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© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.

Abstract

In this study, we developed a new recombinant virus rHVT-F using a Turkey herpesvirus (HVT) vector, expressing the fusion (F) protein of the genotype XII Newcastle disease virus (NDV) circulating in Peru. We evaluated the viral shedding and efficacy against the NDV genotype XII challenge in specific pathogen-free (SPF) chickens. The F protein expression cassette was inserted in the unique long (UL) UL45–UL46 intergenic locus of the HVT genome by utilizing a clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 gene-editing technology via a non-homologous end joining (NHEJ) repair pathway. The rHVT-F virus, which expressed the F protein stably in vitro and in vivo, showed similar growth kinetics to the wild-type HVT (wtHVT) virus. The F protein expression of the rHVT-F virus was detected by an indirect immunofluorescence assay (IFA), Western blotting, and a flow cytometry assay. The presence of an NDV-specific IgY antibody was detected in serum samples by an enzyme-linked immunosorbent assay (ELISA) in SPF chickens vaccinated with the rHVT-F virus. In the challenge experiment, the rHVT-F vaccine fully protects a high, and significantly reduced, virus shedding in oral at 5 days post-challenge (dpc). In conclusion, this new rHVT-F vaccine candidate is capable of fully protecting SPF chickens against the genotype XII challenge.

Details

Title
A Recombinant Turkey Herpesvirus Expressing the F Protein of Newcastle Disease Virus Genotype XII Generated by NHEJ-CRISPR/Cas9 and Cre-LoxP Systems Confers Protection against Genotype XII Challenge in Chickens
Author
Calderón, Katherine 1   VIAFID ORCID Logo  ; Rojas-Neyra, Aldo 2   VIAFID ORCID Logo  ; Carbajal-Lévano, Brigith 2   VIAFID ORCID Logo  ; Luján-Valenzuela, Luis 2 ; Ticona, Julio 2 ; Isasi-Rivas, Gisela 2 ; Montalvan, Angela 2 ; Criollo-Orozco, Manuel 2   VIAFID ORCID Logo  ; Huaccachi-Gonzáles, Edison 2 ; Tataje-Lavanda, Luis 2   VIAFID ORCID Logo  ; Alvarez, Karla Lucia F 2   VIAFID ORCID Logo  ; Fernández-Sánchez, Manolo 2 ; Fernández-Díaz, Manolo 2   VIAFID ORCID Logo  ; Tang, Na 3 ; Yao, Yongxiu 4 ; Nair, Venugopal 4   VIAFID ORCID Logo 

 Research and Development Laboratories, FARVET, Carretera Panamericana Sur N° 766 Km 198.5, Chincha Alta 11702, Peru; [email protected] (A.R.-N.); [email protected] (B.C.-L.); [email protected] (L.L.-V.); [email protected] (J.T.); [email protected] (G.I.-R.); [email protected] (A.M.); [email protected] (M.C.-O.); [email protected] (E.H.-G.); [email protected] (L.T.-L.); [email protected] (K.L.F.A.); [email protected] (M.F.-S.); [email protected] (M.F.-D.); Faculty of Medicine, Major National University of San Marcos, Av. Miguel Grau 755, Lima 15001, Peru 
 Research and Development Laboratories, FARVET, Carretera Panamericana Sur N° 766 Km 198.5, Chincha Alta 11702, Peru; [email protected] (A.R.-N.); [email protected] (B.C.-L.); [email protected] (L.L.-V.); [email protected] (J.T.); [email protected] (G.I.-R.); [email protected] (A.M.); [email protected] (M.C.-O.); [email protected] (E.H.-G.); [email protected] (L.T.-L.); [email protected] (K.L.F.A.); [email protected] (M.F.-S.); [email protected] (M.F.-D.) 
 Viral Oncogenesis Group & UK–China Centre of Excellence for Research on Avian Diseases, The Pirbright Institute, Pirbright, Surrey GU24 ONF, UK; [email protected] (N.T.); [email protected] (Y.Y.); [email protected] (V.N.); Binzhou Animal Science and Veterinary Medicine Academy & UK–China Centre of Excellence for Research on Avian Diseases, Binzhou 256600, China 
 Viral Oncogenesis Group & UK–China Centre of Excellence for Research on Avian Diseases, The Pirbright Institute, Pirbright, Surrey GU24 ONF, UK; [email protected] (N.T.); [email protected] (Y.Y.); [email protected] (V.N.) 
First page
793
Publication year
2022
Publication date
2022
Publisher
MDPI AG
e-ISSN
19994915
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2653042031
Copyright
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.