Taylor Saunders-Wood (United Kingdom)

University of Cambridge Department of Pathology

Presenter of 1 Presentation

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IN VIVO MODEL FOR PAPILLOMAVIRUS INFECTION AND PREVENTION OF TRANSMISSION AND EFFECTIVE DISINFECTION (ID 1309)

Session Date
07/21/2020
Session Time
10:00 - 17:00
Room
Featured ePosters
Session Type
Poster Viewing - 20-24 July
Session Name
Featured ePosters
Lecture Time
10:06 - 10:07
Presenter
Authors

Abstract

Introduction

Papillomavirus transmission studies have typically used cell-free virus prepared in organotypic raft culture, or recombinant pseudo-virus isolated from monolayer 293TT cells then been investigated by quantitating viral gene transcripts after infecting reporter cells. During natural in vivo infection however, virions are shed from the epithelial surface in squames, and the successful transmission/infection results in new lesion formation.

Methods

We established an in vivo model, which mimics natural papillomavirus transmission/infection, and evaluated them quantitatively by utilising RNAscope to identify lesion formation microscopically and visual inspection for lesions macroscopically, in conjunction with in vitro infection of reporter cells using HPV16/18 raft virus and HPV16 pseudo-virus.

Results

Similar to HPV, MmuPV1 virus titre/infectivity can be quantified by RT-qPCR (E1^E4) or RNAscope (E6E7) using HaCaT cells. Virus titre/infectivity can be also quantified as periods of lesion formation in vivo model with a 6-log dynamic range. Up to 10 million virus particles can be produced from the surface layers of a productive lesion and transmitted with an approximate one log drop in infectivity if transmission is mediated indirectly on fomites. Virus in squames is stable following desiccation, with minimal loss of titre on fomites over 12 months. In contrast, cell-free MmuPV1 virion is not stable with loss of titre within 8 weeks. The efficacy of disinfectants was also evaluated using these in vitro/vivo models.

Conclusions

MmuPV1 in vitro and in vivo infection models using natural viruses produced from productively infected lesions are powerful methods to investigate PV transmission and susceptibility of PV for disinfectants, which can be utilised for HPV transmission. These models have demonstrated different viral shedding patterns that can affect virus stability and susceptibility for disinfectants. Importantly, we have utilised these models to demonstrate the utility of OPA in stark contrast to other studies.

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Moderator of 1 Session

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Basic Science Oral Session

00810 - Oral Session 5: Other Anatomical Sites and Diseases (ID 26)

Session Type
Basic Science Oral Session
Session Date
07/23/2020
Session Time
07:15 - 08:30
Room
Hall D
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