scholarly journals Detection of African Swine Fever Virus-Specific Antibodies Using the Recombinant Viral Proteins p32 and p54 Expressed in Insect Cells

2006 ◽  
Vol 40 (3) ◽  
pp. 271-276
Author(s):  
Kenichi SAKAMOTO ◽  
Makoto YAMAKAWA ◽  
Tomoyuki TSUDA ◽  
Yosuke MURAKAMI
Keyword(s):  
Insect Cells ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Viral Proteins ◽  
Fever Virus ◽  
Specific Antibodies

scholarly journals EFFECTS OF MODERATELY VIRULENT AFRICAN SWINE FEVER VIRUS ON INTERLEUKIN-10 PRODUCTION

2019 ◽  
pp. 23-28 ◽  
Author(s):  
A. S. Pershin ◽  
I. V. Shevchenko ◽  
A. S. Igolkin ◽  
Ye. V. Aronova ◽  
N. N. Vlasova
Keyword(s):  
Immune Response ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Noticeable Effect ◽  
Specific Antibodies ◽  
Virulent Virus ◽  
Virus Isolates ◽  
Highly Virulent

A characteristic feature of African swine fever virus (ASFV) is the ability to escape from host immune response, affecting macrophages and replicating in them. Besides, ASFV - specific antibodies do not completely neutralize the virus. Cytokines are important factors for various viral infection pathologies. The virulence of ASFV isolates may depend on the capacity to regulate cytokine expression by macrophages. Thus, when comparing in vitro and in vivo cytokine production by macrophages, it was established that infection with low virulent virus isolates leads to an immune response with a predominance of cytokines involved in cellular immunity, such as INF-α and IL-12p40, as compared with infection with highly virulent isolates. The aim of this paper was to study the effect of African swine fever virus on the production of IL-10, a pleiotropic cytokine that inhibits synthesis of cytokines and shows a strong antiinflammatory effect. For this, 12 piglets were experimentally infected intramuscularly with a continuous cell culture-adapted ASFV isolate Vero25 at a dose of 10 HAdU per animal followed by control infection of surviving animals with the reference virus isolate Arm 07 at a dose of 1,000 HAdU per animal. Temperature measurements were taken and blood sampling to obtain serum was conducted during the experiment. IL-10 amount in blood sera was determined using Invitrogen test systems (Thermo Fisher, USA). A higher IL-10 level (15.8–173 pg/ml) was observed in blood sera of dead animals infected with a moderately virulent virus, as compared with surviving pigs (4–5 pg/ml). No correlation between the speed of appearance of specific antibodies and IL-10 serum levels has been established. No noticeable effect of the IL-10 serum level prior to infection on the survival rate of animals has been observed. Further studies are needed to establish a causal relationship, including study of the expression of various cytokines during infection with both low- and highly virulent virus isolates.

scholarly journals Antigenic Properties and Diagnostic Potential of African Swine Fever Virus Protein pp62 Expressed in Insect Cells

2006 ◽  
Vol 44 (3) ◽  
pp. 950-956 ◽  
Author(s):  
C. Gallardo ◽  
E. Blanco ◽  
J. M. Rodriguez ◽  
A. L. Carrascosa ◽  
J. M. Sanchez-Vizcaino
Keyword(s):  
Insect Cells ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Virus Protein ◽  
Diagnostic Potential ◽  
Antigenic Properties

scholarly journals Immunization of Pigs with Recombinant Plasmids Containing Genes of Ubiquitinated p30, p54 and CD2v Proteins of African Swine Fever Virus

2020 ◽  
Vol 70 (1) ◽  
pp. 92-109
Author(s):  
Almaz R Imatdinov ◽  
Anna S Kazakova ◽  
Milanko Šekler ◽  
Diana Yu Morozova ◽  
Valentina M Lyska ◽  
...  
Keyword(s):  
Cytotoxic T Lymphocytes ◽  
Cellular Immunity ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Chimeric Proteins ◽  
Cellular Mechanisms ◽  
Specific Antibodies ◽  
Human Ubiquitin ◽  
Protective Proteins

