Reverse Genetics of Newcastle Disease Virus

Cardenas-Garcia, Stivalis; Afonso, Claudio L.

doi:10.1007/978-1-4939-6964-7_10

Cited by 12 publications

(9 citation statements)

References 43 publications

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“…The PCR product was then digested with Sac II and Spe I restriction enzymes and the 1561 bp fragment was then introduced back to the same site in the genomic cDNA clone pNDV/AI4 for constructing the plasmid pO/AI4. Recovery of the recombinant NDV using pO/AI4 was performed as described previously [ 14 , 15 , 16 ], and the rescued virus was designated as NDV O/AI4. The biologic characterization of the rescued viruses was evaluated by standard determined the mean death time (MDT), the intracerebral pathogenicity index (ICPI), and the intravenous pathogenicity index (IVPI) [ 19 ].…”

Section: Methodsmentioning

confidence: 99%

“…The objective of this study is to investigate the effects of the antigenic difference in the HN protein between the vaccine strain and the challenge strain on the protection efficiency in terms of the virulent virus load and shedding. Toward this end, using reverse genetics we generated a vaccine strain designated as NDV O/AI4 by replacing the HN gene of the vaccine strain NDV/AI4 with that of the variant NDV strain JS-14-12-Ch bearing the E347K and G362A co-mutation, while NDV/AI4 was attenuated from the NDV JS5/05 strain [ 14 , 15 , 16 , 17 ]. We compared the immune efficiency difference between LaSota, AI4 and NDV O/AI4 against the challenge strain JS-14-12-Ch, while the antibody titers of the vaccinated birds with each of the three vaccines were at the same level.…”

Section: Introductionmentioning

confidence: 99%

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Effects of the HN Antigenic Difference between the Vaccine Strain and the Challenge Strain of Newcastle Disease Virus on Virus Shedding and Transmission

Liu

Jun

et al. 2017

Viruses

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Newcastle disease (ND) leading to heavy economic losses to the poultry industry worldwide is caused by Newcastle disease virus (NDV). Even though intensive vaccination programs have been implemented in many countries, virulent NDV can still be frequently isolated in well-vaccinated flocks. We compared the protection efficiency of LaSota and two sub-genotype VIId vaccines, NDV/AI4 and NDV O/AI4, in which NDV O/AI4 was constructed by replacing the hemagglutinin–neuraminidase (HN) gene of the vaccine strain NDV/AI4 with that from the variant NDV strain JS-14-12-Ch by the cross hemagglutination inhibition test and immune protection test. The number of birds shedding the virus and the titer of the shedding virus from the challenged birds were tested to evaluate the protection efficiency in the immune protection test. The cross hemagglutination inhibition and neutralization tests between JS-14-12-Ch and the three vaccines displayed a significant antigenic difference between JS-14-12-Ch and LaSota or NDV/AI4, but not between JS-14-12-Ch and NDV O/AI4. The results of the immune protection test showed that NDV O/AI4 could provide improved protection as determined by a significant decrease in both the number of birds shedding the virus and the titer of the shedding virus from the challenged birds. The results in this study indicated that the antigenic similarity between the vaccine strain and the challenge strain is important in reducing the shedding of virulent virus in which the congruence of the NDV HN protein may play a critical role.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effects of the HN Antigenic Difference between the Vaccine Strain and the Challenge Strain of Newcastle Disease Virus on Virus Shedding and Transmission

Liu

Jun

et al. 2017

Viruses

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“…The technology of the reverse genetic system (RGS) for NDV was established in Europe two decades ago, which has been successfully applied to research on pathogenicity, replication regulatory mechanisms, and vaccine vectors [33]. Meanwhile, the LaSota strain has been used as a safe and effective live vaccine that protects against NDV challenge for several decades, because it can elicit both humoral and cell-mediated immune responses, and vaccine production is cost-effective.…”

