scholarly journals The A179L Gene of African Swine Fever Virus Suppresses Virus-Induced Apoptosis but Enhances Necroptosis

Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2490
Author(s):  
Jun Shi ◽  
Wei Liu ◽  
Miao Zhang ◽  
Jing Sun ◽  
Xiulong Xu
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Rna Viruses ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Epithelial Cell Line ◽  
L929 Cells ◽  
Tnf Α ◽  
Hsv 1

A179L, a non-structural protein of African swine fever virus (ASFV), is capable of suppressing apoptosis by binding the BH3 domain of the pro-apoptotic Bcl-2 family proteins via a conserved ligand binding groove. Our present study aims to determine if A179L affects necroptosis, the second form of programmed cell death induced by DNA and RNA viruses. Here we report that A179L enhanced TNF-α or TSZ (TNF-α, Smac, and Z-Vad)-induced receptor-interacting protein kinase (RIPK1), RIPK3, and mixed lineage kinase domain like peudokinase (MLKL) phosphorylation in L929 cells, a murine fibrosarcoma cell line. Sytox green staining revealed that A179L significantly increased the number of necroptotic cells in TSZ-treated L929 cells. Using human herpes simplex virus 1 (HSV-1) to model DNA virus-induced cell death, we found that A179L blocked the HSV-1-induced cleavage of poly (ADP-ribose) polymerase (PARP), caspase 8, and caspase 3 and decreased the number of apoptotic cells in HSV-1-infected IPEC-DQ cells, a porcine intestinal epithelial cell line. In contrast, A179L transfection of IPEC-DQ cells enhanced HSV-1-induced RIPK1, RIPK3, and MLKL phosphorylation and increased the number of necroptotic cells. Consistently, A179L also suppressed apoptosis but enhanced the necroptosis induced by two RNA viruses, Sendai virus (SeV) and influenza virus (IAV). Our study uncovers a previously unrecognized role of A179L in regulating cell death and suggests that A179L re-directs its anti-apoptotic activity to necroptosis.

scholarly journals Identification of a Continuously Stable and Commercially Available Cell Line for the Identification of Infectious African Swine Fever Virus in Clinical Samples

Viruses ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 820 ◽  
Author(s):  
Ayushi Rai ◽  
Sarah Pruitt ◽  
Elizabeth Ramirez-Medina ◽  
Elizabeth A. Vuono ◽  
Ediane Silva ◽  
...  
Keyword(s):  
Cell Line ◽  
Virus Isolation ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Clinical Samples ◽  
Outbreak Strain ◽  
Domestic Pigs ◽  
Current Outbreak ◽  
High Degree

African swine fever virus (ASFV) is causing outbreaks both in domestic pigs and wild boar in Europe and Asia. In 2018, the largest pig producing country, China, reported its first outbreak of African swine fever (ASF). Since then, the disease has quickly spread to all provinces in China and to other countries in southeast Asia, and most recently to India. Outbreaks of the disease occur in Europe as far west as Poland, and one isolated outbreak has been reported in Belgium. The current outbreak strain is highly contagious and can cause a high degree of lethality in domestic pigs, leading to widespread and costly losses to the industry. Currently, detection of infectious ASFV in field clinical samples requires accessibility to primary swine macrophage cultures, which are infrequently available in most regional veterinary diagnostic laboratories. Here, we report the identification of a commercially available cell line, MA-104, as a suitable substrate for virus isolation of African swine fever virus.

scholarly journals Crystal Structure of African Swine Fever Virus A179L with the Autophagy Regulator Beclin

Viruses ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 789 ◽  
Author(s):  
Suresh Banjara ◽  
Gareth L. Shimmon ◽  
Linda K. Dixon ◽  
Christopher L. Netherton ◽  
Mark G. Hinds ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Cell Death ◽  
Ligand Binding ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
B Cell Lymphoma ◽  
Vero Cells ◽  
Fever Virus ◽  
Site Directed Mutagenesis ◽  
Binding Groove

