Recombinant Sheep Pox Virus Proteins Elicit Neutralizing Antibodies.

Olga Chervyakova,Lev Nemchinov,Rosemarie Hammond,Daniyar Beisenov,Valentin Zaitsev,Gulshan Stanbekova,Natalia Kovalskaya,Abylay Sansyzbay,Nurlan Sandybayev,Muratbay Mambetaliyev,Elmira Tailakova,Bulat Iskakov,Kulyaisan Sultankulova,Yergali Abduraimov,Vitaliy Strochkov

doi:10.3390/v8060159

Abstract

The aim of this work was to evaluate the immunogenicity and neutralizing activity of sheep pox virus (SPPV; genus Capripoxvirus, family Poxviridae) structural proteins as candidate subunit vaccines to control sheep pox disease. SPPV structural proteins were identified by sequence homology with proteins of vaccinia virus (VACV) strain Copenhagen. Four SPPV proteins (SPPV-ORF 060, SPPV-ORF 095, SPPV-ORF 117, and SPPV-ORF 122), orthologs of immunodominant L1, A4, A27, and A33 VACV proteins, respectively, were produced in Escherichia coli. Western blot analysis revealed the antigenic and immunogenic properties of SPPV-060, SPPV-095, SPPV-117 and SPPV-122 proteins when injected with adjuvant into experimental rabbits. Virus-neutralizing activity against SPPV in lamb kidney cell culture was detected for polyclonal antisera raised to SPPV-060, SPPV-117, and SPPV-122 proteins. To our knowledge, this is the first report demonstrating the virus-neutralizing activities of antisera raised to SPPV-060, SPPV-117, and SPPV-122 proteins.

Highlights

East and Central Asia, India, China, and Central and Northern Africa
Genes sheep pox virus (SPPV)-060 (GenBank ID: NP_659632), SPPV-095 (GenBank ID: NP_659667), SPPV-117 (GenBank ID: NP_659689), and SPPV-122 (GenBank ID: NP_659694) were PCR-amplified from viral DNA, extracted from SPPV virions with Trizol (Invitrogen, Carlsbad, CA, USA), using the primer pairs described in Table 1
Induction of recombinant protein expression in bacteria using IPTG resulted in the production of SPPV-060 (22 kDa; Figure 2A, lane 3), SPPV-095 (20 kDa; Figure 2C, lane 6), SPPV-117 (19 kDa; Figure 2B, lane 2), and SPPV-122 (16 kDa; Figure 2C, lane 2) proteins

Summary

Introduction

The disease causes significant economic losses by reduced hide and wool quality, the prevention of the importation of new breeds of sheep into endemic regions, and other production losses in animal husbandry worldwide [1,2,3,4]. The agent of sheep pox disease is sheep pox virus (SPPV), the type species of the Capripoxvirus genus, subfamily Chordopoxvirinae, family Poxviridae. In addition to SPPV, the capripoxviruses include goatpox virus (GTPV) and lumpy skin disease virus, all of which cause pox skin lesions, are of great economic importance, and are potentially emerging disease threats [5]. To control the spread of sheep pox infection, attenuated virus vaccines are widely used in the Republic of Kazakhstan and other Former. In spite of the high protection afforded by live vaccines, the possible presence of residual virulence in the latter restricts their use in disease-free areas and has led to the exploration of new approaches in vaccine development involving recombinant DNA technology [9,10,11,12,13,14,15,16,17]

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