Cell Lines for the Development of African Swine Fever Virus Vaccine Candidates: An Update

Vaccines, Год журнала: 2022, Номер 10(5), С. 707 - 707

Опубликована: Апрель 29, 2022

African swine fever virus (ASFV) is the etiological agent of a highly lethal disease in both domestic and wild pigs. The has rapidly spread worldwide no available licensed vaccine. An obstacle to construction safe efficient vaccine lack suitable cell line for ASFV isolation propagation. Macrophages are main targets ASFV, they have been widely used study virus–host interactions; nevertheless, obtaining these cells time-consuming expensive, not ethically production large-scale vaccines. To overcome issues, different virulent field isolates adapted on monkey or human continuous lines; however, several culture passages often lead significant genetic modifications loss immunogenicity strain. Thus, groups attempted establish porcine able sustain growth. Preliminary data suggested that some lines might be an alternative primary macrophages research production, although further studies still needed. In this review, we summarize investigate most

Язык: Английский

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Development of an indirect ELISA against African swine fever virus using two recombinant antigens, partial p22 and p30

Journal of Virological Methods, Год журнала: 2022, Номер 309, С. 114611 - 114611

Опубликована: Сен. 2, 2022

African swine fever (ASF) is a highly fatal viral disease affecting pigs. It caused by the ASF virus (ASFV), and causes serious economic losses to industry worldwide, including in Korea. Commercially available enzyme-linked immunosorbent assay (ELISA) kits for detecting anti-ASFV antibodies are used diagnosis surveillance of ASF. In this study, an ELISA was developed detect using two recombinant proteins, p22 p30, from genotype II ASFV. Recombinant transmembrane domain-deleted (p22∆TM) p30 were expressed E.coli vector system pET32a mixed use as antigens indirect ELISA. The p22∆TM/p30-based validated 31 sera I ASFV-infected pigs 1133 uninfected Area under curve test 0.999 [95 % concentration interval 0.992–1.000], sensitivity specificity 93.5 99.8 %, respectively. between run coefficient variation internal quality control serum 6.61 %. seroconversion analysis, showed equal or better ability experimentally challenged with ASFV p72 genotypes (p < 0.05). conclusion, reliable diagnostic method antibodies.

Язык: Английский

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FoxJ1 inhibits African swine fever virus replication and viral S273R protein decreases the expression of FoxJ1 to impair its antiviral effect

Virologica Sinica, Год журнала: 2022, Номер 37(3), С. 445 - 454

Опубликована: Май 2, 2022

African swine fever (ASF) is a highly pathogenic infectious disease that affects domestic pigs and wild boar, which caused by the virus (ASFV). ASF has huge economic losses to pig industry seriously threatens global food security livestock health. To date, there no safe effective commercial vaccine against ASF. Unveiling underlying mechanisms of ASFV-host interplay critical for developing vaccines drugs ASFV. In present study, RNA-sequencing, RT-qPCR Western blotting analysis revealed transcriptional protein levels host factor FoxJ1 were significantly down-regulated in primary porcine alveolar macrophages (PAMs) infected showed overexpression upregulated transcription type I interferon stimulating genes (ISGs) induced poly(dA:dT). function positively regulate innate immune response, therefore, suppressing replication addition, indicated degraded ASFV MGF505-2R E165R proteins through autophagy pathway. Meanwhile, S273R inhibited expression FoxJ1. Altogether, we determined plays an antiviral role replication, impairs FoxJ1-mediated effect degradation Our findings provide new insights into FoxJ1, might help design or infection.

Язык: Английский

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African Swine Fever Virus MGF110-7L Induces Host Cell Translation Suppression and Stress Granule Formation by Activating the PERK/PKR-eIF2α Pathway

Microbiology Spectrum, Год журнала: 2022, Номер 10(6)

