Development and in vivo evaluation of MGF100-1R deletion mutant in an African swine fever virus Chinese strain

Liu, Yingnan; Yao, Li; Xie, Zhixun; Ao, Qingying; Di, Dongdong; Yu, Wanqi; Lv, Lu; Zhong, Qiuping; Song, Yingying; Liao, Xinxin; Song, Qing; Wang, Heng; Chen, Hongjun

doi:10.1016/j.vetmic.2021.109208

Cited by 14 publications

(15 citation statements)

References 22 publications

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“…Interestingly, loss of MGF 100 members was observed during the process of adapting a virulent Georgia strain to grow in cultured cell lines ( 60 ). Deletion of MGF 100-1R, from a virulent genotype II Chinese strain ( 73 ), or of l8L from Georgia 2010 was shown not to reduce virulence of the virus in pigs or reduce virus replication in porcine macrophages ( 74 ). However, simultaneous deletion of genes l7L, l8L, l9L, l10L, and l11L from a Chinese virulent isolate reduced virulence, and surviving pigs were protected against challenge ( 59 ).…”

Section: Discussionmentioning

confidence: 99%

African Swine Fever Virus and Host Response: Transcriptome Profiling of the Georgia 2007/1 Strain and Porcine Macrophages

Cackett

Portugal

Matelska

et al. 2022

J Virol

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African swine fever virus (ASFV) has a major global economic impact. With a case fatality in domestic pigs approaching 100%, it currently presents the largest threat to animal farming. Although genomic differences between attenuated and highly virulent ASFV strains have been identified, the molecular determinants for virulence at the level of gene expression have remained opaque. Here we characterise the transcriptome of ASFV genotype II Georgia 2007/1 (GRG) during infection of the physiologically relevant host cells, porcine macrophages. In this study we applied Cap Analysis Gene Expression sequencing (CAGE-seq) to map the 5’ ends of viral mRNAs at 5 and 16 hours post-infection. A bioinformatics analysis of the sequence context surrounding the transcription start sites (TSSs) enabled us to characterise the global early and late promoter landscape of GRG. We compared transcriptome maps of the GRG isolate and the lab-attenuated BA71V strain that highlighted GRG virulent-specific transcripts belonging to multigene families, including two predicted MGF 100 genes I7L and I8L. In parallel, we monitored transcriptome changes in the infected host macrophage cells. Of the 9,384 macrophage genes studied, transcripts for 652 host genes were differentially regulated between 5 and 16 hours-post-infection compared with only 25 between uninfected cells and 5 hours post-infection. NF-kB activated genes and lysosome components like S100 were upregulated, and chemokines such as CCL24, CXCL2, CXCL5 and CXCL8 downregulated. African swine fever virus (ASFV) causes haemorrhagic fever in domestic pigs with case fatality rates approaching 100%, and no approved vaccines or antivirals. The highly-virulent ASFV Georgia 2007/1 strain (GRG) was the first isolated when ASFV spread from Africa to the Caucasus region in 2007. Then spreading through Eastern Europe, and more recently across Asia. We used an RNA-based next generation sequencing technique called CAGE-seq to map the starts of viral genes across the GRG DNA genome. This has allowed us to investigate which viral genes are expressed during early or late stages of infection and how this is controlled, comparing their expression to the non-virulent ASFV-BA71V strain to identify key genes that play a role in virulence. In parallel we investigated how host cells respond to infection, which revealed how the ASFV suppresses components of the host immune response to ultimately win the arms race against its porcine host.

