School of Biotechnology, KIIT Deemed to be University, Bhubaneswar, Odisha, India; National Veterinary Institute, Debre Zeit, Ethiopia.
School of Biotechnology, KIIT Deemed to be University, Bhubaneswar, Odisha, India.
Acta Trop. 2021 May;217:105870. doi: 10.1016/j.actatropica.2021.105870. Epub 2021 Feb 24.
Tuberculosis (TB) is a complex infectious bacterial disease, which has evolved with highly successful mechanisms to interfere with host defenses and existing classes of antibiotics to resist eradication. The single obtainable TB vaccine, Bacille Calmette-Guerin (BCG) has failed to provide regular defense for respiratory TB in adults. In this study, a bioinformatics and immunoinformatics approach was applied on Mycobacterium tuberculosis (Mtb) H37Rv proteomes to discover the potential subunit vaccine candidates that elicit both tuberculosis-specific T-cells and B-cell immune response. A total of 4049 proteins of MtbH37RvMtbH37Rv were retrieved and subjected to in silico sequence-based analysis. Finally, five (P9WL69 (Rv2599), P9WIG1 (Rv0747), P9WLQ1 (Rv1987), O53608 (Rv0063), O06624 (Rv1566c)) novel putative proteins were selected. Among the five putative antigenic vaccine candidates, P9WL69 protein was selected for the ex-vivo validation study. The P9WL69 protein encoding gene was amplified and cloned on pET21b vector. The success of the recombinant clone (pET21b-RV2599) was confirmed by colony PCR, insert release test and sequencing. Furthermore, the identified epitopes of the P9WL69 protein were considered for in silico docking and molecular dynamics simulation study using Toll-like Receptors (TLRs) (TLR-2, TLR-4, TLR-9), Mannose receptor, and Myeloid differentiation 88 (MYD88) to understand their binding affinity towards the development of immunogenic vaccines against tuberculosis.
结核病（TB）是一种复杂的细菌性传染病，它已经进化出了高度成功的机制来干扰宿主防御和现有的抗生素类别，以抵抗根除。唯一可用的结核病疫苗卡介苗（BCG）未能为成人呼吸道结核病提供常规防御。在这项研究中，我们应用生物信息学和免疫信息学方法对结核分枝杆菌（Mtb）H37Rv 蛋白质组进行分析，以发现能够引发结核特异性 T 细胞和 B 细胞免疫反应的潜在亚单位疫苗候选物。共检索到 4049 种 MtbH37Rv 蛋白质，并对其进行了基于序列的计算机分析。最后，选择了五个（P9WL69（Rv2599）、P9WIG1（Rv0747）、P9WLQ1（Rv1987）、O53608（Rv0063）、O06624（Rv1566c））新型潜在蛋白作为候选抗原。在这五个潜在的抗原性疫苗候选物中，选择 P9WL69 蛋白进行体外验证研究。扩增并克隆 P9WL69 蛋白编码基因至 pET21b 载体上。通过菌落 PCR、插入片段释放试验和测序验证了重组克隆（pET21b-RV2599）的成功构建。此外，还对 P9WL69 蛋白的鉴定表位进行了计算机对接和分子动力学模拟研究，使用 Toll 样受体（TLR）（TLR-2、TLR-4、TLR-9）、甘露糖受体和髓样分化 88（MYD88）来了解它们与结核免疫疫苗开发的结合亲和力。