Опубликована: Ноя. 15, 2022

African swine fever (ASF) is a highly contagious and often lethal disease of pigs caused by ASF virus (ASFV) recognized as the biggest killer in global industry. Despite exhibiting incredible self-sufficiency, ASFV remains unconditionally dependent on host translation machinery for its mRNA translation. However, less yet known regarding how ASFV-encoded proteins regulate infected cells. Here, we examined interacts with eukaryotic initiation factor 2α (eIF2α) signaling axis, which directs control adaptation to cellular stress. We found that MGF110-7L, previously uncharacterized member multigene family 110, remarkably enhanced phosphorylation level eIF2α. In porcine alveolar macrophage 3D4/21 kidney-15 cells, MGF110-7L triggered eIF2α integrated stress response, resulting suppression formation granules (SGs). Mechanistically, MGF110-7L-induced was mediated via protein kinase R (PKR) PKR-like endoplasmic reticulum (ER) (PERK), this process essential repression SG formation. Notably, our subsequent analyses confirmed overwhelmingly retained ER specific reorganization secretory pathway. Further proteomic biochemical experiments revealed could trigger activate unfolded thus contributing reprogramming. Overall, study both identifies novel mechanism subverts synthesis provides further insights into regulation occurs during infection. IMPORTANCE has become socioeconomic burden threat food security biodiversity, but no commercial vaccines or antivirals are available currently. Understanding viral strategies subvert infection potentially lead new antiviral therapies. study, dissected axis controlling translational reprogramming, addressed role induction responses, phosphorylation, suppression, granule These results define several molecular interfaces cell translation, may guide research dissection pathogenesis.

Язык: Английский

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Deletions of MGF110-9L and MGF360-9L from African swine fever virus are highly attenuated in swine and confer protection against homologous challenge

Journal of Biological Chemistry, Год журнала: 2023, Номер 299(6), С. 104767 - 104767

Опубликована: Май 2, 2023

African swine fever, caused by a large icosahedral DNA virus (African fever virus, ASFV), is highly contagious disease in domestic and feral swine, thus posing significant economic threat to the global industry. Currently, there are no effective vaccines or available methods control ASFV infection. Attenuated live viruses with deleted virulence factors considered be most promising vaccine candidates; however, mechanism which these attenuated confer protection unclear. Here, we used Chinese CN/GS/2018 as backbone homologous recombination generate MGF110-9L MGF360-9L, two genes antagonize host innate antiviral immune response, were (ASFV-ΔMGF110/360-9L). This genetically modified was pigs provided of against parental challenge. Importantly, found ASFV-ΔMGF110/360-9L infection induced higher expression Toll-like receptor 2 (TLR2) mRNA compared determined RNA-Seq RT-PCR analysis. Further immunoblotting results showed that inhibited Pam3CSK4-triggered activating phosphorylation proinflammatory transcription factor NF-κB subunit p65 inhibitor IκBα levels, although activation ASFV-ΔMGF110/360-9L-infected cells ASFV-infected cells. Additionally, show overexpression TLR2 replication p72 protein, whereas knockdown had opposite effect. Our findings suggest might mediated increased signaling.

Язык: Английский

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DDX20: A Multifunctional Complex Protein

Molecules, Год журнала: 2023, Номер 28(20), С. 7198 - 7198

Опубликована: Окт. 20, 2023

DEAD-box decapping enzyme 20 (DDX20) is a putative RNA-decapping that can be identified by the conserved motif Asp-Glu-Ala-Asp (DEAD). Cellular processes involve numerous RNA secondary structure alterations, including translation initiation, nuclear and mitochondrial splicing, assembly of ribosomes spliceosomes. DDX20 reportedly plays an important role in cellular transcription post-transcriptional modifications. On one hand, interact with various factors repress transcriptional process. other forms survival motor neuron complex participates snRNP, ultimately affecting splicing Finally, potentially rely on its RNA-unwinding function to participate microRNA (miRNA) maturation act as component RNA-induced silencing complex. In addition, although not key innate immune system signaling pathway, it affect factor kappa B (NF-κB) p53 pathways. particular, different roles tumorigenesis development through NF-κB pathway. This process regulated such miRNA. influence viral replication cells interacting two proteins Epstein-Barr virus regulate several viruses system, indicating system. Herein, we review effects modification processes, based which provide outlook future research immunity infections.

Язык: Английский

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A porcine kidney-derived clonal cell line with clear genetic annotation is highly susceptible to African swine fever virus

Veterinary Research, Год журнала: 2024, Номер 55(1)

Опубликована: Апрель 4, 2024

Abstract African Swine Fever virus (ASFV), the causative agent of swine fever, is a highly lethal hemorrhagic affecting domestic pigs and wild boars. The primary target cells for ASFV infection are porcine alveolar macrophages (PAMs), which difficult to obtain maintain in vitro, less subjective genetic editing. To overcome these issues facilitate research, we obtained subclonal cell line PK1-C5 by subcloning LLC-PK1 that support stable proliferation. This consequential exhibited high levels similar viral growth characteristics PAMs, while also allowing high-efficiency genomic editing through transfection or lentivirus transduction Cas9. Taken together, our study provided valuable tool research aspects including ASFV-host interactions, pathogenicity, vaccine development.

