ABSTRACT: Development of a new internally controlled one-step real-time RT-PCR for the molecular detection of enterovirus A71 in Africa and Madagascar.
Project description:Objective Enterovirus 71 (EV-A71) is a major pathogen of severe hand, foot and mouth disease (HFMD) in children, but the mechanism by which it develops into severe HFMD remains unclear, especially the role of macrophage-mediated immune dysregulation. Methods Macrophages infected with EV-A71 were collected for transcriptomic analysis, and were indirectly co-cultured with nerve cells to observe their inhibitory effects on nerve cells. Results High concentration of EV-A71 infected macrophage supernatant inhibited SH-SY5Y cell proliferation. ENSG00000285779, TICAM2, RPL13AP26 and HNF4G are significantly different in EV-A71 or inactivated EV-A71 infected macrophages than in control. ENSG00000264324, ENSG00000260643, ISLR2, CCR7, TENM4, INO80B-WBP1, BLOC1S5-TXNDC5 are potential genes about direct virus damage or viral RNA recognition in macrophages. GO annotation and KEGG analysis indicate that EV-A71 infection cause the changes of neural receptor-ligand binding pathway, activation of specific immunity, calcium signaling pathway, and cell aggregation. Conclusions Macrophages are activated early during EV-A71 infection, thus initiating specific immunity, which is closely related to the severe HFMD. The nerve damage pathway and calcium signaling pathway caused by EV-A71 virus infection of macrophages deserve to more attention.
Project description:A siRNA screening using a commercially available membrane trafficking library identified Rab11 as a pro-viral host factor in motor neuron NSC34 cell line infected with EV-A71, a major causative agent of Hand, Foot and Mouth Disease (HFMD). This project aims at deciphering the role of Rab11 during EV-A71 infection by using various molecular virology and cell biology approaches. We also identified Rab11 interacting partners during EV-A71 infection by pulldown and mass spectrometry.
Project description:In this study, we aimed to adopt the transcriptome sequencing technology to obtain the different changes of transcriptome profiles after infecting with EV-A71 in human neuroblastoma cell line SH-SY5Y. And then, through systematic bioinformatics analysis, we hope to find useful clues for the pathogenesis of HFMD.
Project description:Non-structural 2B protein of enterovirus-A71 has reported involving in intracellular Ca2+ manipulation and altering cellular homeostasis such as inducing cell death in human SH-SY5Y cells. The aim of the study is to profile transcriptomic signature of human neuroblastoma SH-SY5Y cells altered by EV-A71 2B protein using RNA-sequencing analysis. We generated mRNA expression profiles of SH-SY5Y cells transfected with EV-A71 2B protein fused with mCherry and FLAG tag protein (2BmCherry) and mCherry as well as parental SH-SY5Y cells. We find that 7 genes including CCL2, RELB, IL32, PLAT, PTGES, PHLDA1, and TNFRSF9 are uniquely overexpressed in 2BmCherry comparing to mCherry. Moreover, there were 333 upregulated and 333 downregulated genes showed significant different expression level in 2BmCherry transcriptome in comparison with SHSY5Y transcriptome but not in mCherry vs SHSY5Y comparison. Functional analysis showed that EV-A71 2B upregulated genes involved Ca2+-related signaling pathways participating gene expression, immune response, apoptosis, and long-term potentiation (synaptic adaptation) of neuron in the transfected SH-SY5Y cells.
