Project description:Purpose: This goal of this study was to explore the host transcriptomic responses in African swine fever virus experimentally infected pigs using RNA-Sequencing. Methods: RNAs acquired from ten different organ tissue samples were sequenced. Sequencing reads were preprocessed, aligned with the reference genome, assembled and expressions were estimated through bioinformatics approaches. Result: Several uprugulated DEGs were identified. Conclusion: We found important candidate genes and pathways for further testing in African swine fever virus infection in pig.
Project description:African swine fever virus (ASFV) is a large, icosahedral, double-stranded DNA virus in the Asfarviridae family and the causative agent of African swine fever (ASF). ASFV causes a hemorrhagic fever with high mortality rates in domestic and wild pigs. ASFV contains an open reading frame named EP152R, previous research has shown that EP152R is an essential gene for virusrescue in swine macrophages. However, the detailed functions of ASFV EP152R remain elusive. Herein, we demonstrate that EP152R, a membrane protein located in the endoplasmic reticulum (ER), induces ER stress and swelling, triggering the PERK/eIF2α pathway and broadly inhibiting host protein synthesis in vitro. Additionally, EP152R strongly promotes immune evasion, reduces cell proliferation, and alters cellular metabolism. These results suggest that ASFV EP152R plays a critical role in the intracellular environment, facilitating viral replication. Furthermore, virus-level experiments have shown that the knockdown of EP152R or PERK inhibitors efficiently affects viral replication by decreasing viral gene expression. In summary, these findings reveal a series of novel functions of ASFV EP152R and have important implications for understanding host-pathogen interactions.
Project description:Quantitiative analysis of proteomes of porcine macrophages and the stable cell line WSL infected with African swine fever virus using a nanoLC MALDI-Tof/Tof MS platform.
The impact of the infection on the two cell types was analyzed using Gene Ontology term and KEGG pathway enrichment analysis.
Project description:African swine fever is a viral disease of swine caused by the African swine fever virus (ASFV). Currently, ASFV is a serious threat to the global pig industry. A viral strategy to undermine host cell response is to establish a global shutoff of host protein synthesis (virus-induced shutoff, vhs). Here, we characterize ASFV-induced shutoff in primary porcine macrophages by measurement of relative protein synthesis rates based on stable isotope labeling with amino acids in cell culture (SILAC). The impact of ASFV infection on the synthesis of >2000 individual host proteins showed a high degree of variability ranging from complete shutoff to a strong induction of proteins that are absent from naïve cells. By GO-term enrichment analysis the cellular pathways that were most efficiently impacted by vhs were identified. The experimental setup is suitable to quantify vhs after infections with different viruses.
Project description:Long noncoding RNAs (lncRNAs) participate in regulating many biological processes. However, their roles in African swine fever virus(ASFV) pathogenicity are largely unknown. Here, we analyzed the expression profile of lncRNAs and mRNAs in the ASFV-infected or uninfected PAMs by high-throughput sequencing
Project description:In this project we would like to study immune responses to African swine fever virus in pigs. We have realized a large animal experiment using two different viruses and pigs with different immune status. We have collected paxgene blood RNA tubes in order to investigate transcriptional changes at different stages of the early immune response.
