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Scientists design new skin cell culture technique to study human papillomavirus

Novel approach enables genetic analysis of entire viral life cycle, starting with initial infection

PLOS

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IMAGE: This is genome amplification and capsid protein expression in HPV16-infected HFK subjected to organotypic raft culture. view more 

Credit: Bienkowska-Haba M, et al. (2018)

A new cell culture strategy promises to illuminate the mysterious early stages of human papillomavirus (HPV) infection, according to research published in PLOS Pathogens by Malgorzata Bienkowska-Haba and colleagues at Louisiana State University.

Most HPV infections cause no symptoms, but certain types of HPV are associated with cervical, throat, and other cancers. HPV infects skin cells known as keratinocytes before they have fully matured, or differentiated, and are still dividing into new cells. During the final stages of differentiation, uninfected cells cease division, but HPV makes infected cells keep dividing.

For years, details of the early stages of HPV infection have remained murky because scientists have struggled to achieve high enough initial infection rates in keratinocytes in the lab. Thus, only cells in which infection was already well established could be studied. The new approach developed by Bienkowska-Haba and colleagues enables high infection rates of keratinocytes by HPV16, an HPV type that is particularly strongly linked with cancer.

The research team took inspiration from recent reports that, before entering a keratinocyte, HPV16 binds to the extracellular matrix--a mesh of proteins and carbohydrates outside of the cell that performs a variety of structural and molecular functions. They found that high infection rates could be achieved by incubating HPV16 with extracellular matrix materials in a dish, and then seeding it with keratinocytes.

The researchers then demonstrated the potential of this new approach to reveal important genetic information about early HPV infection. They analyzed RNA from infected cells at different time points to determine which HPV genes were expressed and when. They also infected cells with mutant HPV16 viruses lacking different genes, revealing which viral proteins were essential to establish infection.

It is unclear exactly why pre-binding to the extracellular matrix is more effective for establishing infection than introducing HPV16 directly into keratinocytes. Nonetheless, with continued refinement and research, the new approach could help uncover important insights into HPV infection and how it can lead to cancer.

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In your coverage please use this URL to provide access to the freely available article in PLOS Pathogens: http://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1006846

Citation: Bienkowska-Haba M, Luszczek W, Myers JE, Keiffer TR, DiGiuseppe S, Polk P, et al. (2018) A new cell culture model to genetically dissect the complete human papillomavirus life cycle. PLoS Pathog 14(3): e1006846. https://doi.org/10.1371/journal.ppat.1006846

Funding: Funding was provided by the National Institute of Health through grants R01AI081809; R01AI118904; R01DE025565, R01CA211576, and P30GM110703). The Feist Weiller Cancer Center, Louisiana State University Health Sciences Center-Shreveport provided funds for a predoctoral fellowship awarded to SD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing Interests: The authors have declared that no competing interests exist.

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