the CAG promoter. The oncolytic activity of the purified virus preparation was assessed in a murine model of B16F10Red melanoma cells expressing a red fluorescent protein. The presented method makes it possible to obtain purified viral preparations with a high titer and oncolytic activity. The amplification of viral particles in a HEK293 suspension culture allows for rapid scalability. Therefore, the developed approach can be used to obtain other recombinant VSV-based oncolytic viruses for tumor immunotherapy.

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一种生产重组 VSV 并确认其生物活性的方法。

设计新的有效癌症治疗方法是转化医学中一个前景广阔的重要研究领域。肿瘤溶解病毒可通过激活人体免疫系统识别肿瘤细胞来诱导免疫性细胞死亡。这项工作展示了优化重组水泡性口炎病毒（rVSV）生产的成果。为确保病毒颗粒的组装，我们开发了 HEK293TN-T7 细胞系，它能稳定表达用于病毒基因组转录的 DNA 依赖性 RNA 聚合酶 7，并获得了在 CAG 启动子控制下编码病毒基因的辅助质粒。在表达红色荧光蛋白的 B16F10Red 黑色素瘤小鼠模型中评估了纯化病毒制剂的溶瘤活性。所介绍的方法可以获得具有高滴度和溶瘤活性的纯化病毒制剂。在 HEK293 悬浮培养物中扩增病毒颗粒可实现快速扩展。因此，所开发的方法可用于获得其他基于 VSV 的重组溶瘤病毒，用于肿瘤免疫疗法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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