Article ; Online: In Vitro Infection Dynamics of Japanese Encephalitis Virus in Established Porcine Cell Lines
Pathogens, Vol 10, Iss 1468, p
2021 Volume 1468
Abstract: Japanese encephalitis virus (JEV) is a zoonotic mosquito-borne pathogen that regularly causes severe neurological disease in humans in Southeast Asia and the Western Pacific region. Pigs are one of the main amplifying hosts of JEV and play a central role ...
| Abstract | Japanese encephalitis virus (JEV) is a zoonotic mosquito-borne pathogen that regularly causes severe neurological disease in humans in Southeast Asia and the Western Pacific region. Pigs are one of the main amplifying hosts of JEV and play a central role in the virus transmission cycle. The objective of this study was to identify in vitro cell systems to investigate early effects of JEV infection including viral replication and host cell death. Here, we demonstrate the susceptibility of several porcine cell lines to the attenuated genotype III JEV strain SA14-14-2. Monolayers of porcine nasal turbinate (PT-K75), kidney (SK-RST), testis (ST), and monocyte-derived macrophage (C <math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mo>Δ</mo></semantics></math> 2+) cells were infected with SA14-14-2 for up to five days at a multiplicity of infection (MOI) of 0.1. The hamster kidney cell line BHK-21, previously shown to be susceptible to SA14-14-2, was used as a positive control. Culture supernatants and cells were collected between 0 and 120 h post infection (hpi), and monolayers were observed for cytopathic effect (CPE) using brightfield microscopy. The number of infectious virus particles was quantified by plaque assay and cell viability was determined using trypan blue staining. An indirect immunofluorescence assay was used to detect the presence of JEV NS1 antigens in cells infected at 1 MOI. All four porcine cell lines demonstrated susceptibility to SA14-14-2 and produced infectious virus by 12 hpi. Virus titers peaked at 48 hpi in C <math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mo>Δ</mo></semantics></math> 2+, BHK-21, and SK-RST cells, at 72 hpi in PT-K75, and at 120 hpi in ST cells. CPE was visible in infected C <math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mo>Δ</mo></semantics></math> 2+ and BHK-21 cells, but not ... |
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| Keywords | arboviruses ; cell culture ; Japanese encephalitis ; infection ; in vitro ; porcine ; Medicine ; R |
| Subject code | 570 |
| Language | English |
| Publishing date | 2021-11-01T00:00:00Z |
| Publisher | MDPI AG |
| Document type | Article ; Online |
| Database | BASE - Bielefeld Academic Search Engine (life sciences selection) |
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