AbstractThree recombinant plasmid constructs, expressing chimeric proteins containing human ubiquitin fused to an ectodomain of one of the potentially protective proteins (p30, p54 and CD2v) of the attenuated MK-200 strain of African swine fever virus (ASFV), were created as potential inductors of specific antiviral cellular immunity. Three-time immunization of pigs with the mixture of these plasmids led to the formation of virus-specific cytotoxic T-lymphocytes (CTL), but did not induce production of virus-specific antibodies. After challenge with the homologous parental virulent ASFV strain M-78 at a dose of 103 HAD50, all five animals (four immunized pigs and one naïve) fell between the 4th and 7th days post infection. The obtained results demonstrated that induction of CTL did not protect pigs against challenge with the virulent ASFV. Balanced activation of CTL and antibody-mediated cellular mechanisms should be investigated.

scholarly journals African Swine Fever Virus A528R Inhibits TLR8 Mediated NF-κB Activity by Targeting p65 Activation and Nuclear Translocation

Viruses ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2046
Author(s):  
Xueliang Liu ◽  
Da Ao ◽  
Sen Jiang ◽  
Nengwen Xia ◽  
Yulin Xu ◽  
...  
Keyword(s):  
Promoter Activity ◽  
Nuclear Translocation ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Viral Proteins ◽  
Fever Virus ◽  
Economic Losses ◽  
Effective Vaccine ◽  
Hemorrhagic Disease ◽  
Host Innate Immunity

African swine fever (ASF) is mainly an acute hemorrhagic disease which is highly contagious and lethal to domestic pigs and wild boars. The global pig industry has suffered significant economic losses due to the lack of an effective vaccine and treatment. The African swine fever virus (ASFV) has a large genome of 170–190 kb, encoding more than 150 proteins. During infection, ASFV evades host innate immunity via multiple viral proteins. A528R is a very important member of the polygene family of ASFV, which was shown to inhibit IFN-β production by targeting NF-κB, but its mechanism is not clear. This study has shown that A528R can suppress the TLR8-NF-κB signaling pathway, including the inhibition of downstream promoter activity, NF-κB p65 phosphorylation and nuclear translocation, and the antiviral and antibacterial activity. Further, we found the cellular co-localization and interaction between A528R and p65, and ANK repeat domains of A528R and RHD of p65 are involved in their interaction and the inhibition of p65 activity. Therefore, we conclude that A528R inhibits TLR8-NF-κB signaling by targeting p65 activation and nuclear translocation.

scholarly journals Functionality and Cell Anchorage Dependence of the African Swine Fever Virus Gene A179L, a Viralbcl-2 Homolog, in Insect Cells

1998 ◽  
Vol 72 (12) ◽  
pp. 10227-10233 ◽  
Author(s):  
Alejandro Brun ◽  
Fernando Rodríguez ◽  
José M. Escribano ◽  
Covadonga Alonso
Keyword(s):  
Cell Survival ◽  
Intracellular Signaling ◽  
Insect Cells ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Virus Gene ◽  
Mammalian Cell Lines ◽  
Extracellular Matrix Components ◽  
Baculovirus Infection

ABSTRACT The African swine fever virus gene A179L has been shown to be a functional member of the ced9/bcl-2 family of apoptosis inhibitors in mammalian cell lines. In this work we have expressed the A179L gene product (p21) under the control of the baculovirus polyhedrin promoter using a baculovirus system. Expression of the A179L gene neither altered the baculovirus replication phenotype nor delayed the shutoff of cellular protein synthesis, but it extended the survival of the infected insect cells to very late times postinfection. The increase in cell survival rates correlated with a marked apoptosis reduction after baculovirus infection. Interestingly, prevention of apoptosis was observed when recombinant baculovirus infections were carried out in monolayer cell cultures but not when cells were infected in suspension, suggesting a cell anchorage dependence for p21 function in insect cells. Cell survival was enhanced under optimal conditions of cell attachment and cell-to-cell contact as provided by extracellular matrix components or poly-d-lysine. Since it was observed that cytoskeleton organization varied depending on culture conditions of insect cells (grown in monolayer versus grown in suspension), these results suggested that A179L might regulate apoptosis in insect cells only when the cytoskeletal support of intracellular signaling is maintained upon cell adhesion. Thus, cell shape and cytoskeleton status might allow variations in intracellular transduction of signals related to cell survival in virus-infected cells.

