Department of Biotechnology, School of Basic & Applied Sciences, Dayananda Sagar University, Bangalore, Karnataka, India.
Department of Biotechnology, Dayananda Sagar College of Engineering (Affiliated to Visvesvaraya Technological University, Belagavi), Bangalore, Karnataka, India.
J Biomol Struct Dyn. 2024;42(21):11792-11811. doi: 10.1080/07391102.2023.2265471. Epub 2023 Oct 5.
Infections caused by drug resistant bacteria is a silent detrimental pandemic affecting the global health care profoundly. Methicillin resistant (MRSA) is a pathogen that causes serious infections in different settings (community, hospital & veterinary) whose treatment remains highly challenging due to its powerful characteristics (antibiotic resistance strategies, virulence factors). In this study, we used reverse vaccinology (RV) approach and designed an immunogenic multi epitope vaccine (CV3Ag-antiMRSA) targeting three potential antigen candidates viz., encoding transpeptidase (PBP2a) protein responsible for conferring methicillin resistance and two virulence determinants - encoding gamma-hemolysin component A (a pore forming toxin) and encoding iron regulated surface determinant B (heme transport component that allows to scavenge iron from host hemoglobin and myoglobin). We employed an array of immunoinformatic tools/server to identify and use immunogenic epitopes (B cell and MHC class) to develop the chimeric subunit vaccine V4 (CV3Ag-antiMRSA) with immune modulating adjuvant and linkers. Based on different parameters, the vaccine construct V4 (CV3Ag-antiMRSA) was determined to be suitable vaccine (antigenic and non-allergen). Molecular docking and simulation of CV3Ag-antiMRSA with Toll Like Receptor (TLR2) predicted its immuno-stimulating potential. Finally, cloning of CV3Ag-antiMRSA construct into pet28a and pet30 vector displayed its feasibility for the heterologous expression in the expression system. This vaccine candidate (CV3Ag-antiMRSA) designed based on the MRSA genomes obtained from both animal and human hosts can be experimentally validated and thereby contribute to vaccine development to impart protection to both animal and human health.Communicated by Ramaswamy H. Sarma.
耐抗生素细菌感染是一种无声的、有害的全球健康危机。耐甲氧西林金黄色葡萄球菌（MRSA）是一种病原体，可在不同环境（社区、医院和兽医）中引起严重感染，由于其强大的特性（抗生素耐药策略、毒力因子），其治疗仍然极具挑战性。在这项研究中，我们使用反向疫苗学（RV）方法设计了一种针对三种潜在抗原候选物的免疫原性多表位疫苗（CV3Ag-antiMRSA），分别编码转肽酶（PBP2a）蛋白，该蛋白负责赋予耐甲氧西林的能力，以及两种毒力决定因素-编码γ-溶血素成分 A（一种形成孔的毒素）和编码铁调节表面决定簇 B（铁转运成分，使能够从宿主血红蛋白和肌红蛋白中掠夺铁）。我们利用一系列免疫信息学工具/服务器来识别和使用免疫原性表位（B 细胞和 MHC 类）来开发带有免疫调节佐剂和接头的嵌合亚单位疫苗 V4（CV3Ag-antiMRSA）。根据不同的参数，疫苗构建体 V4（CV3Ag-antiMRSA）被确定为合适的疫苗（抗原性和非变应原性）。CV3Ag-antiMRSA 与 Toll 样受体（TLR2）的分子对接和模拟预测了其免疫刺激潜力。最后，将 CV3Ag-antiMRSA 构建体克隆到 pet28a 和 pet30 载体中，显示了其在异源表达系统中的可行性。该疫苗候选物（CV3Ag-antiMRSA）是基于从动物和人类宿主中获得的 MRSA 基因组设计的，可以通过实验验证，从而有助于疫苗的开发，为动物和人类健康提供保护。
