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

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.

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Anti-Influenza Protective Efficacy of a H6 Virus-Like Particle in Chickens

Vaccines ◽

10.3390/vaccines8030465 ◽

2020 ◽

Vol 8 (3) ◽

pp. 465 ◽

Cited By ~ 1

Author(s):

Wan-Zhen Zhu ◽

Yi-Chi Wen ◽

Shu-Yi Lin ◽

Ting-Chih Chen ◽

Hui-Wen Chen

Keyword(s):

Matrix Protein ◽

Protective Efficacy ◽

Expression System ◽

Influenza Viruses ◽

Vaccine Antigen ◽

Effective Vaccine ◽

Particle Structure ◽

Species Barrier ◽

Virus Like Particle

H6 avian influenza viruses (AIVs) have a worldwide distribution, and they pose a potential concern for public health. In Taiwan, H6 AIVs have circulated in domestic chickens for more than 40 years, and certain strains have crossed the species barrier to infect mammals. With the goal of containing the disease, there is a pressing need to develop a safe and effective vaccine for pandemic preparedness. In this study, we prepared a virus-like particle (VLP) that consisted of the hemagglutinin (HA) and matrix protein 1 (M1) derived from a H6 AIV as a vaccine antigen, and we examined the immunogenicity and protective efficacy when combined with an adjuvant in a chicken model. Full-length HA and M1 protein genes were cloned and expressed using a baculovirus expression system, and VLPs were purified from the supernatant of insect cell cultures. We performed nanoparticle-tracking analysis and transmission electron microscopy to validate that the particle structure and properties resembled the native virions. In animal experiments, specific-pathogen-free chickens that received the H6 VLPs in combination with an adjuvant showed superior H6N1 virus-specific serum IgG and hemagglutination-inhibition antibody responses, which lasted more than 112 days. Following the H6N1 viral challenge, the vaccinated chickens showed reduced viral replication in the lungs, kidneys and conjunctival/cloacal shedding. The antibodies induced in the chickens by the vaccine were able to cross-react with the H6N1 human isolate and drifted avian H6N1 isolates. In summary, the H6 VLP vaccine elicited superb immunogenicity in vivo, and the use of an adjuvant further enhanced the antiviral protective efficacy. This vaccine formulation could potentially be used to manage H6 influenza virus infections in chickens.

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Essential Domains for Ribonucleoprotein Complex Formation Required for Retrotransposition of Telomere-Specific Non-Long Terminal Repeat Retrotransposon SART1

Molecular and Cellular Biology ◽

10.1128/mcb.00096-06 ◽

2006 ◽

Vol 26 (13) ◽

pp. 5168-5179 ◽

Cited By ~ 20

Author(s):

Takumi Matsumoto ◽

Mitsuhiro Hamada ◽

Mizuko Osanai ◽

Haruhiko Fujiwara

Keyword(s):

Long Terminal Repeat ◽

Expression System ◽

Baculovirus Expression System ◽

Open Reading Frames ◽

Terminal Repeat ◽

Ltr Retrotransposons ◽

C Terminus ◽

Functional Features

ABSTRACT Non-long terminal repeat (LTR) retrotransposons are major components of the higher eukaryotic genome. Most of them have two open reading frames (ORFs): ORF2 encodes mainly the endonuclease and reverse transcriptase domains, but the functional features of ORF1 remain largely unknown. We used telomere-specific non-LTR retrotransposon SART1 in Bombyx mori and clarified essential roles of the ORF1 protein (ORF1p) in ribonucleoprotein (RNP) formation by novel approaches: in vitro reconstitution and in vivo/in vitro retrotransposition assays using the baculovirus expression system. Detailed mutation analyses showed that each of the three CCHC motifs at the ORF1 C terminus are essential for SART1 retrotransposition and are involved in packaging the SART1 mRNA specifically into RNP. We also demonstrated that amino acid residues 555 to 567 and 285 to 567 in the SART1 ORF1p are crucial for the ORF1p-ORF1p and ORF1p-ORF2p interactions, respectively. The loss of these domains abolishes protein-protein interaction, leading to SART1 retrotransposition deficiency. These data suggest that systematic formation of RNP composed of ORF1p, ORF2p, and mRNA is mainly mediated by ORF1p domains and is a common, essential step for many non-LTR retrotransposons encoding the two ORFs.

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Human β nerve growth factor obtained from a baculovirus expression system has potent in vitro and in vivo neurotrophic activity

Experimental Neurology ◽

10.1016/0014-4886(90)90047-v ◽

1990 ◽

Vol 110 (1) ◽

pp. 11-24 ◽

Cited By ~ 32

Author(s):

Jim Barnett ◽

Preston Baecker ◽

Carol Routledge-Ward ◽

Hela Bursztyn-Pettegrew ◽

Joan Chow ◽

...

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Expression System ◽

Baculovirus Expression System ◽

Nerve Growth ◽

Neurotrophic Activity ◽

Baculovirus Expression

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Protective efficacy of crude virus-like particle vaccine against HPAI H5N1 in chickens and its application on DIVA strategy

Influenza and Other Respiratory Viruses ◽

10.1111/j.1750-2659.2012.00396.x ◽

2012 ◽

Vol 7 (3) ◽

pp. 340-348 ◽

Cited By ~ 10

Author(s):

Jae-Keun Park ◽

Dong-Hun Lee ◽

Ha-Na Youn ◽

Myeong-Seob Kim ◽

Yu-Na Lee ◽

...