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.

A comparison of two indirect immune electron microscopy procedures using monoclonal antibody to localize specific African swine fever virus (ASFV) proteins in infected cells and virus

1983 ◽  
Vol 41 ◽  
pp. 800-801
Author(s):  
G. J. Letchworth III ◽  
T. C. Whyard ◽  
S. H. Wool
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Grid Method ◽  
Viral Proteins ◽  
Fever Virus ◽  
Immune Electron Microscopy ◽  
Infected Cells

The primary purpose of this study was to devise a procedure in which a large number of monoclonal antibodies could be tested against several African swine fever virus (ASFV) preparations (virus, infected cells). The procedure had to be rapid, use a small sample and provide for some TEM.Monoclonal antibodies were produced by standard methods using myeloma cells and lymphocytes from mice immunized against ASF viral proteins. Antibodies were characterized by immunoprecipitation of radio-labelled viral proteins (Fig 1).The viral preparations used in this study were either infected macrophages, culture fluid supernatant from ASFV-infected macrophages or red blood cells from infected pigs. Each of the three viral preparations was tested by two indirect immune electron microscopy (IEM) methods; a grid method and a test tube method. In the grid method, formvar coated grids were floated on a viral preparation then for one hour each on the mouse monoclonal antibody and finally on ferritin tagged sheep anti-mouse IgG with washes between each step.

scholarly journals Systematic analysis of longitudinal serological responses of pigs infected experimentally with African swine fever virus

2007 ◽  
Vol 88 (9) ◽  
pp. 2426-2434 ◽  
Author(s):  
Ana Luísa Reis ◽  
R. M. E. Parkhouse ◽  
Ana Raquel Penedos ◽  
Carlos Martins ◽  
Alexandre Leitão
Keyword(s):  
Immune Response ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Viral Proteins ◽  
Fever Virus ◽  
Antibody Responses ◽  
Igg Antibody ◽  
Clear Trend ◽  
Systematic Analysis ◽  
Antibody Titres

The protective immune response to African swine fever virus (ASFV) includes both cellular and serological components. In this study, the role of antibodies in the pathogenicity and diagnosis of African swine fever (ASF) was explored. Accordingly, total and Ig isotype antibody responses against the 12 viral proteins previously demonstrated to be the main targets of serological immunity were evaluated in longitudinally collected sera from pigs infected experimentally with the non-pathogenic ASFV/NH/P68 isolate. Strong total IgG antibody responses were observed against viral proteins E183L/p54, K205R/‘unassigned’, A104R/histone-like and B602L/‘unassigned’; therefore, IgM, IgG1 and IgG2 responses to these proteins were also determined. One protein stimulating IgM (K205R) may have practical potential for the detection of recently infected animals. There was a clear trend towards an IgG1 response to all of the proteins. This may reflect a dominant Th2-controlled immune response. In order to identify possible correlations between these serological responses and the pathogenesis of ASF, total IgG responses to the 12 recombinant proteins were compared in asymptomatic and chronically infected animals. For the proteins NP419L/DNA ligase, CP312R, B646L/p73, K196R/thymidine kinase and K205R, the antibody titres were significantly higher in animals developing lesions. One exception was the antibody response to the A104R/histone-like protein, which was higher in asymptomatic than in chronically infected pigs, suggesting that antibodies against this protein might be an indicator of an effective immune response or that this response is somehow involved in protection.

Genetic markers of the African swine fever virus

2020 ◽  
Vol 23 (04) ◽  
pp. 21-26
Author(s):  
A.K. Sibgatullova ◽  
◽  
M.E. Vlasov ◽  
I.A. Titov ◽  
◽  
...  
Keyword(s):  
Genetic Markers ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus
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