Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens.

Olga Chervyakova,Nurlan Sandybayev,Kunsulu Zakarya,Rinat A Maksyutov,Elmira Tailakova,Nurlan Kozhabergenov,Vitaliy Strochkov,Kulyaisan Sultankulova,Sandugash Sadikaliyeva

doi:10.3390/microorganisms9051005

Olga Chervyakova, Nurlan Sandybayev + Show 7 more

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https://doi.org/10.3390/microorganisms9051005

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Abstract

Capripoxviruses with a host range limited to ruminants have the great potential to be used as vaccine vectors. The aim of this work was to evaluate attenuated sheep pox virus (SPPV) vaccine strain NISKHI as a vector expressing several genes. Open reading frames SPPV020 (ribonucleotide kinase) and SPPV066 (thymidine kinase) were selected as sites for the insertion of foreign genes. Two integration plasmids with expression cassette were designed and constructed. Recombinant SPPVs expressing an enhanced green fluorescent protein (EGFP) (rSPPV(RRΔ)EGFP and rSPPV(TKΔ)EGFP), Foot-and-mouth disease virus capsid protein (VP1), and Brucella spp. outer membrane protein 25 (OMP25) (rSPPV(RRΔ)VP1A-(TKΔ)OMP25) were generated under the transient dominant selection method. The insertion of foreign genes into the SPPV020 and SPPV066 open reading frames did not influence the replication of the recombinant viruses in the cells. Successful foreign gene expression in vitro was assessed by luminescent microscopy (EGFP) and Western blot (VP1 and OMP25). Our results have shown that foreign genes were expressed by rSPPV both in permissive (lamb testicles) and non-permissive (bovine kidney, saiga kidney, porcine kidney) cells. Mice immunized with rSPPV(RRΔ)VP1A-(TKΔ)OMP25 elicited specific antibodies to both SPPV and foreign genes VP1 and OMP25. Thus, SPPV NISKHI may be used as a potential safe immunogenic viral vector for the development of polyvalent vaccines.

Highlights

Introduction published maps and institutional affilThe use of poxviruses as vectors for foreign gene expression was first shown in 1982 by using the vaccinia virus [1]
It was shown that the knockout of the ribonucleotide reductase small subunit gene does not affect the replication of capripoxviruses and can be used to insert foreign sequences [14]
To evaluate the efficacy of sheep pox virus (SPPV) NISKHI as a recombinant vaccine vector, we investigated the immunogenicity of rSPPV(RR-)VP1A-(TK-)outer membrane protein 25 (OMP25) in mice

Summary

Introduction

The use of poxviruses as vectors for foreign gene expression was first shown in 1982 by using the vaccinia virus [1]. Since a great number of vaccinia virus strains were used in the expression of genes of various pathogens [3,4]. A number of recombinant vaccinia viruses expressing many genes of animal viruses have been developed [7,8]. Capripoxviruses were used as vectors to express foreign genes for vaccine development [9,10,11]. The genus Capripoxvirus includes three members: lumpy skin disease virus, sheep pox virus (SPPV) and goat pox virus.

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