Rescue of virulent class I Newcastle disease virus variant 9a5b-D5C1

Yu, Yong; Qiu, Xusheng; Xu, Dan; Zhan, Yuan; Chen, Meng; Wei, Nana; Chen, Hongjun; Tan, Lei; Yu, Shuang; Liu, Xiufan; Qin, Aijian; Ding, Chan

doi:10.1186/1743-422x-9-120

Cited by 13 publications

(7 citation statements)

References 34 publications

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“…To investigate whether over-expression of NDV V protein had an effect on STAT1 degradation, a series of plasmids were used [ 25 , 40 , 41 ]. The P gene ORF from LaSota, 9a5b and ZJ1 viruses was polymerase chain reaction (PCR)-amplified and ligated into the mammalian expression vector plasmid pCI-neo (Promega, USA).…”

Section: Resultsmentioning

confidence: 99%

Newcastle Disease Virus V Protein Targets Phosphorylated STAT1 to Block IFN-I Signaling

Qiu

Chen

et al. 2016

PLoS ONE

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Newcastle disease virus (NDV) V protein is considered as an effector for IFN antagonism, however, the mechanism remains unknown. In this study, the expression of STAT1 and phospho-STAT1 in cells infected with NDV or transfected with V protein-expressing plasmids were analyzed. Our results showed that NDV V protein targets phospho-STAT1 reduction in the cells depends on the stimulation of IFN-α. In addition, a V-deficient genotype VII recombinant NDV strain rZJ1-VS was constructed using reverse genetic technique to confirm the results. The rZJ1-VS lost the ability to reduce phospho-STAT1 and induced higher expression of IFN-responsive genes in infected cells. Furthermore, treatment with an ubiquitin E1 inhibitor PYR-41 demonstrated that phospho-STAT1 reduction was caused by degradation, but not de-phosphorylation. We conclude that NDV V protein targets phospho-STAT1 degradation to block IFN-α signaling, which adds novel knowledge to the strategies used by paramyxoviruses to evade IFN.

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Section: Resultsmentioning

confidence: 99%

Newcastle Disease Virus V Protein Targets Phosphorylated STAT1 to Block IFN-I Signaling

Qiu

Chen

et al. 2016

PLoS ONE

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“…Moreover, the NDV genome has to meet particular requirements for successful rescue such as the rule-of-six [31,32] and generation of the precise 3' and 5' ends [30,32]. To get the precise integral NDV genome, The traditional method is dividing the NDV genome into a set of short fragments from 6 to 11, and it takes months even years to complete for the multiple DNA fragments cloning [10,13,[33][34][35][36][37]. In this study, the genome of NDV was divided into three fragments 7bp, 5kb and 3kb.…”

Section: Discussionmentioning

confidence: 99%

Rescue of an Enterotropic Newcastle Disease Virus Strain ZM10 From Cloned cDNA and Stable Expressing an Inserted Foreign Gene

Wang

Zhao

et al. 2021

Preprint

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Background: Newcastle disease virus (NDV) strain ZM10, a typical enterotropic avirulent vaccine strain, has been widely used in in China for chickens against Newcastle disease. To elucidate its enterotropic mechanism and develop recombiant multivalent vaccines based on it, the reverse genetics system for NDV ZM10 is an indispensable platform.Results: A full-length cDNA clone of NDV ZM10 and three supporting plasmids were constructed using the ligation-independent cloning (LIC) method. Recombinant NDV rZM10 was successfully rescued after these plasmids were co-transfected into BHK-21 cells. Besides, the recombinant virus rZM10-RFP encoding the red fluorescent protein was generated by inserting the RFP gene into the full-length clone of NDV between the P and M genes. These rescued viruses were genetically and biologically identical to the parental strain and showed similar growth kinetics. Conclusion: The recovery system of NDV ZM10 strain was established, and can be used as a foundation for research on the enterotropic mechanism and development of multivalent vaccines against viral diseases of livestock and poultry.

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“…The BHK-21 cells were maintained in 6-well plates prior to transfection until 60–80% confluency. About 1 µg of full-length rAF-IL12 plasmid, 0.4 µg of pCI-neo-NP, 0.2 µg of pCI-neo-P and 0.2 µg of pCI-neo-L helper plasmids were transfected into the BHK-21 cell line using Lipofectamine™ 3000 27,36,37 . The culture supernatant was harvested at 24 hours post transfection and inoculated into 9-day old specific pathogen free (SPF) embryonated chicken eggs.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of a Recombinant Newcastle Disease Virus Expressing Human IL12 against Human Breast Cancer

Amin

Ani

Tan

et al. 2019

Sci Rep

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The Newcastle disease virus (NDV) strain AF2240 is an avian avulavirus that has been demonstrated to possess oncolytic activity against cancer cells. However, to illicit a greater anti-cancer immune response, it is believed that the incorporation of immunostimulatory genes such as IL12 into a recombinant NDV backbone will enhance its oncolytic effect. In this study, a newly developed recombinant NDV that expresses IL12 (rAF-IL12) was tested for its safety, stability and cytotoxicity. The stability of rAF-IL12 was maintained when passaged in specific pathogen free (SPF) chicken eggs from passage 1 to passage 10; with an HA titer of 29. Based on the results obtained from the MTT cytotoxic assay, rAF-IL12 was determined to be safe as it only induced cytotoxic effects against normal chicken cell lines and human breast cancer cells while sparing normal cells. Significant tumor growth inhibition (52%) was observed in the rAF-IL12-treated mice. The in vivo safety profile of rAF-IL12 was confirmed through histological observation and viral load titer assay. The concentration and presence of the expressed IL12 was quantified and verified via ELISA assay. In summary, rAF-IL12 was proven to be safe, selectively replicating in chicken and cancer cells and was able to maintain its stability throughout several passages; thus enhancing its potential as an anti-breast cancer vaccine.

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Rescue of virulent class I Newcastle disease virus variant 9a5b-D5C1

Cited by 13 publications

References 34 publications

Newcastle Disease Virus V Protein Targets Phosphorylated STAT1 to Block IFN-I Signaling

Newcastle Disease Virus V Protein Targets Phosphorylated STAT1 to Block IFN-I Signaling

Rescue of an Enterotropic Newcastle Disease Virus Strain ZM10 From Cloned cDNA and Stable Expressing an Inserted Foreign Gene

Evaluation of a Recombinant Newcastle Disease Virus Expressing Human IL12 against Human Breast Cancer

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