Conformation of the VP2 protein of bluetongue virus (BTV) determines the involvement in virus neutralization of highly conserved epitopes within the BTV serogroup

Bluetongue (BT) is a midge-borne OIE-notifiable disease of ruminants caused by the bluetongue virus (BTV). There are at least 29 BTV serotypes as determined by serum neutralization tests and genetic analyses of genome segment 2 encoding serotype immunodominant VP2 protein. Large parts of the world are endemic for multiple serotypes. The most effective control measure of BT is vaccination. Conventionally live-attenuated and inactivated BT vaccines are available but have their specific pros and cons and are not DIVA compatible. The prototype Disabled Infectious Single Animal (DISA)/DIVA vaccine based on knockout of NS3/NS3a protein of live-attenuated BTV, shortly named DISA8, fulfills all criteria for modern veterinary vaccines of sheep. Recently, DISA8 with an internal in-frame deletion of 72 amino acid codons in NS3/NS3a showed a similar ideal vaccine profile in cattle. Here, the DISA/DIVA vaccine platform was applied for other serotypes, and pentavalent DISA/DIVA vaccine for “European” serotypes 1, 2, 3, 4, 8 was studied in sheep and cattle. Protection was demonstrated for two serotypes, and neutralization Ab titers indicate protection against other included serotypes. The DISA/DIVA vaccine platform is flexible in use and generates monovalent and multivalent DISA vaccines to combat specific field situations with respect to Bluetongue.

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Identification of a novel bluetongue virus 1-specific B-cell epitope using a monoclonal antibody against the VP2 protein

Archives of Virology ◽

10.1007/s00705-012-1590-6 ◽

2013 ◽

Vol 158 (5) ◽

pp. 1099-1104 ◽

Cited By ~ 4

Author(s):

P. Wei ◽

E. C. Sun ◽

N. H. Liu ◽

T. Yang ◽

Q. Y. Xu ◽

...

Keyword(s):

Monoclonal Antibody ◽

B Cell ◽

Bluetongue Virus ◽

Cell Epitope ◽

Vp2 Protein ◽

B Cell Epitope

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The Tip Region on VP2 Protein of Bluetongue Virus Contains Potential IL-4-Inducing Amino Acid Peptide Segments

Pathogens ◽

10.3390/pathogens10010003 ◽

2020 ◽

Vol 10 (1) ◽

pp. 3

Author(s):

Jia-Ling Yang ◽

Chia-Yi Chang ◽

Chih-Shuan Sheng ◽

Chia-Chi Wang ◽

Fun-In Wang

Keyword(s):

Innate Immunity ◽

Allergic Reaction ◽

Synthetic Peptides ◽

Bluetongue Virus ◽

Messenger Rna ◽

Nonstructural Protein ◽

Interleukin 4 ◽

Hemorrhagic Disease ◽

Vp2 Protein ◽

Peptide Segments

Bluetongue is an infectious viral hemorrhagic disease of domestic and wild ruminants that has a considerable economic impact on domestic ruminants. There are currently at least 29 serotypes of bluetongue virus (BTV) in the world. Noteworthily, the pathogenesis among BTV serotypes is different, even in the same animal species. In this study, BTV2/KM/2003 and BTV12/PT/2003 were used to investigate the differential immunological effects on bovine peripheral blood mononuclear cells (PBMCs). The BTV viral load and the expression of cytokine messenger RNA (mRNA) in PBMCs were measured by fluorescence-based real-time reverse-transcription PCR (qRT-PCR). The immunofluorescence assay (IFA) was applied to detect BTV signals in monocyte-derived macrophages (MDMs). The SWISS-MODEL and IL-4pred prediction tools were used to predict the interleukin 4 (IL-4)-inducing peptides in BTV-coat protein VP2. Synthetic peptides of VP2 were used to stimulate PBMCs for IL-4-inducing capability. This study demonstrated that the cytokine profiles of BTV-induced PBMCs were significantly different between BTV2/KM/2003 and BTV12/PT/2003. BTV2 preferentially activated the T helper 2 (Th2) pathway, represented by the early induction of IL-4, and likely fed back to inhibit the innate immunity. In contrast, BTV12 preferentially activated the innate immunity, represented by the induction of tumor necrosis factor -α (TNF-α) and interleukin 1 (IL-1), with only minimal subsequent IL-4. The BTV nonstructural protein 3 antibody (anti-BTV-NS3) fluorescent signals demonstrated that monocytes in PBMCs and MDMs were the preferred targets of BTV replication. Bioinformatics analysis revealed that the capability to induce IL-4 was attributed to the tip region of the VP2 protein, wherein a higher number of predicted peptide segments on BTVs were positively correlated with the allergic reaction reported in cattle. Synthetic peptides of BTV2-VP2 induced significant IL-4 within 12–24 h post-infection (hpi) in PBMCs, whereas those of BTV12 did not, consistent with the bioinformatics prediction. Bovine PBMCs and synthetic peptides together seem to serve as a good model for pursuing the BTV-induced IL-4 activity that precedes the development of an allergic reaction, although further optimization of the protocol is warranted.

