scholarly journals Characterization of Ictalurid herpesvirus 1 Glycoprotein ORF59 and Its Potential Role on Virus Entry into the Host Cells

Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2393
Author(s):  
Shu-Xin Li ◽  
Fei Yu ◽  
Hong-Xun Chen ◽  
Xiao-Dong Zhang ◽  
Li-Hui Meng ◽  
...  
Keyword(s):  
Channel Catfish ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Inhibitory Effect ◽  
Infection Process ◽  
Host Cells ◽  
Economic Losses ◽  
Ovary Cells ◽  
Channel Catfish Virus ◽  
Herpesvirus 1

The channel catfish virus (CCV, Ictalurid herpesvirus 1) has caused sustained economic losses in the fish industry because of its strong infectivity and pathogenicity. Thus, it is necessary to determine the function of viral proteins in the CCV infection process. The present study aimed to characterize CCV glycoprotein ORF59 and explore its impact on virus infection in host cells. Firstly, its exclusive presence in the membrane fraction of the cell lysate and subcellular localization verified that CCV ORF59 is a viral membrane protein expressed at late-stage infection. A protein blocking assay using purified His6 tagged ORF59, expressed in sf9 insect cells using a baculovirus expression system, indicated a dose-dependent inhibitory effect of recombinant ORF59 protein on virus invasion. Knockdown of the ORF59 using a short hairpin (shRNA) showed that ORF59 silencing decreased the production of infectious virus particles in channel catfish ovary cells. The results of this study suggest that recombinant ORF59 protein might inhibit CCV entry into the host cells. These findings will promote future studies of the key functions of glycoprotein ORF59 during CCV infection.

scholarly journals Evaluation of Protective Efficacy of Influenza Virus Like Particles Prepared from H5N1 Virus of Clade 2.2.1.2 in Chickens

Vaccines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 715
Author(s):  
Mohamed H. El-Husseiny ◽  
Naglaa M. Hagag ◽  
Peter Pushko ◽  
Irina Tretyakova ◽  
Mahmoud M. Naguib ◽  
...  
Keyword(s):  
H5n1 Virus ◽  
Protective Efficacy ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Economic Losses ◽  
Protection Level ◽  
Virus Like Particle ◽  
Specific Pathogen ◽  
Hpai H5n1

Highly pathogenic Avian Influenza (HPAI) viruses continue to cause severe economic losses in poultry species worldwide. HPAI virus of subtype H5N1 was reported in Egypt in 2006, and despite vaccination efforts, the virus has become endemic. The current study aims to evaluate the efficacy of a virus-like particle (VLP) based vaccine in vivo using specific pathogen-free (SPF) chickens. The vaccine was prepared from the HPAI H5N1 virus of clade 2.2.1.2 using the baculovirus expression system. The VLPs were quantitated and characterized, including electron microscopy. In addition, the protection level of the VLPs was evaluated by using two different regimens, including one dose and two-dose vaccinated groups, which gave up to 70% and 100% protection level, respectively. The results of this study emphasize the potential usefulness of the VLPs-based vaccine as an alternative vaccine candidate for the control of AIV infection in poultry.

Acyclovir inhibits channel catfish virus replication and protects channel catfish ovary cells from apoptosis

2021 ◽  
Vol 292 ◽  
pp. 198249
Author(s):  
Kai Hao ◽  
Sheng Yuan ◽  
Fei Yu ◽  
Xiao Hui Chen ◽  
Wen Ji Bian ◽  
...  
Keyword(s):  
Virus Replication ◽  
Channel Catfish ◽  
Ovary Cells ◽  
Channel Catfish Virus

Purification of Channel Catfish virus, a fish herpesvirus

1978 ◽  
Vol 24 (11) ◽  
pp. 1335-1338 ◽  
Author(s):  
Jean Robin ◽  
Alice Rodrigue
Keyword(s):  
Electron Microscopy ◽  
Channel Catfish ◽  
Host Cells ◽  
Host Proteins ◽  
Peg 6000 ◽  
Channel Catfish Virus ◽  
Viral Particles ◽  
Isopycnic Centrifugation

Channel Catfish virus was highly purified from host cells by precipitation with PEG-6000 and isopycnic centrifugation in a metrizamide gradient. As calculated from reconstruction experiments, only 0.09 and 0.05% respectively of the host DNA and the host proteins were recovered at the position of the viral band. The final recovery of infectivity was about 30%. Electron microscopy showed mostly intact viral particles.

scholarly journals Characterizing the Type 6 Secretion System (T6SS) and its role in the virulence of avian pathogenic Escherichia coli strain APECO18

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12631
Author(s):  
Aline L. de Oliveira ◽  
Nicolle L. Barbieri ◽  
Darby M. Newman ◽  
Meaghan M. Young ◽  
Lisa K. Nolan ◽  
...  
Keyword(s):  
Culture Media ◽  
Coli Strain ◽  
Infection Process ◽  
Host Cells ◽  
Economic Losses ◽  
Poultry Production ◽  
E Coli ◽  
Intestinal Infections ◽  
Animal Pathogens ◽  
Bacterial Replication