Section: Introductionmentioning

confidence: 99%

A Recombinant La Sota Vaccine Strain Expressing Multiple Epitopes of Infectious Bronchitis Virus (IBV) Protects Specific Pathogen-Free (SPF) Chickens against IBV and NDV Challenges

et al. 2019

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Infectious bronchitis (IB) and Newcastle disease (ND) are two major infectious diseases that are a threat to the domestic poultry industry. In this study, we successfully generated a recombinant LaSota candidate vaccine strain, rNDV-IBV-T/B, which expresses a short, synthetic, previously identified IBV S1 multi-epitope cassette using the reverse genetic system. The recombinant virus was propagated in nine-day-old embryonated chicken eggs for 20 passages and genetic stability was confirmed by whole genome DNA sequencing. The recombinant virus had a hemagglutination (HA) titer of 2 10 , mean death time (MDT) of 118 hours, and intracerebral pathogenicity index (ICPI) of 0.05. None of these were significantly different from the parental Newcastle disease virus (NDV) LaSota strain (p > 0.05). Vaccination of white leghorn chickens at one day of age with 10 6 EID 50 rNDV-IBV-T/B provided 90% protection against virulent IBV M41 challenge at three weeks of age, which was significantly higher than the protection of the control group vaccinated with phosphate-buffered saline (PBS) (p < 0.05). The ciliostasis scores of rNDV-IBV-T/B-vaccinated and LaSota-vaccinated groups were 4.2 and 37.6, respectively, which indicated that rNDV-IBV-T/B vaccination reduced the pathogenicity of IBV toward the trachea. Furthermore, real-time RT-PCR assay showed that the rNDV-IBV-T/B vaccination resulted in low levels of viral load (647.80 ± 49.65 RNA copies) in the trachea four days post-challenge, which is significantly lower than groups vaccinated with PBS (8591.25 ± 311.10 RNA copies) or LaSota (7742.60 ± 298.50 RNA copies) (p < 0.05). Meanwhile, the same dose of rNDV-IBV-T/B vaccination provided complete protection against velogenic NDV F48E9 challenge. These results demonstrate that the rNDV-IBV-T/B strain is a promising vaccine candidate to control both IB and ND simultaneously. Furthermore, epitope-based live vector vaccines provide an alternative strategy for the development of cost-effective and, broadly, cross-protective vaccines.

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“…For an overview of the NDV purification procedure, see Figure 1 . Note that although there are published protocols available for rescuing recombinant NDV (rNDV), 21 , 22 one of the aims of this paper was to provide a comprehensive protocol for both the production and the purification of high-titer NDV. Because oncolytic NDV is designated as a Risk Group 2 pathogen in Canada, the procedures described in this methods paper were conducted in a BSL-2 laboratory within a type IIA biological safety cabinet.…”

Section: Methodsmentioning

confidence: 99%

Production and Purification of High-Titer Newcastle Disease Virus for Use in Preclinical Mouse Models of Cancer

Santry

McAusland

Susta

et al. 2018

Molecular Therapy - Methods & Clinical Development

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Newcastle disease virus (NDV) is a single-stranded, negative-sense RNA virus in the Paramyxoviridae family. Although primarily an avian pathogen, NDV is a potent oncolytic virus that has been shown to be safe and effective in a variety of preclinical cancer models and human clinical trials. To produce virus for oncolytic trials, NDV is commonly amplified in embryonated chicken eggs and purified from the allantoic fluid. Conventional methods for purifying virus from allantoic fluid often result in relatively low-titer preparations containing high levels of impurities, including immunogenic chicken host cell proteins from allantoic fluid. However, large quantities of virus need to be delivered intravenously to administer oncolytic NDV systemically to mice. This route of administration requires virus preparations that are both highly concentrated (to enable delivery of small volumes) and highly pure (to limit toxic effects from contaminants). Given the accumulation of promising preclinical and clinical data demonstrating the efficacy of NDV as an oncolytic agent, strategies for increasing the titer and purity of NDV preparations are sorely needed to allow for effective intravenous administration in mice. Here, we describe an optimized protocol for the rescue, production, and purification of high-titer in vivo-grade NDV for preclinical studies in mouse models.

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Reverse Genetics of Newcastle Disease Virus

Cited by 12 publications

References 43 publications

Effects of the HN Antigenic Difference between the Vaccine Strain and the Challenge Strain of Newcastle Disease Virus on Virus Shedding and Transmission

Effects of the HN Antigenic Difference between the Vaccine Strain and the Challenge Strain of Newcastle Disease Virus on Virus Shedding and Transmission

A Recombinant La Sota Vaccine Strain Expressing Multiple Epitopes of Infectious Bronchitis Virus (IBV) Protects Specific Pathogen-Free (SPF) Chickens against IBV and NDV Challenges

Production and Purification of High-Titer Newcastle Disease Virus for Use in Preclinical Mouse Models of Cancer

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