Cells infected with human papilloma pseudovirus display nuclear reorganization and heterogenous infection kinetics

Date
2022-06-06
Advisor
Instructor
Source Title
Cytometry
Print ISSN
1552-4922
Electronic ISSN
1552-4930
Publisher
John Wiley & Sons, Ltd
Volume
101
Issue
12
Pages
1035 - 1048
Language
English
Type
Article
Journal Title
Journal ISSN
Volume Title
Abstract

Human papillomaviruses (HPV) are small, non-enveloped DNA viruses, which upon chronic infection can provoke cervical and head-and-neck cancers. Although the infectious life cycle of HPV has been studied and a vaccine is available for the most prevalent cancer-causing HPV types, there are no antiviral agents to treat infected patients. Hence, there is a need for novel therapeutic entry points and a means to identify them. In this work, we have used high-content microscopy to quantitatively investigate the early phase of HPV infection. Human cervical cancer cells and immortalized keratinocytes were exposed to pseudoviruses (PsV) of the widespread HPV type 16, in which the viral genome was replaced by a pseudogenome encoding a fluorescent reporter protein. Using the fluorescent signal as readout, we measured differences in infection between cell lines, which directly correlated with host cell proliferation rate. Parallel multiparametric analysis of nuclear organization revealed that HPV PsV infection alters nuclear organization and inflates promyelocytic leukemia protein body content, positioning these events at the early stage of HPV infection, upstream of viral replication. Time-resolved analysis revealed a marked heterogeneity in infection kinetics even between two daughter cells, which we attribute to differences in viral load. Consistent with the requirement for mitotic nuclear envelope breakdown, pharmacological inhibition of the cell cycle dramatically blunted infection efficiency. Thus, by systematic image-based single cell analysis, we revealed phenotypic alterations that accompany HPV PsV infection in individual cells, and which may be relevant for therapeutic drug screens.

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Keywords
Cell cycle, High-throughput microscopy, Human papillomavirus, Infection kinetics, Live cell imaging, Nucleus, Pseudovirus
Citation
Published Version (Please cite this version)