Язык: Английский

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Repurposing major metabolites of lamiaceae family as potential inhibitors of α-synuclein aggregation to alleviate neurodegenerative diseases: an in silico approach

Frontiers in Pharmacology, Год журнала: 2025, Номер 16

Опубликована: Апрель 16, 2025

Neurodegenerative disorders (NDs) are typically characterized by progressive loss of neuronal function and the deposition misfolded proteins in brain peripheral organs. They molecularly classified based on specific involved, underscoring critical role protein-processing systems their pathogenesis. Alpha-synuclein (α-syn) is a neural protein that crucial initiating progressing various NDs directly or indirectly regulating other ND-associated proteins. Therefore, reducing α-syn aggregation can be an excellent option for combating ND initiation progression. This study presents silico phytochemical-based approach discovering novel neuroprotective agents from bioactive compounds Lamiaceae family, highlighting potential computational methods such as functional networking, pathway enrichment analysis, molecular docking, simulation therapeutic discovery. Functional network analysis established direct indirect involvement NDs. Furthermore, docking interaction studies were conducted to screen 85 major family against aggregation. The results showed five (α-copaene, γ-eudesmol, carnosol, cedryl acetate, spathulenol) had high binding affinity towards with inhibitory activity its MD simulations validated stability interactions determined docking. In addition, pharmacokinetic underscores promising drug candidates, demonstrating blood-brain barrier (BBB) permeability, bioactivity, reduced toxicity. summary, this identifies most suitable targeting recommends these NDs, pending further vitro vivo validation.

Язык: Английский

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PI3K–Akt pathway-independent PIK3AP1 identified as a replication inhibitor of the African swine fever virus based on iTRAQ proteomic analysis

Virus Research, Год журнала: 2023, Номер 327, С. 199052 - 199052

Опубликована: Фев. 15, 2023

African swine fever (ASF) is a severe infectious disease that has high global prevalence. The fatality rate of pigs infected with ASF virus (ASFV) close to 100%; in the absence safe and reliable commercial vaccine, this poses threat pig industry public health. To better understand interaction ASFV its host, isobaric tags for relative absolute quantitation combined liquid chromatography-mass spectrometry was used conduct quantitative proteomic analysis bone marrow-derived macrophage cells ASFV. Overall, 4579 proteins were identified; 286 these significantly upregulated 69 downregulated after infection. Gene Ontology, Kyoto Encyclopedia Genes Genomes, protein-protein network analyses obtain insights into dynamics complexity ASFV-host interaction. In addition, immunoblotting revealed expression PIK3AP1, RNF114, FABP5 TRAM1 Overexpression PIK3AP1 RNF114 inhibited replication vitro, but suppressive effect on independent PI3K-Akt pathway. Further studies confirmed MGF360-9L interacts reduce protein level. Moreover, LY294002, an inhibitor pathway, replication, indicating importance pathway This study identified interactions between host provides reference development anti-ASFV strategies studying potential mechanisms pathogenesis

Язык: Английский

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Identification and analysis of the interaction network of African swine fever virus D1133L with host proteins

Frontiers in Microbiology, Год журнала: 2022, Номер 13

Опубликована: Ноя. 3, 2022

African swine fever (ASF) is a contagious and lethal hemorrhagic disease in pigs; its spread results huge economic losses to the global pig industry. ASF virus (ASFV) large double-stranded DNA encoding >150 open reading frames. Among them, ASFV-encoded D1133L was predicted be helicase but specific function remains unknown. Since virus-host protein interactions are key understanding viral function, we used co-immunoprecipitation combined with liquid chromatography-mass spectrometry investigate D1133L. This study describes interaction network of ASFV porcine kidney PK-15 cells. Overall, 1,471 host proteins that potentially interact identified. Based on these proteins, protein-protein constructed. Gene ontology Kyoto Encyclopedia Genes Genomes enrichment analyses showed cellular D1133L-interacted involved ribosome, spliceosome, RNA transport, oxidative phosphorylation, proteasome, replication. Vimentin (VIM), tripartite motif-containing 21 (TRIM21), Tu translation elongation factor (TUFM) were confirmed vitro. VIM or TRIM21 overexpression significantly promoted replication, TUFM inhibited These help elucidate functions potential mechanisms underlying

Язык: Английский

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