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Title: Investigating virus-host cell interactions: Comparative binding forces between hepatitis C virus-like particles and host cell receptors in 2D and 3D cell culture models.
Austin Authors: Collett, Simon;Torresi, Joseph ;Silveira, Linda Earnest;Truong, Vi Khanh;Christiansen, Dale;Tran, Bang M;Vincan, Elizabeth;Ramsland, Paul A ;Elbourne, Aaron
Affiliation: Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, VIC 3010, Australia
Department of Immunology, Monash University, Melbourne, VIC 3004, Australia
Surgery (University of Melbourne)
Department of Infectious Diseases, Melbourne Medical School, University of Melbourne, The Peter Doherty Institute for Infection and Immunity, Melbourne 3000, Australia
Victorian Infectious Diseases Reference Laboratory, The Peter Doherty Institute for Infection and Immunity, Melbourne, VIC 3000, Australia
School of Pharmacy and Biomedical Sciences, Curtin University, Perth, Western Australia 6102, Australia
School of Science, College of Science, Engineering and Health, RMIT University, Melbourne, VIC 3000, Australia
Issue Date: 15-Jun-2021
Date: 2021-02-23
Publication information: Journal of Colloid and Interface Science 2021; 592: 371-384
Abstract: Cell cultures have been successfully used to study hepatitis C virus (HCV) for many years. However, most work has been done using traditional, 2-dimensional (2D) cell cultures (cells grown as a monolayer in growth flasks or dishes). Studies have shown that when cells are grown suspended in an extra-cellular-matrix-like material, they develop into spherical, 'organoid' arrangements of cells (3D growth) that display distinct differences in morphological and functional characteristics compared to 2D cell cultures. In liver organoids, one key difference is the development of clearly differentiated apical and basolateral surfaces separated and maintained by cellular tight junctions. This phenomenon, termed polarity, is vital to normal barrier function of hepatocytes in vivo. It has also been shown that viruses, and virus-like particles, interact very differently with cells derived from 2D as compared to 3D cell cultures, bringing into question the usefulness of 2D cell cultures to study virus-host cell interactions. Here, we investigate differences in cellular architecture as a function of cell culture system, using confocal scanning laser microscopy, and determine differences in binding interactions between HCV virus-like particles (VLPs) and their cognate receptors in the different cell culture systems using atomic force microscopy (AFM). We generated organoid cultures that were polarized, as determined by localization of key apical and basolateral markers. We found that, while uptake of HCV VLPs by both 2D and 3D Huh7 cells was observed by flow cytometry, binding interactions between HCV VLPs and cells were measurable by AFM only on polarized cells. The work presented here adds to the growing body of research suggesting that polarized cell systems are more suitable for the study of HCV infection and dynamics than non-polarized systems.
DOI: 10.1016/j.jcis.2021.02.067
Journal: Journal of Colloid and Interface Science
PubMed URL: 33677197
Type: Journal Article
Subjects: Atomic force microscopy
Enveloped virus-like particles
Vaccine development
Appears in Collections:Journal articles

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