Avian pathogenic E. coli is the causative agent of extra-intestinal infections in birds known as colibacillosis, which can manifest as localized or systemic infections. The disease affects all stages of poultry production, resulting in economic losses that occur due to morbidity, carcass condemnation and increased mortality of the birds. APEC strains have a diverse virulence trait repertoire, which includes virulence factors involved in adherence to and invasion of the host cells, serum resistance factors, and toxins. However, the pathogenesis of APEC infections remains to be fully elucidated. The Type 6 secretion (T6SS) system has recently gained attention due to its role in the infection process and protection of bacteria from host defenses in human and animal pathogens. Previous work has shown that T6SS components are involved in the adherence to and invasion of host cells, as well as in the formation of biofilm, and intramacrophage bacterial replication. Here, we analyzed the frequency of T6SS genes hcp, impK, evpB, vasK and icmF in a collection of APEC strains and their potential role in virulence-associated phenotypes of APECO18. The T6SS genes were found to be significantly more prevalent in APEC than in fecal E. coli isolates from healthy birds. Expression of T6SS genes was analyzed in culture media and upon contact with host cells. Mutants were generated for hcp, impK, evpB, and icmF and characterized for their impact on virulence-associated phenotypes, including adherence to and invasion of host model cells, and resistance to predation by Dictyostelium discoideum. Deletion of the aforementioned genes did not significantly affect adherence and invasion capabilities of APECO18. Deletion of hcp reduced resistance of APECO18 to predation by D. discoideum, suggesting that T6SS is involved in the virulence of APECO18.

scholarly journals Preparation of Chicken Anemia Virus (CAV) Virus-Like Particles and Chicken Interleukin-12 for Vaccine Development Using a Baculovirus Expression System

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 262
Author(s):  
Ta-Yuan Tseng ◽  
Yee-Chen Liu ◽  
Yu-Chen Hsu ◽  
Poa-Chun Chang ◽  
Ming-Kun Hsieh ◽  
...  
Keyword(s):  
Vaccine Development ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Chicken Anemia Virus ◽  
Economic Losses ◽  
Interleukin 12 ◽  
Cell Mediated Immunity ◽  
Single Chain ◽  
Virus Like Particles ◽  
Baculovirus Expression

Chicken infectious anemia (CIA) is a poultry disease that causes huge economic losses in the poultry industry worldwide. Commercially available CIA vaccines are derived from wild-type chicken anemia viruses (CAVs) by serial passage in cells or chicken embryos. However, these vaccinal viruses are not completely attenuated; therefore, they can be transmitted vertically and horizontally, and may induce clinical symptoms in young birds. In this study, we sought to eliminate these issues by developing a subunit vaccine exploiting the CAV structural proteins, engineering recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells that contained both the viral protein 1 (VP1) and VP2 of CAV. Moreover, we produced single-chain chicken interleukin-12 (chIL-12) in the same system, to serve as an adjuvant. The recombinant VP1 was recognized by chicken anti-CAV polyclonal antibodies in Western blotting and immunofluorescence assays, and the bioactivity of the recombinant chIL-12 was confirmed by stimulating interferon-γ (IFN-γ) secretion in chicken splenocytes. Furthermore, the ability of the recombinant VP1 to generate self-assembling virus-like particles (VLPs) was confirmed by transmission electron microscopy. Specific pathogen-free (SPF) chickens inoculated with VLPs and co-administered the recombinant chIL-12 induced high CAV-specific antibodies and cell-mediated immunity. Taken together, the VLPs produced by the baculovirus expression system have the potential to be a safe and effective CIA vaccine. Finally, we demonstrated the utility of recombinant chIL-12 as an adjuvant for poultry vaccine development.

Biochemical Chgaracterization of Recombinant Baculovirus 373 K/E p53 Protein

2018 ◽  
Vol 9 (03) ◽  
pp. 20204-20223
Author(s):  
Maghsoudi, Hossein ◽  
U Pati
Keyword(s):  
Dna Binding ◽  
P53 Protein ◽  
Expression System ◽  
Gel Filtration ◽  
Recombinant Baculovirus ◽  
Baculovirus Expression System ◽  
Cross Linking ◽  
Mobility Shift ◽  
Dna Complexes ◽  
P53 Antibodies

In this study, we expressed and purified the recombinant baculovirus 373 K/E p53 protein in a baculovirus expression system to characterize this mutant and compare it with wild type p53. Gel- filtration chromatography and chemical cross-linking experiments indicated that purified recombinant baculovirus 373 K/E p53 protein assembles into multimeric forms ranging from tetramers to polymers. Gel-mobility shift assays and protein-DNA cross-linking studies demonstrated that the recombinant protein binds, to a consensus DNA target as a dimer but that additional p53 mutant molecules may then associate with the preformed p53-dimer-DNA complexes to form a larger p53_DNA complexes. These observations suggest that the p53 mutant tetramers and polymers that forms the minimal p53 mutant complex in solution dissociated upon DNA binding to form p53 mutant dimmer DNA complexes. The DNA binding activity of this mutant was then investigated using electrophoretic mobility shift assays as well as supershift assay with anti-p53 antibodies. Binding of the anti-p53 antibody PAb421to the oligomerization promoting domain on p53 stimulated the sequential formation of both the p53_dimer DNA and larger p53-DNA complexes

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