Abstract：
Clostridium perfringens is a bacterium that causes gastrointestinal diseases in humans and animals. The several powerful toxins such as alpha toxin (CPA), beta toxin (CPB), enterotoxin (CPE), Epsilon toxin (ETX), and theta toxin, play a major role in its pathogenesis. Traditional vaccine development methods are time-consuming and costly. In silico approaches offer an alternative strategy for designing vaccines by analyzing biological data and predicting immunogenic peptides. In this study, computational tools were utilized to design a RNA vaccine targeting C. perfringens toxins. Toxin protein sequences were retrieved and their linear B-cell, MHCI, and MHCII binding epitopes were predicted. Allergenicity, toxigenicity, and IFN-γ induction were assessed to select non-allergenic, non-toxic, and IFN-γ-inducing epitopes. Molecular docking was performed to identify epitopes that fit within the binding cleft of MHC alleles. A final peptide vaccine construct was designed with selected epitopes separated by a linker sequence. The antigenicity and physicochemical properties of the vaccine were evaluated. Immune response simulation showed enhanced secondary and tertiary immune responses, increased levels of immunoglobulins, cytotoxic T lymphocytes, helper T lymphocytes, macrophage activity, and elevated levels IFN-γ and interleukin-2. Docking analysis was done to assess interactions between the vaccine structure and Toll-like receptors. Codon optimization was performed, and a final RNA vaccine construct was designed. The secondary structure of the RNA vaccine was predicted and validated. Overall, this study demonstrates the potential of in silico approaches for designing an RNA vaccine against C. perfringens toxins, contributing to improved prevention and control of associated diseases.
摘要：
产气荚膜梭菌是一种引起人和动物胃肠道疾病的细菌。几种强大的毒素，如α毒素(CPA)，β毒素（CPB），肠毒素（CPE），ε毒素（ETX），和theta毒素,在其发病机制中起主要作用。传统的疫苗开发方法耗时且昂贵。计算机模拟方法通过分析生物学数据和预测免疫原性肽为设计疫苗提供了替代策略。在这项研究中,利用计算工具设计靶向产气荚膜梭菌毒素的RNA疫苗。检索毒素蛋白序列及其线性B细胞，预测MHCI和MHCII结合表位。致敏性,毒性，和IFN-γ诱导进行评估，以选择非过敏性，无毒,和IFN-γ诱导表位。进行分子对接以鉴定适合于MHC等位基因的结合间隙内的表位。设计最终的肽疫苗构建体,其中选择的表位由接头序列分开。评价了疫苗的抗原性和理化性质。免疫反应模拟显示增强的二级和三级免疫反应，免疫球蛋白水平升高，细胞毒性T淋巴细胞，辅助性T淋巴细胞，巨噬细胞活性,IFN-γ和白介素-2水平升高。进行对接分析以评估疫苗结构与Toll样受体之间的相互作用。进行了密码子优化，并设计了最终的RNA疫苗构建体。预测并验证了RNA疫苗的二级结构。总的来说,这项研究证明了设计抗产气荚膜梭菌毒素的RNA疫苗的方法的潜力，有助于改善相关疾病的预防和控制。
