# Development of a Porcine Cell Line Stably Expressing Ephrin‐B2 for Nipah Virus Research and Diagnostic Testing

## TL;DR

Researchers developed a pig cell line that stably expresses ephrin-B2, improving Nipah virus research and diagnostic testing.

## Contribution

A novel pig-derived cell line stably expressing ephrin-B2 is developed for enhanced Nipah virus propagation and diagnostics.

## Key findings

- PK-15/Ephrin-B2 cells showed over 1000-fold increased NiV pseudovirus infectivity compared to wild-type cells.
- These cells exhibited higher virus susceptibility than Vero cells by more than 30-fold.
- Stat2-knockout PK-15/Ephrin-B2 cells maintained stable infectivity in the presence of type I interferon.

## Abstract

Nipah virus (NiV) is a highly pathogenic zoonotic virus transmitted from bats to humans through pigs as a crucial intermediate host. NiV outbreaks pose significant public health and economic threats, especially for pig farmers. Although the World Organization for Animal Health recommends African green monkey‐derived Vero cells for NiV isolation, pig‐derived cell lines could represent an optimal platform for propagating pig‐origin NiV, as signs and symptoms of NiV infection differ among different hosts. In this study, we generated and evaluated pig‐derived PK‐15 cells stably expressing pig‐derived ephrin‐B2 (PK‐15/Ephrin‐B2 cells), the primary receptor for NiV. NiV pseudovirus infectivity was increased by > 1000‐fold in PK‐15/Ephrin‐B2 cells compared with that in wild‐type PK‐15 cells, whereas virus susceptibility was higher in PK‐15/Ephrin‐B2 cells than in Vero cells (> 30‐fold). Furthermore, Stat2‐knockout PK‐15/Ephrin‐B2 cells exhibited stable viral infectivity in the presence of type I interferon, making it particularly suitable for clinical sample testing. Moreover, PK‐15/Ephrin‐B2 cells proved useful for neutralization tests using anti‐NiV hyperimmune ascitic fluid. Therefore, PK‐15 cells expressing pig ephrin‐B2 could represent an efficient tool for virus isolation, vaccine development, and virological studies of NiV and related henipaviruses.