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ARS Home » Southeast Area » Athens, Georgia » U.S. National Poultry Research Center » Endemic Poultry Viral Diseases Research » Research » Publications at this Location » Publication #352454

Research Project: Intervention Strategies to Prevent and Control Enteric Diseases of Poultry

Location: Endemic Poultry Viral Diseases Research

Title: Generation of a recombinant Newcastle disease virus expressing two foreign genes for use as a multivalent vaccine and gene therapy vector

Author
item Hu, Haixia - Southwest University
item Roth, Jason - Boehringer Ingelheim Pharmaceuticals
item Yu, Qingzhong

Submitted to: Vaccine
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/23/2018
Publication Date: 6/30/2018
Citation: Hu, H., Roth, J.P., Yu, Q. 2018. Generation of a recombinant Newcastle disease virus expressing two foreign genes for use as a multivalent vaccine and gene therapy vector. Vaccine. 36:4846-4850. https://doi.org/10.1016/j.vaccine.2018.06.055.
DOI: https://doi.org/10.1016/j.vaccine.2018.06.055

Interpretive Summary: Many strains of Newcastle disease virus (NDV) have been developed as vaccine or gene therapy vectors during the past decade. However, most of these NDV vectors expressed only a single foreign gene as a monovalent vaccine or gene therapy agent. In the present study, we developed a novel NDV vector that can efficiently express two foreign genes for multivalent vaccine and gene therapy purposes. The red fluorescence protein (RFP) and the green fluorescence protein (GFP) were used as reporters for foreign gene expression. The RFP gene was inserted into the F gene as a second open reading frame, and the GFP was inserted between the F and HN genes as an independent transcription unit in a NDV LaSota strain-based infectious clone. A recombinant virus vectoring the RFP and GFP genes (rLS/IRES-RFP/GFP) was rescued from the recombinant infectious clone using reverse genetics technology. Biological assessments of rLS/IRES-RFP/GFP showed that it was slightly further attenuated in chickens, yet maintained similar growth abilities in cell cultures when compared to the parental LaSota virus. Expression of the RFP and GFP was detected from the rLS/IRES-RFP/GFP virus-infected cells by fluorescence microscopy. The data suggest that the rLS/IRES-RFP/GFP virus is safe and can be used as a vector for the development of multivalent vaccines and gene therapy agents.

Technical Abstract: Newcastle disease virus (NDV) has been used as a vector in the development of vaccines and gene therapy. A majority of these NDV vectors express only a single foreign gene through either an independent transcription unit (ITU) or an internal ribosomal entry site (IRES). In the present study, we combined the ITU and IRES methods to generate a novel NDV LaSota strain-based recombinant virus vectoring the red fluorescence protein (RFP) and the green fluorescence protein (GFP) genes. Biological assessments of the recombinant virus, rLS/IRES-RFP/GFP, showed that it was slightly attenuated in vivo, yet maintained similar growth dynamics and viral yields in vitro when compared to the parental LaSota virus. Expression of both the RFP and GFP was detected from the rLS/IRES-RFP/GFP virus-infected DF-1 cells by fluorescence microscopy. These data suggest that the rLS/IRES-RFP/GFP virus may be used as a multivalent vector for the development of vaccines and gene therapy agents.