Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens
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 SP...
Main Authors: | , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2021-05-01
|
Series: | Microorganisms |
Subjects: | |
Online Access: | https://www.mdpi.com/2076-2607/9/5/1005 |
id |
doaj-6ae051718fa24485b33502ae8cf1e41e |
---|---|
record_format |
Article |
spelling |
doaj-6ae051718fa24485b33502ae8cf1e41e2021-05-31T23:24:48ZengMDPI AGMicroorganisms2076-26072021-05-0191005100510.3390/microorganisms9051005Engineering of Recombinant Sheep Pox Viruses Expressing Foreign AntigensOlga Chervyakova0Elmira Tailakova1Nurlan Kozhabergenov2Sandugash Sadikaliyeva3Kulyaisan Sultankulova4Kunsulu Zakarya5Rinat A. Maksyutov6Vitaliy Strochkov7Nurlan Sandybayev8Research Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanState Research Center of Virology and Biotechnology “Vector”, Koltsovo, 630559 Novosibirsk Region, RussiaResearch Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, RK ME&S–Science Committee, Gvardeiskiy 080409, KazakhstanCapripoxviruses 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 <i>Brucella</i> 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.https://www.mdpi.com/2076-2607/9/5/1005SPPVvaccine vectorintegration plasmidsthymidine kinaseribonucleotide reductase |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Olga Chervyakova Elmira Tailakova Nurlan Kozhabergenov Sandugash Sadikaliyeva Kulyaisan Sultankulova Kunsulu Zakarya Rinat A. Maksyutov Vitaliy Strochkov Nurlan Sandybayev |
spellingShingle |
Olga Chervyakova Elmira Tailakova Nurlan Kozhabergenov Sandugash Sadikaliyeva Kulyaisan Sultankulova Kunsulu Zakarya Rinat A. Maksyutov Vitaliy Strochkov Nurlan Sandybayev Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens Microorganisms SPPV vaccine vector integration plasmids thymidine kinase ribonucleotide reductase |
author_facet |
Olga Chervyakova Elmira Tailakova Nurlan Kozhabergenov Sandugash Sadikaliyeva Kulyaisan Sultankulova Kunsulu Zakarya Rinat A. Maksyutov Vitaliy Strochkov Nurlan Sandybayev |
author_sort |
Olga Chervyakova |
title |
Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens |
title_short |
Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens |
title_full |
Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens |
title_fullStr |
Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens |
title_full_unstemmed |
Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens |
title_sort |
engineering of recombinant sheep pox viruses expressing foreign antigens |
publisher |
MDPI AG |
series |
Microorganisms |
issn |
2076-2607 |
publishDate |
2021-05-01 |
description |
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 <i>Brucella</i> 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. |
topic |
SPPV vaccine vector integration plasmids thymidine kinase ribonucleotide reductase |
url |
https://www.mdpi.com/2076-2607/9/5/1005 |
work_keys_str_mv |
AT olgachervyakova engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT elmiratailakova engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT nurlankozhabergenov engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT sandugashsadikaliyeva engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT kulyaisansultankulova engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT kunsuluzakarya engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT rinatamaksyutov engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT vitaliystrochkov engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens AT nurlansandybayev engineeringofrecombinantsheeppoxvirusesexpressingforeignantigens |
_version_ |
1721417617301831680 |