Modelling Lyssavirus Infections in Human Stem Cell-Derived Neural Cultures.

Vinod Sundaramoorthy,John Bingham,Nathan Godde,Carmel M O'Brien,Diane Green,John M Haynes,Ryan J Farr,Megan Dearnley

doi:10.3390/v12040359

Abstract

Rabies is a zoonotic neurological infection caused by lyssavirus that continues to result in devastating loss of human life. Many aspects of rabies pathogenesis in human neurons are not well understood. Lack of appropriate ex-vivo models for studying rabies infection in human neurons has contributed to this knowledge gap. In this study, we utilize advances in stem cell technology to characterize rabies infection in human stem cell-derived neurons. We show key cellular features of rabies infection in our human neural cultures, including upregulation of inflammatory chemokines, lack of neuronal apoptosis, and axonal transmission of viruses in neuronal networks. In addition, we highlight specific differences in cellular pathogenesis between laboratory-adapted and field strain lyssavirus. This study therefore defines the first stem cell-derived ex-vivo model system to study rabies pathogenesis in human neurons. This new model system demonstrates the potential for enabling an increased understanding of molecular mechanisms in human rabies, which could lead to improved control methods.

Highlights

The genus Lyssavirus, belonging to family Rhabdoviridae, consists of a group of negative strandRNA viruses capable of causing rabies-like disease in humans and other mammals [1]
hESC line WA09 [59] (H9)-NPC and HDF51i-509-NPC cultures were fixed following 21 and 24 days of differentiation respectively, for characterization by immunostaining and confocal microscopy. This demonstrated that both embryonic stem cells (ESC)- and induced pluripotent stem cells (iPSC)-derived neural cultures consisted of mature neurons which positively stained for MAP2, TUJI and NEUN, with GFAP-positive astrocytes
We demonstrate that human ESC- and iPSC-derived neural cultures could be used as effective ex-vivo models to study several pathogenic mechanisms associated with human rabies

Summary

Introduction

The genus Lyssavirus, belonging to family Rhabdoviridae, consists of a group of negative strandRNA viruses capable of causing rabies-like disease in humans and other mammals [1]. The genus Lyssavirus, belonging to family Rhabdoviridae, consists of a group of negative strand. Rabies virus belongs to the prototype Rabies lyssavirus species and usually transmitted between terrestrial carnivore species, to a lesser extent bat species and humans [2]. There are 15 other officially recognized species within the Lyssavirus genus, most of which have been isolated from bats [1,3,4]. The death rate from rabies is high in developing countries due to poor access to PEP and many of the victims are children under the age of 15 years [6,7,8]. Victims are usually infected through the bite of infected animals. Once inside the Viruses 2020, 12, 359; doi:10.3390/v12040359 www.mdpi.com/journal/viruses

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