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Research Project: Japanese Encephalitis Virus Prevention and Mitigation Strategies

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Title: Porcine macrophage-like cell line C Delta 2+ is susceptible to Japanese encephalitis virus infection

Author
item ADETUNJI, SHAKIRAT - Kansas State University
item Smolensky, Dmitriy
item Mitzel, Dana
item Chitko-Mckown, Carol
item CERNICCHIARO, NATALIA - Kansas State University
item Noronha, Leela

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 2/5/2021
Publication Date: N/A
Citation: N/A

Interpretive Summary: Interpretive Summary not required in accordance with ARS-115 Publications P & P 152.1 v.5 (10/19/2019)chapter 5 page 31 Matrix for Data Entry Determinations.kmm

Technical Abstract: Introduction: Japanese encephalitis virus (JEV) is a zoonotic arthropod-borne flavivirus that is a leading cause of severe neurologic infection in humans. Pigs have a lasting viremia upon natural infection making them important reservoirs of JEV. To understand JEV pathogenesis in pigs, we examined the susceptibility of an established porcine monocyte-derived macrophage-like cell line (C Delta 2+) to the attenuated JEV strain, SA-14-14-2. Methods: Monolayers of C Delta 2+ and BHK-21 (positive control) cells were infected with SA-14-14-2 for 5 days at a multiplicity of infection (MOI) of 0.1. Culture supernatants and cells were collected from 0 - 120 hours post infection (hpi), and monolayers were observed for cytopathic effects (CPE). Infectious virus in supernatants was quantified using plaque assays and cells were stained with trypan blue to determine viability. An indirect immunofluorescence assay was used to detect JEV-NS1 antigens. Results: C Delta 2+ cells were susceptible to SA-14-14-2 and produced infectious virus with a mean peak titer comparable to BHK-21 cells which are known to be vulnerable to JEV. The proportion of viable C Delta 2+ and BHK-21 cells also declined at a similar rate. Infected C Delta 2+ and BHK cells showed time-dependent CPE and intracellular localization of the JEV-NS1 protein was observed at 24hpi. Conclusions: These findings demonstrate that C Delta 2+ represents a relevant cell line for understanding JEV infection dynamics in a natural host species. These data provide a foundation to compare various JEV strains in vitro to allow for better understanding of host cell mechanisms critical for viral replication and maintenance in pigs.