In silico prediction and in vitro identification of bluetongue virus 4 VP5 protein B-cell epitopes

Abstract

VP5, the outer capsid protein of bluetongue virus (BTV), plays an important role in viral penetration and antibody-mediated viral neutralization. Therefore, VP5 represents an important target for development of vaccines and diagnostic tests. In this study, we use bioinformatic tools to predict nine antigenic B cell epitopes in the VP5 protein of a BTV serotype 4 (BTV4) isolate from China. Further, we generate five BTV4 VP5-specific monoclonal antibodies (MAbs) and define their corresponding epitopes using a set of VP5-derived peptides expressed as maltose-binding protein (MBP) fusion proteins. The five identified epitopes map to amino acids 119-134, 257-272, 286-301, 322-337, and 481-496 of the VP5 protein. Importantly, the epitopes identified using VP5-derived peptides do not correlate with our bioinformatic prediction of antibody epitopes. Identification and characterization of BTV4 VP5 protein epitopes may aid the development of diagnostic tools and provide information with which to study the structure of the BTV VP5 protein.

Immuno-informatics Study Identifies Conserved T Cell Epitopes in Non-structural Proteins of Bluetongue Virus Serotypes: Formulation of Computationally Optimized Next-Generation Broad-spectrum Multiepitope Vaccine

“…Based on the molecular interaction patterns, the complex of mouse vaccine and TLR3 was found to have the second highest number of hydrogen bonds after mouse vaccine-TLR4 complex and 276 non-bonded interactions. The outcome of molecular docking is consistent with the docking results of the previous studies (Bhattacharya and Roy 2008, Sun, Sun et al 2014, Russell, Parbhoo et al 2018).…”

Analysis of Conserved, Computationally Predicted Epitope Regions for VP5 and VP7 Across three Orbiviruses

“…On VP5, the linear B-cell epitope 265-LSGID-269 67 is highly conserved across all 3 viruses; however, the 266-S amino acid was not conserved in AHS. Other B-cell epitopes have previously been identified for VP5 68 – 70 but have been found to not be effective at providing protection across serotypes; this could potentially be due to the lack of amino acid sequence conservation not only between the different orbiviruses but even also between the serotypes.…”

Identification of three antigen epitopes on the nucleocapsid protein of the genotype C of bovine parainfluenza virus type 3

“…Antigenic determinants, especially conserved epitopes, should be taken into consideration when choosing monoclonal antibodies (mAbs) for diagnostic purposes. Moreover, defining conserved epitopes can suggest targets for potential pharmacological inhibitors and inform vaccine design (Liu et al, 2013;Sun et al, 2014). Therefore, the current study was aimed at characterizing the antigenic determinants of BPIV3c NP using mAbs and three epitopes had been identified with mAbs against a series of overlapping recombinant BPIV3c NP fragments expressed in Escherichia coli.…”