Poster Presentation 12th Australasian Virology Society Meeting 2024

Innovation in Enteric Virus Research: Cell Lines and in vivo shedding models to study Adenovirus F-40/41   (#242)

Valeria M Sanders 1 , Jillian M Carr 1
  1. Flinders University, ., SA, Australia

Enteric virus Adenovirus F-40/41 (AdV F-40/41) causes gastroenteritis in humans but is difficult to study due to poor replication in cell lines and challenges with long-term cultivation [1]. Primary intestinal epithelial cells can cultivate AdV F-40/41, but this method is laborious and expensive [2]. Additionally, laboratory mice are not susceptible to AdV F-40/41 infection [1]. Thus, developing standardised tools for producing high-quality viral stocks, creating cell infection models reflecting the gastrointestinal tract, and finding ways to utilize mice for studying AdV F-40/41 is crucial.

This study aims to identify permissive enteric cell lines for efficient and stable propagation of AdV F-40/41 and to develop an in vivo mouse model for studying viral shedding. We selected HuTu 80 (duodenal), PLC/PRF/5 (liver), AGS (gastric), and QGP-1 (pancreatic enterochromaffin-like) cells based on their reported susceptibility to AdV infection [3-5].

Cells were infected with a clinical isolate of AdV F-40/41, and viral replication kinetics were assessed by PCR four days post-infection. Results showed all tested cell lines were permissive to AdV F-40/41. Serial passage maintained infectivity in AGS and PLC/PRF/5 cells after four passages, whereas HuTu 80 cells did not sustain viral replication beyond the first passage. The QGP-1 cell line is yet to be fully tested, including studies using spheroids generated from this cell line.

In a mouse model, 4-week-old mice were orally challenged with AdV F-40/41, and viral shedding in faecal pellets was analysed by PCR and in vitro infectivity. Results demonstrated rapid viral shedding in feces from 2-7 hours post-challenge, declining to low levels at 24 hours.

We have successfully developed models using multiple enteric-related cell lines to study AdV F-40/41 replication and a mouse model for viral shedding. These models are useful for testing novel compounds for virus attenuation and provide standardized, reproducible methods for cultivating AdV F-40/41.

 

  1. Kosulin K. 2019. Intestinal HAdV Infection: Tissue Specificity, Persistence, and Implications for Antiviral Therapy. Viruses 11:804.
  2. Holly MK, Smith JG. 2018. Adenovirus Infection of Human Enteroids Reveals Interferon Sensitivity and Preferential Infection of Goblet Cells. J Virol 92.
  3. Becker M, Conca DV, Dorma N, Mistry N, Hahlin E, Frängsmyr L, Bally M, Arnberg N, Gerold G. 2023. Efficient clathrin-mediated entry of enteric adenoviruses in human duodenal cells. Journal of Virology 97:e00770-23.
  4. Grabow WOK, Puttergill DL, Bosch A. 1992. Propagation of adenovirus types 40 and 41 in the PLC/PRF/5 primary liver carcinoma cell line. Journal of Virological Methods 37:201-207.
  5. Westerberg S, Hagbom M, Rajan A, Loitto V, Persson BD, Allard A, Nordgren J, Sharma S, Magnusson KE, Arnberg N, Svensson L. 2018. Interaction of Human Enterochromaffin Cells with Human Enteric Adenovirus 41 Leads to Serotonin Release and Subsequent Activation of Enteric Glia Cells. J Virol 92.