Kumar Sachin, Moolchandani Shubham
Department of Pharmacology, Delhi Institute of Pharmaceutical Sciences and Research, Delhi Pharmaceutical Sciences and Research University, Pushp Vihar, MB Road, New Delhi, 110017 India.
Department of Pharmaceutical Biotechnology, School of Pharmaceutical Sciences, Delhi Pharmaceutical Sciences and Research University, Pushp Vihar, MB Road, New Delhi, 110017 India.
In Silico Pharmacol. 2025 Jun 9;13(2):83. doi: 10.1007/s40203-025-00365-x. eCollection 2025.
This study presents an in-silico approach to design a vaccine targeting Listeriolysin O (LLO) exotoxin in , a significant foodborne pathogen. Utilizing bioinformatics tools, we identified and prioritized B-cell and T-cell epitopes from the LLO sequence, ensuring non-toxicity, immunogenicity, non-allergenicity, and water solubility. The final vaccine construct, comprising 315 amino acids, was developed by combining selected epitopes with appropriate linkers and an adjuvant. Physicochemical characterization revealed favorable properties, including stability and solubility. Immune simulation using the C-ImmSim server predicted robust cellular and humoral responses, with significant increases in antibody levels and cytokine production within five days of administration in an in-silico study. Structural analysis of the vaccine construct yielded a refined 3D model with about 95% of residues in most favored regions of the Ramachandran plot. Protein-protein docking analysis using the ClusPro server predicted significant binding affinity between the vaccine construct and MHC-II receptors, with multiple hydrogen bonds and salt bridges contributing to stable interactions, as confirmed by PDBsum interaction analysis. Codon optimization for expression in resulted in a high Codon Adaptation Index (0.977) and suitable GC content (53.59%). The optimized sequence was successfully integrated into a pET28a (+) vector in-silico. While these computational results are promising, experimental validation is necessary to confirm the vaccine's immunogenicity, safety, and efficacy against infection. This study demonstrates the potential of in-silico approaches in accelerating vaccine development against challenging pathogens and provides a foundation for further research into listeriosis prevention.
本研究提出了一种计算机模拟方法，用于设计针对重要食源性病原体中李斯特菌溶血素O（LLO）外毒素的疫苗。利用生物信息学工具，我们从LLO序列中识别并确定了B细胞和T细胞表位的优先级，确保其无毒性、免疫原性、无致敏性和水溶性。通过将选定的表位与合适的连接子和佐剂相结合，开发出了最终由315个氨基酸组成的疫苗构建体。理化特性分析显示出良好的性质，包括稳定性和溶解性。在一项计算机模拟研究中，使用C-ImmSim服务器进行的免疫模拟预测了强烈的细胞和体液反应，给药后五天内抗体水平和细胞因子产生显著增加。对疫苗构建体的结构分析产生了一个精细的三维模型，在拉氏图的最有利区域中约95%的残基处于该区域。使用ClusPro服务器进行的蛋白质-蛋白质对接分析预测疫苗构建体与MHC-II受体之间具有显著的结合亲和力，多个氢键和盐桥有助于稳定相互作用，PDBsum相互作用分析证实了这一点。为在[具体物种]中表达而进行的密码子优化产生了较高的密码子适应指数（0.977）和合适的GC含量（53.59%）。优化后的序列在计算机模拟中成功整合到pET28a（+）载体中。虽然这些计算结果很有前景，但仍需要进行实验验证，以确认该疫苗对[具体病原体]感染的免疫原性、安全性和有效性。本研究证明了计算机模拟方法在加速针对具有挑战性的病原体的疫苗开发方面的潜力，并为进一步研究预防李斯特菌病提供了基础。