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Section: Discussionmentioning

confidence: 99%

African Swine Fever Virus and Host Response: Transcriptome Profiling of the Georgia 2007/1 Strain and Porcine Macrophages

Cackett

Portugal

Matelska

et al. 2022

J Virol

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“…Primary bone marrow-derived macrophages (BMDMs) were collected from specific-pathogen-free (SPF) pigs as described previously (Liu et al, 2021). BMDMs cells were cultured in RPMI 1640 (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 10 % fetal bovine serum (FBS), 2 mM L-glutamine (Gibco, Thermo Fisher Scientific, Waltham, MA, USA), 1% Penstrep and 10 ng/ml rpGM-CSF (R&D Systems, Minneapolis, MN, USA).…”

Section: Virus and Cellsmentioning

confidence: 99%

“…The genes were knocked out and generated by homologous recombination by infection with parental virus GZ201801 and transfection with the recombination transfer vectors (p72eGFP EP153R/ EP402R and p72mCherry MGF_360-12L/13L/14L) described in our previous study (Liu et al, 2021). The recombinant transfer vector p72eGFP EP153R/ EP402R contains the upstream and downstream fragments (about 1,200 bp each) of the target deletion EP153R-EP402R gene with the p72 promoter and eGFP gene in the middle (Borca et al, 2017).…”

Section: Construction Of Recombinant Virusmentioning

confidence: 99%

“…Two groups of pigs (n = 5 per group) were infected with the ASFV-ECM3 virus or the WT virus intramuscularly (IM), both at a dose of 10 5 TCID 50 . ASFV-inoculated pigs were monitored for rectal temperature as described previously (Liu et al, 2021). Clinical samples of sera, heparin-anticoagulated whole blood, oral swabs, nasal swabs, and anal swabs were collected at 0, 3, 7, 10, 14, 21, and 28 dpi.…”

Section: Animal Experimentsmentioning

confidence: 99%

“…The organs and tissues of ASFV-ECM3 virus-infected pigs euthanized were collected, and the viral distribution in pigs was analyzed. Organs and tissues from the dead pigs and surviving pigs that were euthanized at the end of the observation period were scored using the gross scoring system, and 100 mg of the indicated samples were homogenized by Tissuelyser-FEII (Shanghai Jingxin Co. Ltd., Shanghai, China) in 800 µl PBS supplemented with 1% Pen-strep for the detection of virus titer by TCID 50 assay as described previously (Liu et al, 2021).…”

Section: Animal Experimentsmentioning

confidence: 99%

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Protection Evaluation of a Five-Gene-Deleted African Swine Fever Virus Vaccine Candidate Against Homologous Challenge

Xie

Liu

Di³

et al. 2022

Front. Microbiol.

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African swine fever virus (ASFV) represents a serious threat to the global swine industry, and there are no safe or commercially available vaccines. Previous studies have demonstrated that inactivated vaccines do not provide sufficient protection against ASFV and that attenuated vaccines are effective, but raise safety concerns. Here, we first constructed a deletion mutant in which EP153R and EP402R gene clusters were knocked out. Based on the deletion mutant, a further deletion from the MGF_360-12L, MGF_360-13L to MGF_360-14L genes was obtained. The five-genes knockout virus was designated as ASFV-ΔECM3. To investigate the efficacy and safety of the ASFV-ΔECM3 virus as a vaccine candidate, the evaluation of the virus was subsequently carried out in pigs. The results showed that the ASFV-ΔECM3 virus could induce homologous protection against the parental isolate, and no significant clinical signs or viremia were observed. These results show that the contiguous deletion mutant, ASFV-ΔECM3 encompassing the EP153R/EP402R and MGF_360-12L/13L/14L genes, could be a potential live-attenuated vaccine candidate for the prevention of ASFV infection.

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Identification of monoclonal antibody targeting epitope on p72 trimeric spike of African swine fever virus

et al. 2023

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Development and in vivo evaluation of MGF100-1R deletion mutant in an African swine fever virus Chinese strain

Cited by 14 publications

References 22 publications

African Swine Fever Virus and Host Response: Transcriptome Profiling of the Georgia 2007/1 Strain and Porcine Macrophages

African Swine Fever Virus and Host Response: Transcriptome Profiling of the Georgia 2007/1 Strain and Porcine Macrophages

Protection Evaluation of a Five-Gene-Deleted African Swine Fever Virus Vaccine Candidate Against Homologous Challenge

Identification of monoclonal antibody targeting epitope on p72 trimeric spike of African swine fever virus

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