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Host-dependent variation of Bluetongue virus neutralization

Journal of Invertebrate Pathology ◽

10.1016/0022-2011(91)90038-r ◽

1991 ◽

Vol 57 (1) ◽

pp. 37-42 ◽

Cited By ~ 1

Author(s):

Franziska B. Grieder ◽

Kevin T. Schultz

Keyword(s):

Bluetongue Virus ◽

Virus Neutralization

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Identification of a novel bluetongue virus 1 specific B cell epitope using monoclonal antibodies against the VP2 protein

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2021.05.053 ◽

2021 ◽

Author(s):

Aiping Wang ◽

Jinran Du ◽

Hua Feng ◽

Jingming Zhou ◽

Yumei Chen ◽

...

Keyword(s):

Monoclonal Antibodies ◽

B Cell ◽

Bluetongue Virus ◽

Cell Epitope ◽

Vp2 Protein ◽

B Cell Epitope

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Computer-Aided Structure Prediction of Bluetongue Virus Coat Protein VP2 Assisted by Optimized Potential for Liquid Simulations (OPLS)

Current Topics in Medicinal Chemistry ◽

10.2174/1568026620666200516153753 ◽

2020 ◽

Vol 20 (19) ◽

pp. 1720-1732 ◽

Cited By ~ 1

Author(s):

Leena Prajapati ◽

Ravina Khandelwal ◽

Kadapakkam Nandabalan Yogalakshmi ◽

Anjana Munshi ◽

Anuraj Nayarisseri

Keyword(s):

Active Site ◽

Viral Protein ◽

Bluetongue Virus ◽

3D Structure ◽

Protein Domain ◽

Dynamics Simulation ◽

Local Alignment ◽

Structure Comparison ◽

Vp2 Protein ◽

The Stability

Background: The capsid coated protein of Bluetongue virus (BTV) VP2 is responsible for BTV transmission by the Culicoides vector to vertebrate hosts. Besides, VP2 is responsible for BTV entry into permissive cells and hence plays a major role in disease progression. However, its mechanism of action is still unknown. Objective: The present investigation aimed to predict the 3D structure of Viral Protein 2 of the bluetongue virus assisted by Optimized Potential for Liquid Simulations (OPLS), structure validation, and an active site prediction. Methods: The 3D structure of the VP2 protein was built using a Python-based Computational algorithm. The templates were identified using Smith waterman’s Local alignment. The VP2 protein structure validated using PROCHECK. Molecular Dynamics Simulation (MDS) studies were performed using an academic software Desmond, Schrodinger dynamics, for determining the stability of a model protein. The Ligand-Binding site was predicted by structure comparison using homology search and proteinprotein network analysis to reveal their stability and inhibition mechanism, followed by the active site identification. Results: The secondary structure of the VP2 reveals that the protein contains 220 alpha helix atoms, 40 310 helix, 151 beta sheets, 134 coils and 424 turns, whereas the 3D structure of Viral Protein 2 of BTV has been found to have 15774 total atoms in the structure. However, 961 amino acids were found in the final model. The dynamical cross-correlation matrix (DCCM) analysis tool identifies putative protein domains and also confirms the stability of the predicted model and their dynamical behavior difference with the correlative fluctuations in motion. Conclusion: The biological interpretation of the Viral Protein 2 was carried out. DCCM maps were calculated, using a different coordinate reference frame, through which, protein domain boundaries and protein domain residue constituents were identified. The obtained model shows good reliability. Moreover, we anticipated that this research should play a promising role in the identification of novel candidates with the target protein to inhibit their functional significance.

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Group-specific and Type-specific Gel Diffusion Precipitin Tests for Bluetongue Virus Serotype 20 and Related Viruses

Australian Journal of Biological Sciences ◽

10.1071/bi9880553 ◽

1988 ◽

Vol 41 (4) ◽

pp. 553 ◽

Cited By ~ 1

Author(s):

JM Sharp ◽

I R Littlejohns ◽

T D St George

Keyword(s):

Cell Cultures ◽

Virus Type ◽

Experimental Infection ◽

Bluetongue Virus ◽

Neutralization Test ◽

Virus Neutralization Test ◽

Virus Neutralization ◽

Specific Test ◽

Virus Serotype ◽

Gel Diffusion

Using antigens prepared from cell cultures infected by bluetongue (BLU) virus type 20 (BLU-20), and sera from cattle which had recovered from experimental infection by that virus, two distinct precipitin reactions were demonstrated by immunodiffusion. Two distinct gel diffusion precipitin tests were developed based on these reactions. The antigen of one was common to BLU-20 and two other Australian BLU isolates, CSIRO 154 (BLU-21) and CSIRO 156 (BLU-l). It was therefore concluded to be a group-specific test. The antigen of the second appeared to be unique to BLU-20. The test based on this antigen correlated well with the virus neutralization test for BLU-20 and it was therefore concluded to be type-specific.

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