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:The EV-A71 genome has accumulated mutations since the virus was first detected in 1969. To characterize the polymorphism of the EV-A71 3C protease, we performed RNA sequencing using chimeric EV-A71 strains carrying the 3C protease from a clinical strain. The chimeric EV-A71 was generated using a reverse genetic system and inoculated into RD-S cells. Gene expression profiling analysis of RD-S cells infected with the chimeric EV-A71 strains was performed.
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.
