Hasani Sharareh Mohammad, Behdani Mahdi, Amirkhani Zohreh, Rahimmanesh Ilnaz, Esmaeilifallah Mahsa, Zaker Erfan, Nikpour Parvaneh, Fadaie Mahmood, Ghafouri Elham, Naderi Shamsi, Khanahmad Hossein
Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Department of Biotechnology, Biotechnology Research Center, Venom and Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, Iran.
Res Pharm Sci. 2024 Oct 22;19(5):573-590. doi: 10.4103/RPS.RPS_82_24. eCollection 2024 Oct.
The global emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has prompted widespread concern. Bacteriophages have recently gained attention as a cost-effective and stable alternative for vaccine development due to their adjuvant properties. This study aimed to design and validate a poly epitope composed of viral proteins.
SARS-CoV-2 proteins (spike, nucleocapsid, membrane, envelope, papain-like protease, and RNA-dependent RNA polymerase) were selected for analysis. Immunoinformatic methods were employed to predict B and T cell epitopes, assessing their antigenicity, allergenicity, and toxicity. Epitopes meeting criteria for high antigenicity, non-allergenicity, and non-toxicity were linked to form poly epitopes. These sequences were synthesized and cloned into pHEN4 plasmids to generate Poly1 and Poly2 phagemid vectors. Recombinant Poly1 and Poly2 phages were produced by transforming M13ΔIII plasmids and phagemid vectors into . Female Balb/c mice were immunized with a cocktail of Poly1 and Poly2 phages, and their serum was collected for ELISA testing. Interferon-gamma (IFN-γ) testing was performed on spleen-derived lymphocytes to evaluate immune system activation.
FINDINGS/RESULTS: Recombinant Poly1 and Poly2 phages were produced, and their titer was determined as 10 PFU/mL. Efficient humoral immune responses and cellular immunity activation in mice were achieved following phage administration.
Poly epitopes displayed on phages exhibit adjuvant properties, enhancing humoral and cellular immunity in mice. This suggests that phages could serve as adjuvants to bolster immunity against SARS-Cov-2. Recombinant phages could be applied as effective candidates for injectable and oral vaccine development strategies.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）在全球出现引发了广泛关注。由于其佐剂特性，噬菌体最近作为一种经济高效且稳定的疫苗开发替代方案受到关注。本研究旨在设计并验证一种由病毒蛋白组成的多表位。
选择SARS-CoV-2蛋白（刺突蛋白、核衣壳蛋白、膜蛋白、包膜蛋白、木瓜样蛋白酶和RNA依赖性RNA聚合酶）进行分析。采用免疫信息学方法预测B细胞和T细胞表位，评估其抗原性、致敏性和毒性。符合高抗原性、无致敏性和无毒性标准的表位被连接形成多表位。合成这些序列并克隆到pHEN4质粒中以产生Poly1和Poly2噬菌粒载体。通过将M13ΔIII质粒和噬菌粒载体转化到……中来产生重组Poly1和Poly2噬菌体。用Poly1和Poly2噬菌体混合物免疫雌性Balb/c小鼠，并收集其血清进行ELISA检测。对脾来源的淋巴细胞进行干扰素-γ（IFN-γ）检测以评估免疫系统激活情况。
产生了重组Poly1和Poly2噬菌体，其滴度测定为10 PFU/mL。噬菌体给药后在小鼠中实现了有效的体液免疫反应和细胞免疫激活。
噬菌体展示的多表位具有佐剂特性，可增强小鼠的体液免疫和细胞免疫。这表明噬菌体可作为佐剂增强针对SARS-CoV-2的免疫力。重组噬菌体可作为注射和口服疫苗开发策略的有效候选物。
