GFP Reporter Cell Line-CT26

Currently, oncolytic viruses are rapidly developing in multiple laboratories around the world, and 47 clinical trials are recruiting. Many oncolytic viruses combine targeted cytotoxicity to cancer cells with pro-inflammatory cell lysis and are also called oncolytic virus vaccines because of their potential to express immunomodulatory transgenes. Transgenic mouse cell lines were infected with EnAd expressing the GFP transgene under replication-independent or -dependent promoters. Viral mRNA expression, genome replication, and late protein expression were measured by qRT-PCR, qPCR, and Western blotting, respectively. The researchers also used Balb/c immunocompetent mice to determine the tumor-forming capacity and infectivity of the transgenic mouse cell lines. The results showed that human cancer cells can support EnAd replication, but mouse cancer cells cannot. Although mouse cells can mediate uptake of EnAd particles by expressing surface human CD46 receptors and express CMV promoter-driven transgenes, however, although E1A mRNA expression levels in mouse cells are similar to human cells, adenovirus E2B and Fibre The level of mRNA expression is blocked and less viral genome is produced. Overall, these results indicate significant differences in the early stages of replication of group C and B adenoviruses in mouse cells.

Figure 1. The researchers infected NMuMG-CD46, CT26-CD46 and their corresponding parental cell lines with EnAd viruses expressing GFP to evaluate whether human CD46 expression could promote group B adenovirus infection of mouse cells. Five days after infection, the proportion of GFP-positive cells was measured by flow cytometry. The results showed that both NMuMG-CD46 and CT26-CD46 cells showed GFP expression when using EnAd-CMV-GFP, indicating that the virus successfully entered the cells and reached the cell nucleus. (Lei J, et al., 2018)