Abstract
BackgroundChikungunya virus (CHIKV) is a prevalent mosquito-borne pathogen that is emerging in many parts of the globe causing significant human morbidity. Here, we report the isolation and characterization of an infectious CHIKV from banked serum specimens of suspected patients from the 2009 epidemic in Thailand.MethodsStandard plaque assay was used for CHIKV isolation from the banked serum specimens. Isolated CHIKV was identified base on E1 structural gene sequence. Growth kinetic, infectivity, cell viability and cytokine gene expression throughout CHIKV infection in a permissive cell line, 293T cells, was performed using several approaches, including standard plaque assay, immunofluorescence assay, classical MTT assay, and quantitative real-time PCR. Two tailed Student’s t test was used for evaluation statistically significance between the mean values of the groups.ResultsBased on the E1 structural gene sequence and phylogenetic analysis, we identified the virus as the CHIK/SBY8/10 isolate from Indonesia. Assessment of the growth kinetics, cytopathic effects as well as its ability to induce cellular immune responses suggested that the currently isolated CHIK/SBY8/10 virus is relatively more virulent than a known CHIKV vaccine strain, which also induces more dramatic proinflammatory responses.ConclusionsOur studies further add to the infectivity of a less-studied yet infectious CHIKV isolate as well as underscored the importance and utility of 293T cells as an excellent cell culture model for studying viral growth, CHIKV-induced inflammatory cellular responses and cell death. Together, these studies provide novel information on the CHIKV biology, infectivity and virus-cell interaction, which would help develop novel interventions against the infection.
Highlights
Chikungunya virus (CHIKV) is a prevalent mosquito-borne pathogen that is emerging in many parts of the globe causing significant human morbidity
While A. aegypti was long been considered as the only mosquito vector for CHIKV, investigation in the 2005–2006 epidemic on Reunion island that involved 266,000 human cases suggested that a single mutation, an alanine-to-valine substitution in the E1 envelope glycoprotein gene (E1) envelope glycoprotein at the position 226 enabled virus acquisition by a new mosquito vector, A. albopictus, commonly known as the Asian tiger mosquito, which was implicated in that epidemic [7]
Isolation of a chikungunya virus isolate from human serum To contribute to the biology and virulence of infectious CHIKV, we sought to isolate an infectious strain from human patients
Summary
Chikungunya virus (CHIKV) is a prevalent mosquito-borne pathogen that is emerging in many parts of the globe causing significant human morbidity. First discovered in Tanzania in African continent in 1952 and later identified in Thailand in 1958, Chikungunya virus (CHIKV) fever is a re-emerging public health problem with significant human morbidity, accounting for approximately 1.4 million suspected cases worldwide [1]. The control of the virus during infection is achieved through acquired immune responses; the persistence of the virus in joint tissue is associated with its pro-arthritogenic potential, likely inducing the production of proinflammatory factors [12]. Despite these studies, the mechanisms influencing immunopathogenesis of CHIKV infection remains unclear [13, 14]
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