Faculty of Biotechnology and Genetic Engineering, Sylhet Agricultural University, Bangladesh.
Faculty of Biotechnology and Genetic Engineering, Sylhet Agricultural University, Bangladesh; Department of Animal and Fish Biotechnology, Sylhet Agricultural University, Bangladesh.
Mol Biochem Parasitol. 2024 Dec;260:111655. doi: 10.1016/j.molbiopara.2024.111655. Epub 2024 Nov 7.
The parasite Toxoplasma gondii, or T. gondii, is zoonotic that both individuals as well as animals can contract resulting in toxoplasmosis, a life-threatening illness. We used an immunoinformatic technique in our research to construct a vaccine with multi-epitopes so that it can decrease the devastating impact caused by this dangerous parasite. In order to construct the vaccine, GRA6 and MIC3 proteins were targeted, which are engaged in T. gondii identification, infection, and immune response. Novel epitopes for linear B lymphocytes (LBL), cytotoxic T lymphocytes (CTL), and helper T lymphocytes (HTL) were found by epitope mapping, every anticipated epitope was assessed through rigorous screening to determine the top choices for epitopes which were entirely preserved, very antigenic in nature, nonallergenic, and nontoxic. 4 CTLs, 3 HTLs and 4 LBL epitopes were chosen and combined along with proper linkers and adjuvants to design a vaccine with several epitopes. Linkers as well as adjuvants were provided to make the vaccine more immunogenic, antigenic, and stable. The proposed vaccination was identified to possess the necessary biophysical properties, be soluble, extremely antigenic, and non-allergic. Reliability of the vaccine design was demonstrated by secondary along with tertiary structure prediction. It was anticipated that the vaccine's three-dimensional structure would likely link up with TLR-2 and TLR-4 via the investigation of molecular docking. TLR-2 and TLR-4 are crucial for the parasite's invasion and the body's response. In our docking investigation, both TLRs demonstrated strong binding affinities utilizing the vaccine structure. After that, the vaccine construct's elevated expression rate, which was observed in Escherichia coli strain K12, was confirmed by an investigation using in silico cloning and codon adaptation. The results of the research are really encouraging and some properties of the vaccine were found to be significantly better than existing the T. gondii multi-epitope vaccination based on the same proteins. Nonetheless, in vivo trials are strongly suggested for potential future studies.
寄生虫刚地弓形虫，或弓形虫，是一种人畜共患的寄生虫，可以感染个体和动物，导致弓形体病，这是一种危及生命的疾病。我们在研究中使用免疫信息学技术构建了一种带有多种表位的疫苗，以减少这种危险寄生虫造成的破坏性影响。为了构建疫苗，我们选择了 GRA6 和 MIC3 蛋白，它们参与了弓形虫的识别、感染和免疫反应。通过表位作图发现了新型线性 B 淋巴细胞（LBL）、细胞毒性 T 淋巴细胞（CTL）和辅助 T 淋巴细胞（HTL）的表位，通过严格筛选评估了每个预期表位，以确定完全保留、高度抗原性、无变应原性和非毒性的表位的首选。选择了 4 个 CTL、3 个 HTL 和 4 个 LBL 表位，并与适当的接头和佐剂结合，设计了一种带有多个表位的疫苗。接头和佐剂的提供使疫苗更具免疫原性、抗原性和稳定性。所提出的疫苗被认为具有必要的生物物理特性，可溶、高度抗原性和无变应原性。通过二级和三级结构预测证明了疫苗设计的可靠性。通过分子对接研究，预计疫苗的三维结构可能通过与 TLR-2 和 TLR-4 结合。TLR-2 和 TLR-4 对寄生虫的入侵和机体的反应至关重要。在我们的对接研究中，两种 TLR 都利用疫苗结构表现出很强的结合亲和力。之后，通过在大肠杆菌 K12 菌株中进行的体外克隆和密码子适应度分析，证实了疫苗构建体的高表达率。研究结果令人鼓舞，发现一些疫苗的特性明显优于基于相同蛋白的现有的弓形虫多表位疫苗。然而，强烈建议进行体内试验，以进行未来的潜在研究。