Keyword(s):

Protective Efficacy ◽

Virus Like Particle ◽

Hpai H5n1

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In vivo calcineurin crystals formed using the baculovirus expression system

Microscopy Research and Technique ◽

10.1002/(sici)1097-0029(19960501)34:1<77::aid-jemt11>3.0.co;2-m ◽

1996 ◽

Vol 34 (1) ◽

pp. 77-86 ◽

Cited By ~ 15

Author(s):

G.Y. Fan ◽

Fausto Maldonado ◽

Y. Zhang ◽

Randall Kincaid ◽

Mark H. Ellisman ◽

...

Keyword(s):

Expression System ◽

Baculovirus Expression System ◽

Baculovirus Expression

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Pigs Immunized with the Virus-like Particle Vaccine Are Protected against the Hepatitis E-3 Virus

Vaccines ◽

10.3390/vaccines9111265 ◽

2021 ◽

Vol 9 (11) ◽

pp. 1265

Author(s):

Hyeon-Jeong Go ◽

Byung-Joo Park ◽

Hee-Seop Ahn ◽

Dong-Hwi Kim ◽

Da-Yoon Kim ◽

...

Keyword(s):

Expression System ◽

Vaccine Dose ◽

Baculovirus Expression System ◽

Hepatitis E ◽

Positive Control ◽

Negative Control ◽

Fecal Shedding ◽

Viral Shedding ◽

Baculovirus Expression ◽

Virus Like Particle

In this study, we generated the HEV virus-like particle (VLP) vaccine expressing 239 amino acids (367–605 aa) of the HEV-3 ORF2 using the baculovirus expression system. The HEV-3-239-VLP vaccine efficacy was evaluated by dividing 12 pathogen-free pigs into four groups: negative control, positive control, 100 μg VLP-, and 200 μg VLP-vaccinated groups for 10 weeks. The pigs in either of the vaccinated groups were administered the corresponding first and booster doses on weeks 0 and 2. At week 4, the positive control and two vaccinated groups were challenged with 106 HEV-3 genomic equivalent copies; viremia and fecal shedding of the virus were identified in pigs in the positive control and 100 μg VLP-vaccinated pigs showed transient viremia and fecal viral shedding. However, no viruses were detected in the serum or fecal samples of the 200 μg VLP-vaccinated pigs. The 100 and 200 μg VLP-vaccinated pigs had significantly higher (p < 0.01) anti-HEV antibodies than the negative control pigs from weeks 6–10 with normal levels of liver enzymes. The 200 μg VLP-vaccinated pigs showed statistically less liver tissue fibrosis (p < 0.05) than that of the positive control pigs. Thus, the novel baculovirus expression system-generated VLP vaccine dose-dependently protects against HEV-3 challenge and may be useful in other animal species, including humans.

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Tyrosine Phosphorylation of the Proto-Oncoprotein Raf-1 Is Regulated by Raf-1 Itself and the Phosphatase Cdc25A

Molecular and Cellular Biology ◽

10.1128/mcb.19.7.4819 ◽

1999 ◽

Vol 19 (7) ◽

pp. 4819-4824 ◽

Cited By ~ 36

Author(s):

Kai Xia ◽

Robert S. Lee ◽

Radha P. Narsimhan ◽

Nishit K. Mukhopadhyay ◽

Benjamin G. Neel ◽

...

Keyword(s):

Tyrosine Phosphorylation ◽

Expression System ◽

Baculovirus Expression System ◽

Wild Type ◽

Phosphatase Inhibitors ◽

System A ◽

New Findings ◽

Nih 3T3 ◽

Tyrosine Phosphate

ABSTRACT There is a growing body of evidence demonstrating that Raf-1 is phosphorylated on tyrosines upon stimulation of a variety of receptors. Although detection of Raf-1 tyrosine phosphorylation has remained elusive, genetic analyses have demonstrated it to be important for Raf-1 activation. Here we report new findings which indicate that Raf-1 tyrosine phosphorylation is regulated in vivo. In both a mammalian and baculovirus expression system, a kinase-inactive allele of Raf-1 was found to be tyrosine phosphorylated at levels much greater than that of wild-type Raf-1. The level of tyrosine phosphate on Raf-1 was markedly increased upon treatment with phosphatase inhibitors either before or after cell lysis. Cdc25A was found to dephosphorylate Raf-1 on tyrosines that resulted in a significant decrease in Raf-1 kinase activity. In NIH 3T3 cells, coexpression of wild-type Raf-1 and phosphatase-inactive Cdc25A led to a marked increase in Raf-1 tyrosine phosphorylation in response to platelet-derived growth factor. These data suggest that the tyrosine phosphorylation of Raf-1 is regulated not only by itself but also by Cdc25A.

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The potential application of Trichoplusia ni granulovirus En3 enhancin as a synergist in baculovirus-based insecticides

10.48035/tesis/2454/41034 ◽

2021 ◽

Author(s):

◽

Adriana Ricarte Bermejo

Keyword(s):

Mass Production ◽

Trichoplusia Ni ◽

Expression System ◽

Baculovirus Expression System ◽

Active Matter ◽

Expression Systems ◽

Lepidopteran Pests ◽

Baculovirus Expression ◽

Present Thesis

The increased costs associated with baculovirus mass-production urge the search for synergistic products that reduce the amount of active matter. In the present thesis, a synergistic factor with great potential for baculovirus-based formulations was expressed and produced using a baculovirus expression system. The main achievement of the present thesis is that the in vivo production of solubilized enhancins using baculovirus-based expression systems can be used to improve the efficacy of biological insecticides against lepidopteran pests, reducing the active matter of bioinsecticides and making them commercially competitive with chemicals.

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

Viruses ◽

10.3390/v13122393 ◽

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.

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