Subversion of programmed cell death-based host defence systems is a prominent feature of infections by large DNA viruses. African swine fever virus (ASFV) is a large DNA virus and sole member of the Asfarviridae family that harbours the B-cell lymphoma 2 or Bcl-2 homolog A179L. A179L has been shown to bind to a range of cell death-inducing host proteins, including pro-apoptotic Bcl-2 proteins as well as the autophagy regulator Beclin. Here we report the crystal structure of A179L bound to the Beclin BH3 motif. A179L engages Beclin using the same canonical ligand-binding groove that is utilized to bind to pro-apoptotic Bcl-2 proteins. The mode of binding of Beclin to A179L mirrors that of Beclin binding to human Bcl-2 and Bcl-xL as well as murine γ-herpesvirus 68. The introduction of bulky hydrophobic residues into the A179L ligand-binding groove via site-directed mutagenesis ablates binding of Beclin to A179L, leading to a loss of the ability of A179L to modulate autophagosome formation in Vero cells during starvation. Our findings provide a mechanistic understanding for the potent autophagy inhibitory activity of A179L and serve as a platform for more detailed investigations into the role of autophagy during ASFV infection.

scholarly journals Cytokine Storm in Domestic Pigs Induced by Infection of Virulent African Swine Fever Virus

2021 ◽  
Vol 7 ◽  
Author(s):  
Shuchao Wang ◽  
Jingyuan Zhang ◽  
Yanyan Zhang ◽  
Jinjin Yang ◽  
Lidong Wang ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Post Inoculation ◽  
Hemorrhagic Disease ◽  
Cytokine Storm ◽  
Domestic Pigs ◽  
Tnf Α ◽  
Anti Inflammatory

African swine fever, caused by African swine fever virus (ASFV), is a highly contagious hemorrhagic disease of domestic pigs. The current continent-wide pandemic has persisted for over 10 years, and its economy-devastating effect was highlighted after spreading to China, which possesses half of the world pig industry. So far, development of an effective and safe vaccine has not been finished largely due to the knowledge gaps in pathogenesis and immunology, particularly the role of cytokines in the host's immune response. Therefore, we performed experiments in domestic pigs to analyze the kinetics of representative circulating interferons (IFNs), interleukins (ILs), growth factors, tumor necrosis factors (TNFs), and chemokines induced by infection of type II virulent ASFV SY18. Pigs infected with this Chinese prototypical isolate developed severe clinical manifestations mostly from 3 days post inoculation (dpi) and died from 7 to 8 dpi. Serum analysis revealed a trend of robust and sustained elevation of pro-inflammatory cytokines including TNF-α, IFN-α, IL-1β, IL-6, IL-8, IL-12, IL-18, RANTES (regulated upon activation, normal T cell expressed and secreted), and IFN-γ-induced protein 10 (IP-10) from 3 dpi, but not the anti-inflammatory cytokines IL-10 and transforming growth factor-β (TGF-β). Moreover, secondary drastic increase of the levels of TNF-α, IL-1β, IL-6, and IL-8, as well as elevated IL-10, was observed at the terminal phase of infection. This pattern of cytokine secretion clearly drew an image of a typical cytokine storm characterized by delayed and dysregulated initiation of the secretion of pro-inflammatory cytokine and imbalanced pro- and anti-inflammatory response, which paved a way for further understanding of the molecular basis of ASFV pathogenesis.

scholarly journals Comparison of the Proteomes of Porcine Macrophages and a Stable Porcine Cell Line after Infection with African Swine Fever Virus

Viruses ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 2198
Author(s):  
Elisabeth Wöhnke ◽  
Walter Fuchs ◽  
Luise Hartmann ◽  
Ulrike Blohm ◽  
Sandra Blome ◽  
...  
Keyword(s):  
Cell Line ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Proteome Analysis ◽  
Viral Proteins ◽  
Viral Disease ◽  
Fever Virus ◽  
Specific Expression ◽  
Porcine Cell

African swine fever virus (ASFV), causing an OIE-notifiable viral disease of swine, is spreading over the Eurasian continent and threatening the global pig industry. Here, we conducted the first proteome analysis of ASFV-infected primary porcine monocyte-derived macrophages (moMΦ). In parallel to moMΦ isolated from different pigs, the stable porcine cell line WSL-R was infected with a recombinant of ASFV genotype IX strain “Kenya1033”. The outcome of the infections was compared via quantitative mass spectrometry (MS)-based proteome analysis. Major differences with respect to the expression of viral proteins or the host cell response were not observed. However, cell-specific expression of some individual viral proteins did occur. The observed modulations of the host proteome were mainly related to cell characteristics and function. Overall, we conclude that both infection models are suitable for use in the study of ASFV infection in vitro.

