A polarized cell system amenable to subcellular resolution imaging of influenza virus infection
Naffakh, N.; Brault, J.-B.; Thouvenot, C.; Cannata-Serio, M.; Paisant, S.; Fernandez, J.; Geny, D.; Danglot, L.; Mallet, A.
Show abstract
The life cycle of influenza A viruses (IAV), and notably intracellular trafficking of the viral genome, depend on multiple interactions with the cellular cytoskeleton and endomembrane system. A limitation of the conventional cellular models used for mechanistic study and subcellular imaging of IAV infection is that they are cultured in two dimensions (2D) under non-polarizing conditions, and therefore they do not recapitulate the intracellular organization of the polarized respiratory epithelial cells naturally targeted by IAVs. To overcome this limitation, we developed an IAV-infection assay in a 3D cell culture system which allows imaging along the baso-lateral axis of polarized cells, with subcellular resolution. Here we describe a protocol to grow polarized monolayers of Caco2-TC7 cells on static Cytodex-3 microcarrier beads, infect them with IAV, and subsequently perform immunostaining and confocal imaging, or electron microscopy, on polarized IAV-infected cells. This method can be extended to other pathogens that infect human polarized epithelial cells.
Matching journals
The top 6 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Efficient clathrin-mediated entry of enteric adenoviruses in human duodenal cells 96%
- Macropinocytosis and Clathrin-Dependent Endocytosis Play Pivotal Roles for the Infectious Entry of Puumala Virus. 96%
- Birnaviridae virus factories show features of liquid-liquid phase separation, and are distinct from paracrystalline arrays of virions observed by electron microscopy 95%
Similar papers in this journal
- The replication of the human respiratory syncytial virus in a T cell line has multiple ineffective steps 95%
- Coronavirus RNA synthesis takes place within membrane-bound sites 95%
- Single-cell virology: On-chip, quantitative characterization of the dynamics of virus spread from one single cell to another 94%
Similar papers in this journal
Similar papers in this journal
- Human parvovirus B19 interacts with globoside under acidic conditions as an essential step in endocytic trafficking 95%
- Analysis of a fully infectious bio-orthogonally modified human virus reveals novel features of virus cell entry 94%
- Rhinovirus C replication is associated with the endoplasmic reticulum and triggers cytopathic effects in an in vitro model of human airway epithelium 94%
Similar papers in this journal
- Zika virus hijacks extracellular vesicle tetraspanin pathways for cell-to-cell transmission 93%
- Sustained replication of synthetic canine distemper virus defective genomes in vitro and in vivo 92%
- Inactivation mechanisms of Influenza A virus under pH conditions encountered in aerosol particles as revealed by whole-virus HDX-MS. 91%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.