利用保守的E2蛋白和NS3 T淋巴细胞表位设计针对经典猪瘟病毒的mRNA疫苗的免疫信息学研究

Immunoinformatics design of an mRNA vaccine against classical swine fever virus using conserved E2 protein and NS3 T-lymphocyte epitopes.

作者信息

Banico Edward C, Sira Ella Mae Joy S, Fajardo Lauren Emily, Orosco Fredmoore L

机构信息

irology and Vaccine Research Program, Industrial Technology Development Institute, Department of Science and Technology, Taguig City, 1634, Philippines.

S&T Fellows Program, Department of Science and Technology, Taguig City, 1634, Philippines.

出版信息

Bioimpacts. 2025 Apr 6;15:30778. doi: 10.34172/bi.30778. eCollection 2025.

DOI:10.34172/bi.30778

PMID:40584901

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12204779/

Abstract

INTRODUCTION

The classical swine fever virus (CSFV) causes significant economic losses in the livestock industry. While the existing E2 marker vaccine offers protection against infections, it is characterized by delayed immunity and reduced effectiveness over time. Optimizing the existing vaccine is crucial to better control CSFV outbreaks worldwide. This study aimed to improve the existing E2 marker vaccine for CSFV by integrating NS3 T lymphocyte-inducing epitopes into the conserved E2 protein sequence and using mRNA technology for vaccine delivery.

METHODS

The design and evaluation of the vaccine were carried out exclusively through methods. T lymphocyte epitopes were identified from the CSFV NS3 protein using multiple epitope prediction tools. A vaccine construct was formed after linking the predicted NS3 epitopes, E2 protein, and an immunogenic adjuvant. Molecular docking and dynamics simulations were performed to analyze the interaction between the adjuvant used and its immune receptor. Signal peptides were incorporated into the design, and mRNA sequences with varying codon usage biases were generated using LinearDesign. The mRNA sequence with minimum free energy (MFE) and codon adaptation index (CAI) closest to the controls was selected as the final design.

RESULTS

Twenty epitopes with high binding affinity to major histocompatibility complexes (MHCs) were identified from the CSFV NS3 protein. The vaccine construct with swine CD154 adjuvant demonstrated high antigenicity, making it the optimal choice for the final vaccine design. Molecular docking and dynamics simulations confirmed the adjuvant's strong affinity and stable interaction with its canonical receptor, swine CD40. Moreover, the final vaccine design exhibited higher populations of lymphocytes and antibodies compared to the components of the commercialized E2 marker vaccine in immune simulation. The final mRNA vaccine sequence exhibited a higher MFE and CAI than the two licensed mRNA vaccine controls.

CONCLUSION

The mRNA vaccine designed in this study serves as a potential CSFV vaccine candidate. and validation is needed to confirm its efficacy.

摘要

引言

经典猪瘟病毒（CSFV）给畜牧业造成了巨大经济损失。虽然现有的E2标记疫苗能提供抗感染保护，但其特点是免疫反应延迟且随着时间推移效力降低。优化现有疫苗对于在全球范围内更好地控制CSFV疫情至关重要。本研究旨在通过将NS3 T淋巴细胞诱导表位整合到保守的E2蛋白序列中，并使用mRNA技术进行疫苗递送，来改进现有的CSFV E2标记疫苗。

方法

疫苗的设计和评估完全通过[具体方法未给出]方法进行。使用多种表位预测工具从CSFV NS3蛋白中鉴定T淋巴细胞表位。将预测的NS3表位、E2蛋白和一种免疫佐剂连接后形成疫苗构建体。进行分子对接和动力学模拟以分析所用佐剂与其免疫受体之间的相互作用。在设计中加入信号肽，并使用LinearDesign生成具有不同密码子使用偏好的mRNA序列。选择具有最低自由能（MFE）且密码子适应指数（CAI）最接近对照的mRNA序列作为最终设计。

结果

从CSFV NS3蛋白中鉴定出20个与主要组织相容性复合体（MHC）具有高结合亲和力的表位。含有猪CD154佐剂的疫苗构建体表现出高抗原性，使其成为最终疫苗设计的最佳选择。分子对接和动力学模拟证实了该佐剂与其经典受体猪CD40具有强亲和力和稳定相互作用。此外，在免疫模拟中，最终疫苗设计与商业化E2标记疫苗的成分相比，表现出更高的淋巴细胞和抗体数量。最终的mRNA疫苗序列比两种已获许可的mRNA疫苗对照具有更高的MFE和CAI。