Identification of four novel group-specific bluetongue virus NS3 protein B-cell epitopes
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RESEARCH
Open Access
Identification of four novel group-specific bluetongue virus NS3 protein B-cell epitopes Zhang Qin†, Sun EnCheng†, Xu QingYuan, Yang Tao, Wang HaiXiu, Feng YuFei, Li JunPing, Lv Shuang and Wu DongLai*
Abstract Background: The non-structural protein 3 (NS3) of bluetongue virus (BTV) is the second smaller non-structural protein produced in host cells, playing an important role in BTV trafficking and release. Results: In this study, we generated five BTV NS3-reactive monoclonal antibodies (mAbs), named 3D8, 2G9, 1B5, 4H8, and 2B12. A panel of overlapping NS3-derived peptides representing the entirety of the BTV15 NS3 protein was screened to identify linear peptide epitopes recognized by each mAb. Based on the initial screen, a series of progressively truncated peptides were produced to identify the minimal linear peptide sequence required to maintain mAb binding. We found that mAb 3D8 reacted with the motif 36PPRYA40, 2G9 reacted with the motif 82AEAFRDDVRLRQIK95, 1B5 reacted with the motif 205YNDAVRMSF213, 2B12 and 4H8 reacted with the motif 204SYNDAVRMSF213. Sequence alignments demonstrated that these linear epitopes are highly conserved among all BTV serotypes, consistent with the observation that each mAb was able to recognize cells infected with BTV1-24 serotypes tested and each identified B cell epitope was able to be recognized by BTV-infect sheep serum. Conclusion: This collection of mAbs along with defined linear epitopes may provide useful reagents for investigations of NS3 protein function and the development of BTV group-specific diagnostics. Keywords: Bluetongue virus, NS3 protein, Group-specific monoclonal antibody, B-cell epitopes
Background Bluetongue disease (BT) is a hemorrhagic disease of ruminants caused by bluetongue virus (BTV) which is spread through the bite of midges of the genus Culicoides [1–3]. BT is primarily found in the tropics, subtropics and temperate zones due to the limited distribution of Culicoides midges and presents a threat to the development of livestock farming [4–6]. Due to the severe impact of BT, the Office International Des Epizooties (OIE) lists BT as a notifiable disease. BTV is the prototype member of the genus Orbivirus within the Reoviridae family. The BTV genome consists of 10 double-stranded RNA segments differ in lengths that encode seven structural proteins (VP1-VP7), and four nonstructural protein NS1, NS2, NS3/NS3a and NS4 [7]. The BTV genome is contained in a double layer capsid. The * Correspondence: [email protected] † Equal contributors The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, People’s Republic of China
outer virion capsid is composed of VP2 and VP5 protein, and accounts for approximately 40 % of the total protein content. The inner capsid consists of VP3 and VP7, and three secondary proteins including VP1, VP4 and VP6. Antigenic differences in the VP2 account
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