Multiplex Profiling of Cytokines released by A549 Human Lung Epithelial Cells after Infection with Encephalomyocarditis Virus and Vesicular Stomatitis Virus


The model cell-virus system of the human lung epithelial cell line A549:Encephalomyocarditis Virus (EMCV) and A549:Vesicular Stomatitis Virus (VSV) have been used routinely in standard cytopathic effect inhibition (CPE) assays for measuring the bioactivity of interferons (IFNs) and neutralizing titers of anti-IFN antibodies. In this study, we have used solid phase single-and multiplex ELISAs to examine a panel of cytokines including IFN-α, IFN-β, IFN-γ. IFN-λ1/IL-29, IFN-λ2/IL-28A, IFN-ω, selected interleukins (IL), and TNF in media collected at 4, 8, 12, 24, 48, and 72 hours after exposure to EMCV or VSV. Infection of A549 cells with EMCV over a near five long range of MOI elicited substantial increases (vs. non-infected cells) in IFN-α and IFN-β in A549 media. EMCV also increased induction of IL-6 and markedly elevated both IFN-λ1 and IFN-λ2. In contrast, infection with similar MOI of VSV failed to elicit increased induction of IL-6, IFN-α, IFN-β, IFN-λ1, and IFN-λ2. Neither virus produced detectable levels of IFN-γ, TNF-α, TNF-β, IL-1α, IL-1β, IL-12, IL-13, IL-17, or IL-23 at any time point examined. Notably, A549 cells were more sensitive to VSV than to EMCV at similar MOIs at time points over 24 hrs post inoculation. In conclusion, this study provides unique insight into the cytokine (protein) production profiles of A549 cells treated with VSV and EMCV allowing a view of both the similarities and major differences in cytokine induction generated by these hallmark negative- and positive-strand RNA viruses (respectively) in culture.


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Multiplex Profiling of Cytokines released by A549

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