Multiple microRNAs targeted to internal ribosome entry site against foot-and-mouth disease virus infection in vitro and in vivo

Abstract

BackgroundFoot-and-mouth disease virus (FMDV) causes a severe vesicular disease in domestic and wild cloven-hoofed animals. Because of the limited early protection induced by current vaccines, emergency antiviral strategies to control the rapid spread of FMD outbreaks are needed.Here we constructed multiple microRNAs (miRNAs) targeting the internal ribosome entry site (IRES) element of FMDV and investigated the effect of IRES-specific miRNAs on FMDV replication in baby hamster kidney (BHK-21) cells and suckling mice.ResultsFour IRES-specific miRNAs significantly reduced enhanced green fluorescent protein (EGFP) expression from IRES-EGFP reporter plasmids, which were used with each miRNA expression plasmid in co-transfection of BHK-21 cells. Furthermore, treatment of BHK-21 cells with Bi-miRNA (a mixture of two miRNA expression plasmids) and Dual-miRNA (a co-cistronic expression plasmid containing two miRNA hairpin structures) induced more efficient and greater inhibition of EGFP expression than did plasmids carrying single miRNA sequences.Stably transformed BHK-21 cells and goat fibroblasts with an integrating IRES-specific Dual-miRNA were generated, and real-time quantitative RT-PCR showed that the Dual-miRNA was able to effectively inhibit the replication of FMDV (except for the Mya98 strain) in the stably transformed BHK-21 cells.The Dual-miRNA plasmid significantly delayed the deaths of suckling mice challenged with 50× and 100× the 50% lethal dose (LD50) of FMDV vaccine strains of three serotypes (O, A and Asia 1), and induced partial/complete protection against the prevalent PanAsia-1 and Mya98 strains of FMDV serotype O.ConclusionThese data demonstrate that IRES-specific miRNAs can significantly inhibit FMDV infection in vitro and in vivo.

Virus-Specific CD8+ T Cells Infiltrate Melanoma Lesions and Retain Function Independently of PD-1 Expression

“…MCMV-gp100 S27P (MCMV-gp100), MCMV-SL8, WT-MCMV, and MCMV-K181 have all been previously described and were produced and grown as previously described ([22–24]). VacV expressing ovalbumin (VacV-OVA) was kindly provided by Dr. Laurence C. Eisenlohr (The Children’s Hospital of Philadelphia Research Institute and The Perelman School of Medicine at The University of Pennsylvania).…”

Current status and future prospects of yellow fever vaccines

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Global Foot-and-Mouth Disease Research Update and Gap Analysis: 5 - Biotherapeutics and Disinfectants

“…Harnessing the host's own transcriptional silencing mechanisms to prevent FMDV genome replication has been attempted with limited success using short‐interfering RNAs [also called small interfering RNA (siRNA)]; however, micro‐RNAs (miRNAs) have shown some potential. A plasmid encoding dual miRNAs targeting the IRES region proved able to delay or prevent death in suckling mice when co‐administered with high doses of FMDV from multiple serotypes (Chang et al., ). Elsewhere, reduced FMDV attachment to cellular receptors was achieved via the expression of alpha v integrin‐specific miRNAs that inhibited expression of the host cell receptor integrin alpha v, resulting in decreased FMDV infection both in cell culture and in transgenic suckling mice (Du et al., ).…”