Coxsackievirus B3 Infection of Human Neural Progenitor Cells Results in Distinct Expression Patterns of Innate Immune Genes

Abstract

Coxsackievirus B3 (CVB3), a member of Picornaviridae family, is an important human pathogen that causes a wide range of diseases, including myocarditis, pancreatitis, and meningitis. Although CVB3 has been well demonstrated to target murine neural progenitor cells (NPCs), gene expression profiles of CVB3-infected human NPCs (hNPCs) has not been fully explored. To characterize the molecular signatures and complexity of CVB3-mediated host cellular responses in hNPCs, we performed QuantSeq 3′ mRNA sequencing. Increased expression levels of viral RNA sensors (RIG-I, MDA5) and interferon-stimulated genes, such as IFN-β, IP-10, ISG15, OAS1, OAS2, Mx2, were detected in response to CVB3 infection, while IFN-γ expression level was significantly downregulated in hNPCs. Consistent with the gene expression profile, CVB3 infection led to enhanced secretion of inflammatory cytokines and chemokines, such as interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemoattractant protein-1 (MCP-1). Furthermore, we show that type I interferon (IFN) treatment in hNPCs leads to significant attenuation of CVB3 RNA copy numbers, whereas, type II IFN (IFN-γ) treatment enhances CVB3 replication and upregulates suppressor of cytokine signaling 1/3 (SOCS) expression levels. Taken together, our results demonstrate the distinct molecular patterns of cellular responses to CVB3 infection in hNPCs and the pro-viral function of IFN-γ via the modulation of SOCS expression.

Immune Transcriptome of Cells Infected with Enterovirus Strains Obtained from Cases of Type 1 Diabetes

“…Previous studies of acute enteroviral infection showed that: (a) cultured neural cells infected with CBV3 had enhanced secretion of IL6, IL8, and MCP1 and the upregulation of SOCS3, a suppressor of cytokine signaling [ 26 ]; (b) human pancreatic islets infected with CVB3 and CVB4 secreted high amounts of IL6, IL8, MIP-alpha and -beta, and TNF; (c) human pancreatic beta cells infected with CBV4 showed the activation of innate immune pathways including the expression of IFN (alpha, beta, lambda), IL6, IL8, and IL27 and also showed a defective insulin response to glucose [ 35 ]; (d) patients with hand-foot-and-mouth disease caused by EV-A71 and EV-A6 had significantly increased serum levels of IL5, IL7, IL13, IL16, IL18, IP10, NGF-beta, PDGF-beta, and TGFB1 [ 36 ].…”

“…CBV3 was used as a reference strain for transcriptome analysis since this virus is pancreotropic and diabetogenic in mice [ 20 , 21 , 22 ] and supported by serologic studies in humans [ 23 ] as well as by the investigation of some T1D cases [ 24 ]. Regarding transcriptome analysis, CBV3 was chosen since complete data have been published on the cellular and transcriptome consequences of CBV3 infection in human cells [ 25 , 26 ]. For acute infection, confluent cultures in T25 flasks were infected at a multiplicity of infection of 3 using 1 mL of virus diluted in cell culture medium and incubated on a rocking platform at 37 °C.…”

A Comparison of Low Read Depth QuantSeq 3′ Sequencing to Total RNA-Seq in FUS Mutant Mice

“…The goal of this article is not to disprove QuantSeq's capability of producing useful results. A demonstration of its utility can be found in Oh et al (2020) which found that the levels of expression of various cytokines correlated with changes in gene expression found using QuantSeq. As with all methods though, we advise care is taken and validation is performed before any conclusions are reached.…”

mRNA and miRNA profiling of Zika virus-infected human umbilical cord mesenchymal stem cells identifies miR-142-5p as an antiviral factor

“…Total RNA was isolated using Direct-zol RNA mini Prep Kit (Zymo Research, Germany) following manufacturer's instructions. RNA sequencing was performed using an Illumina HiSeq 2500 sequencer (Illumina, San Diego, CA) as previously described [18,23,24]. Sequence reads were aligned to the reference genome (Gene bank accession no.…”