Immunity and Infection, University College London, London, UK.
J Gene Med. 2011 Mar;13(3):181-7. doi: 10.1002/jgm.1553.
Lentiviral vectors (LV) are promising vaccines because they transduce dendritic cells (DC) in vivo. To translate LV vaccines into clinical trials, bulk production will be necessary. The present study aimed to find a suitable envelope for LV vaccine production from stable packaging cells because the commonly used vesicular stomatitis virus envelope (VSV-G) is cytotoxic.
The envelope from Ross river virus (RRV) was selected. It can infect mouse and human cells, allowing testing in animals before clinical translation. We used VSV-G for comparison. Vectors produced with each envelope were titred on human 293T cells and mouse 3T3 cells.
RRV-pseudotyped LV (RRV-LV) infected mouse myeloid DC in culture and immunized mice. An approximately 50-fold higher dose of RRV-LV than VSV-G-LV was required to generate a similar T cell response. The RRV-LV could also be used to infect human mDC and to prime a human T cell immune response.
RRV envelope is a suitable candidate to be used for the construction of an LV producer cell line. LV vaccines with RRV envelope can be tested in mice and in human immune cell cultures. The higher dose of RRV-LV required for vaccine efficacy compared to VSV-G-LV will partly be offset by ease of production.
慢病毒载体（LV）是很有前途的疫苗，因为它们可以在体内转导树突状细胞（DC）。为了将 LV 疫苗转化为临床试验，有必要进行批量生产。本研究旨在从稳定包装细胞中寻找合适的包膜用于 LV 疫苗生产，因为常用的水疱性口炎病毒包膜（VSV-G）具有细胞毒性。
选择罗斯河病毒（RRV）的包膜。它可以感染鼠和人细胞，允许在临床转化前在动物中进行测试。我们使用 VSV-G 进行比较。用每种包膜生产的载体在人 293T 细胞和鼠 3T3 细胞上滴定。
RRV 假型 LV（RRV-LV）感染培养中的鼠髓样 DC 并免疫小鼠。与 VSV-G-LV 相比，需要大约 50 倍高剂量的 RRV-LV 才能产生相似的 T 细胞反应。RRV-LV 也可用于感染人 mDC 并引发人 T 细胞免疫反应。
RRV 包膜是构建 LV 生产细胞系的合适候选物。带有 RRV 包膜的 LV 疫苗可在小鼠和人免疫细胞培养物中进行测试。与 VSV-G-LV 相比，RRV-LV 疫苗的有效剂量较高，但生产更容易，部分抵消了这一问题。
