Heidarinia Hana, Tajbakhsh Elahe, Rostamian Mosayeb, Momtaz Hassan
Department of Microbiology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.
Infectious Diseases Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Heliyon. 2023 Jul 25;9(8):e18614. doi: 10.1016/j.heliyon.2023.e18614. eCollection 2023 Aug.
Outer membrane protein W (OmpW) is a less-known antigen with potential immunogenic properties. The epitopes of this protein are not well-identified yet. Therefore, in the present study, B- and T-cell epitopes of OmpW were found using comprehensive in silico and partially in vitro studies. The T-cell (both class-I and class-II) and B-cell (both linear and conformational) epitopes were predicted and screened through many bioinformatics approaches including the prediction of IFN-γ production, immunogenicity, toxicity, allergenicity, human similarity, and clustering. A single 15-mer epitopic peptide containing a linear B-cell and both classes of T-cell epitopes were found and used for further assays. For in vitro assays, patient- and healthy control-derived peripheral blood mononuclear cells were stimulated with the 15-mer peptide, Phytohemagglutinin, or medium alone, and cell proliferation and IFN-γ production assays were performed. The bioinformatics studies led to mapping OmpW epitopes and introducing a 15-mer peptide. In vitro assays to some extent showed its potency in cell proliferation but not in IFN-γ induction, although the responses were not very expressive and faced some questions/limitations. In general, in the current study, we mapped the most immunogenic epitopes of OmpW that may be used for future studies and also assayed one of these epitopes in vitro, which was shown to have an immunogenicity potential. However, the induced immune responses were not strong which suggests that the present peptide needs a series of biotechnological manipulations to be used as a potential vaccine candidate. More studies in this field are recommended.
外膜蛋白W(OmpW)是一种鲜为人知但具有潜在免疫原性的抗原。该蛋白的表位尚未得到很好的鉴定。因此,在本研究中,通过全面的计算机模拟研究和部分体外研究发现了OmpW的B细胞和T细胞表位。通过多种生物信息学方法预测和筛选了T细胞(I类和II类)和B细胞(线性和构象)表位,包括预测干扰素-γ产生、免疫原性、毒性、致敏性、人源相似性和聚类分析。发现了一个包含线性B细胞表位和两类T细胞表位的15聚体表位肽,并用于进一步检测。对于体外检测,用该15聚体肽、植物血凝素或单独的培养基刺激患者和健康对照来源的外周血单个核细胞,并进行细胞增殖和干扰素-γ产生检测。生物信息学研究确定了OmpW表位并引入了一个15聚体肽。体外检测在一定程度上显示了其在细胞增殖方面的效力,但在干扰素-γ诱导方面未显示效力,尽管反应不太明显且存在一些问题/局限性。总体而言,在本研究中,我们确定了OmpW最具免疫原性的表位,这些表位可用于未来的研究,并且我们还在体外检测了其中一个表位,结果显示其具有免疫原性潜力。然而,诱导的免疫反应并不强烈,这表明目前的肽需要一系列生物技术操作才能用作潜在的疫苗候选物。建议在该领域进行更多研究。