scholarly journals Development of Optimized Protocol for Culturing African Swine Fever Virus (ASFV) Field Isolates in MA104 Cells

2021 ◽  
Author(s):  
Hyeok-il Kwon ◽  
DUY tien DO ◽  
Hung Van Vo ◽  
Seung-Chul Lee ◽  
Min-Ho kim ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Vaccine Development ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Hemorrhagic Disease ◽  
Blood Samples ◽  
Ct Value ◽  
Tissue Homogenates

Abstract I. Background: ASFV causes a highly contagious hemorrhagic disease with a high mortality rates in domestic pigs. The virus has been isolated across various cell lines, but identifying a cell line to develop an effective commercial vaccine has been challenging which a major obstacle to effective vaccine development is identifying a commercial cell line that is suitable for high-yield viral replication.II. Methods and Results: The goal of this study was to identify a candidate commercial cell line for the replication of African swine fever virus (ASFV) by comparing several available cell lines with various medium factors. In the sensitivity test of cells, MA104 and MARC-145 had strong potential for ASFV replication. Next, MA104 cells were used to compare the adaptation of ASFV obtained from tissue homogenates and blood samples in various infectious media. At the 10th passage, the ASFV obtained from the blood sample had a significantly higher viral load than that obtained from the tissue sample (P = 0.000), exhibiting a mean Ct value = 20.39 ± 1.99 compared with 25.36 ± 2.11. For blood samples, ASFV grew on infectious medium B more robustly than on infectious medium A (P = 0.006), corresponding to a Ct value = 19.58 ± 2.10 versus 21.20 ± 1.47. ASFV originating from blood specimens continued to multiply gradually and peaked in the 15th passage, exhibiting a Ct value = 14.36 ± 0.22 in infectious medium B and a Ct value = 15.42 ± 0.14 in infectious medium A. However, there was no difference (P = 0.062) in ASFV growth between infectious media A and B when ASFV was cultured from tissue homogenates. III. Conclusions: A model was developed to enhance ASFV replication through adaptation to MA104 cells and the lack of mutation in serial culture passages may serve to maintain the immunogenicity of ASFV isolates when they are developed as vaccine candidates.

OBTAINING A STABLE CELL LINE EXPRESSING RECOMBINANT I329L PROTEIN OF AFRICAN SWINE FEVER VIRUS

2017 ◽  
Vol 52 (6) ◽  
pp. 1251-1258
Author(s):  
S.A. Katorkin ◽  
◽  
E.I. Katorkina ◽  
K.A. Mima ◽  
I.A. Titov ◽  
...  
Keyword(s):  
Cell Line ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Stable Cell Line

scholarly journals Crystal Structure of African Swine Fever Virus A179L with the Autophagy Regulator Beclin

2019 ◽  
Author(s):  
Suresh Banjara ◽  
Gareth Shimmon ◽  
Linda Dixon ◽  
Christopher Netherton ◽  
Mark Hinds ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Cell Death ◽  
Ligand Binding ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
B Cell Lymphoma ◽  
Vero Cells ◽  
Fever Virus ◽  
Site Directed Mutagenesis ◽  
Binding Groove

Subversion of programmed cell death-based host defence systems is a prominent feature of infections by large DNA viruses. African swine fever virus (ASFV) is a large DNA virus and sole member of the Asfarviridae family that harbors the B-cell lymphoma 2 or Bcl-2 homolog A179L. A179L has been shown to bind to a range of cell death inducing host proteins including pro-apoptotic Bcl-2 proteins as well as the autophagy regulator Beclin. Here we report the crystal structure of A179L bound to the Beclin BH3 motif. A179L engages Beclin using the same canonical ligand binding groove that is utilized to bind to pro-apoptotic Bcl-2 proteins. The mode of binding of Beclin to A179L mirrors that of Beclin binding to human Bcl-2 and Bcl-xL as well as murine gamma-herpesvirus 68. Introduction of bulky hydrophobic residues into the A179L ligand binding groove via site directed mutagenesis ablates binding of Beclin to A179L, leading to a loss of ability of A179L to modulate autophagosome formation in Vero cells during starvation. Our findings provide a mechanistic understanding for the potent autophagy inhibitory activity of A179L and serve as a platform for more detailed investigations into the role of autophagy during ASFV infection.

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