Comprehensive immunoinformatics approach for developing a multi-epitope subunit vaccine against lumpy skin disease

Résumé fourni par la source

• The study proposes a multi-epitope vaccine (MEV) against Lumpy Skin Disease Virus (LSDV) using immunoinformatics tools, addressing the inefficacy and limitations of existing commercial vaccines. • Conserved and immunodominant CTL, HTL, and B-cell epitopes from LSDV proteins (GPCR, p32, RPO30) were identified and validated for antigenicity, non-allergenicity, and non-toxicity. • The designed MEV construct showed high binding affinity and stable interaction with the bovine TLR4 receptor, as confirmed by molecular docking and molecular dynamics (MD) simulation studies. • Structural modeling and refinement revealed a stable, high-quality tertiary structure, while in silico cloning into the pET28a(+) vector ensured its potential for scalable recombinant vaccine production. • The LSDV-MEV candidate has the potential to trigger broad humoral and cellular immune responses, supporting its further validation for effective LSDV prophylaxis . Lumpy skin disease virus (LSDV) is an emerging transboundary pathogen associated with lumpy skin disease (LSD). Recent outbreaks of LSDV have now been reported from previously unaffected regions, including the Indian subcontinent. Rapid transmission with high morbidity and mortality, enormously impacts the bovine species, resulting in devastating economic consequences on the livestock sector. Therefore, it necessitates expedited, appropriate attention to LSDV prophylaxis and control. Various LSDV suboptimal vaccines are prevailing that range in effectiveness, efficacy, safety, and side effects. Therefore, by utilizing the immunoinformatics approach mainly, a multi-epitope vaccine candidate was designed. To achieve this, conserved regions of three key LSDV proteins GPCR, P32, and RPO30 were first identified from complete Indian LSDV genomes. From these conserved segments, epitope mapping was performed, resulting in the selection of 16 high-affinity cytotoxic T lymphocyte (CTL) epitopes, 14 helper T lymphocyte (HTL) epitopes with interferon-inducing potential, and 7 linear B-cell epitopes. Furthermore, each of these epitopes was rigorously evaluated and confirmed to be antigenic, non-allergenic, and non-toxic. Collectively, the selected epitopes demonstrated broad predicted coverage across bovine leukocyte antigen (BoLA) alleles, thereby ensuring the potential for a wide-ranging immune response. To improve stability, folding, and immunogenicity, these epitopes were combined into a 590-amino-acid construct with the addition of the β-defensin-3 adjuvant and appropriate linkers. Moreover, the resultant construct's stability, solubility, and robust antigenic potential were validated by physicochemical profiling, confirming it's suitability as a promising vaccine candidate. Lastly, molecular docking and simulation analyses demonstrated strong binding to bovine TLR4, confirming the construct’s structural stability and compactness. Codon optimization and in silico cloning into the pET28a(+) vector indicated feasibility for high-yield expression and efficient purification in E. coli . These findings present the first conserved-region-based LSDV MEV construct tailored for Indian and regional viral strains. This design offers broad immune coverage, favourable biophysical properties, and strong receptor engagement, indicating its potential as an effective vaccine candidate. Future in vitro and in vivo validation will be critical to confirm immunogenicity, safety, and protective efficacy, with potential translational application for large-scale livestock immunization programs across LSDV-endemic regions.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.