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<p>Classical swine fever (CSF) is a highly contagious swine viral disease caused by classical swine fever virus (CSFV). Although modern CSFV vaccines are available, they tend to induce a delayed onset of immunity and show reduced efficacy over time. These limitations highlight the need to develop more effective vaccine alternatives to ensure better protection and management of the virus. This study aimed to design an epitope-based subunit vaccine targeting CSFV using reverse vaccinology. Immunogenic regions of the NS3 and E2 proteins, previously identified immunogenic proteins of the virus, were identified and integrated to form a vaccine construct. Specifically, five cytotoxic T lymphocyte (CTL) and five helper T lymphocyte (HTL) epitopes with the most stable binding to MHC molecules in molecular docking and dynamics were selected from all potential T lymphocyte-inducing epitopes within the NS3 sequence. Meanwhile, five linear B lymphocyte (LBL) epitopes were predicted with high confidence in the E2 protein by multiple LBL epitope prediction tools. Multiple linkers were used to connect epitopes forming a multi-epitope construct. Six different adjuvants were incorporated into the construct and subjected to multiple sequence- and structure-based evaluations. Among the designed vaccine, the one adjuvanted with phenol-soluble modulin α4 (pmα4) exhibited the highest B lymphocyte population and antibody titers in immune simulation, with satisfaction on the critical parameters including antigenicity, non-allergenicity, and solubility. Moreover, 23 conformational epitopes were identified on this construct following conformational epitopes screening using Ellipro. Stable interactions with Toll-like receptors (TLR2-TLR6) were also observed in molecular docking and dynamics. These findings highlight the pmα4-adjuvanted construct as the most promising vaccine candidate, demonstrating strong immunogenic potential, structural stability, and effective receptor binding—supporting its advancement to in vitro and in vivo validation for potential application in CSFV vaccine development.</p>

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Reverse vaccinology-aided design of an epitope-based subunit vaccine against classical swine fever virus

  • 期刊论文

  • 发表日期：

    2025年8月1日

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  • Edward C. Banico,
  • Ella Mae Joy S. Sira,
  • Lauren Emily Fajardo,
  • Nyzar Mabeth O. Odchimar,
  • Fredmoore L. Orosco

摘要

Classical swine fever (CSF) is a highly contagious swine viral disease caused by classical swine fever virus (CSFV). Although modern CSFV vaccines are available, they tend to induce a delayed onset of immunity and show reduced efficacy over time. These limitations highlight the need to develop more effective vaccine alternatives to ensure better protection and management of the virus. This study aimed to design an epitope-based subunit vaccine targeting CSFV using reverse vaccinology. Immunogenic regions of the NS3 and E2 proteins, previously identified immunogenic proteins of the virus, were identified and integrated to form a vaccine construct. Specifically, five cytotoxic T lymphocyte (CTL) and five helper T lymphocyte (HTL) epitopes with the most stable binding to MHC molecules in molecular docking and dynamics were selected from all potential T lymphocyte-inducing epitopes within the NS3 sequence. Meanwhile, five linear B lymphocyte (LBL) epitopes were predicted with high confidence in the E2 protein by multiple LBL epitope prediction tools. Multiple linkers were used to connect epitopes forming a multi-epitope construct. Six different adjuvants were incorporated into the construct and subjected to multiple sequence- and structure-based evaluations. Among the designed vaccine, the one adjuvanted with phenol-soluble modulin α4 (pmα4) exhibited the highest B lymphocyte population and antibody titers in immune simulation, with satisfaction on the critical parameters including antigenicity, non-allergenicity, and solubility. Moreover, 23 conformational epitopes were identified on this construct following conformational epitopes screening using Ellipro. Stable interactions with Toll-like receptors (TLR2-TLR6) were also observed in molecular docking and dynamics. These findings highlight the pmα4-adjuvanted construct as the most promising vaccine candidate, demonstrating strong immunogenic potential, structural stability, and effective receptor binding—supporting its advancement to in vitro and in vivo validation for potential application in CSFV vaccine development.