Development and characterization of a packaging cell line for pseudo-infectious yellow fever virus particle generation

Abstract

Introduction: Pseudo-infectious yellow fever viral particles (YFV-PIVs) have been used to study vaccines and viral packaging. Here, we report the development of a packaging cell line, which expresses the YFV prM/E proteins. Methods: HEK293 cells were transfected with YFV prM/E and C (84 nt) genes to generate HEK293-YFV-PrM/E-opt. The cells were evaluated for their ability to express the heterologous proteins and to package the replicon repYFV-17D-LucIRES, generating YFV-PIVs. Results: The expression of prM/E proteins was confirmed, and the cell line trans-packaged the replicon for recovery of a reporter for the YFV-PIVs. Conclusions: HEK293-YFV-prM/E-opt trans-packaging capacity demonstrates its possible biotechnology application.

REPLICAID: replicon vaccines to combat avian infectious diseases

“…Ultimately, HEK293T cells were selected, as these cells maintained their high viral titers even in the presence of observable cytopathic effects. HEK293 cells are commonly used as a platform for the propagation or production of flavivirus VLP vaccines (Powers et al, 2022;Queiroz et al, 2018), adenovirus-based COVID-19 vaccines (Joe et al, 2022), and studying permissive to persistent flavivirus infections (Mlera et al, 2015(Mlera et al, , 2016Sempere & Arias, 2019). Contrary to earlier reports of TMUV sensitivity to human interferon (Ma et al, 2019) and the moderate susceptibility to TMUV infection reported for HEK293T cells (Ruangrung et al, 2021), our experiments demonstrate an early onset of virus propagation and high virus titers at 72 hpi.…”