A Murine Model of Dengue Virus-induced Acute Viral Encephalitis-like Disease

Summary

Here, we present a protocol for creating an immunocompetent ICR (Institute of Cancer Research) murine model of central nervous system infection to display the development of neuropathy. Monitoring acute viral encephalitic disorders by identical disease scores could be performed for showing dengue virus-induced neuropathy in vivo.

Abstract

Dengue virus (DENV), an arthropod-borne virus transmitted by mosquitoes, may cause the severe disease known as dengue hemorrhagic fever, which is characterized by lethal complications due to plasma leakage, ascites, pleural effusion, respiratory distress, severe bleeding, and organ impairment. A few cases of DENV infection present neurological manifestations; however, studies have not explored DENV-induced neuropathogenesis further. In this study, we present a protocol to use an immunocompetent outbred ICR (Institute of Cancer Research) mouse for investigating the induction of central nervous system (CNS) infection with DENV, followed by the progression of acute viral encephalitis-like disease.

Introduction

DENV, an arthropod-borne virus of the Flaviviridae family, contains a positive-sense RNA genome that encodes three viral structural proteins (capsid, premembrane, and envelope) and seven viral nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5). The four serotypes of DENV (DENV1-4), which infect approximately 390 million people annually, cause a global burden even though governments have directed substantial efforts toward mosquito vector and disease control¹. Currently, protective vaccines and therapeutic antiviral drugs are under development and require further long-term validation². In clinical....

Protocol

Experimental protocols of animal study were approved by the Institutional Animal Care and User Committee of the National Defense Medical Center (IACUC number: 16-261), according to guidelines established by the Ministry of Science and Technology, Taiwan.

1. Infection Procedure

1. Prepare nonadapted DENV2 (strain PL046) stocks⁹ (originally obtained from the Centers for Disease Control in Taiwan, ranging from 2.5 x 10⁷ to 1 x 10⁹ PFU/mL).

Discussion

DENV infection has been detected in the CNS of severe dengue patients^3,17, suggesting the possibility of the manifestation of acute viral encephalitis occurred during dengue pathogenesis. Here, we report an in vivo murine model of DENV infection for studying the involvement of CNS dysfunction in severe dengue, particularly with a focus on DENV-induced acute viral encephalitis-like illnesses. As compared with the previous models, particularly for one-route infecti.......

Materials

Name	Company	Catalog Number	Comments
Roswell Park Memorial Institute 1640 medium (RPMI)	Gibco	11875-085	Diluting virus
0.3-mL Insulin Syringe	BD Ultra-Fine­II	328838	Intracerebral and intraperitoneal injection
Microbalance	METTLER TOLEDO's LabX	AL104	Weight mouse
Non-adapted DENV2 (strain PL046)	The Centers for Disease Control of Taiwan	-	Infect mouse
Institute of Cancer Research (ICR) suckling mouse	BioLASCO Taiwan Co., Ltd	-	Our murine model