Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.
Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.
Microb Pathog. 2022 Sep;170:105687. doi: 10.1016/j.micpath.2022.105687. Epub 2022 Jul 31.
and Introduction: SARS-CoV-2 is currently considered as the most challenging issue in the field of health and medicine by causing a global pandemic. Vaccines are counted as a promising candidate to terminate this deadly pandemic. Various structural proteins in SARS-CoV-2 have recently drawn attention to be utilized as candidate vaccines to stimulate immune responses against COVID-19.
In current study, the RBD protein was cloned and expressed in E. coli host. Then, the expressed RBD protein was purified and its characterizations were evaluated through various methods. Gold nanoparticles, which were utilized as a carrier for candidate Nano-vaccine, were synthesized via oxidation-reduction reaction. While Gold NPs-conjugated RBD was injected into the second treatment group, in the first candidate vaccine, RBD was injected into the first treatment group solely. Complete and Incomplete Freud's Adjuvant were also utilized for both treatment groups to enhance the immune responses against RBD antigen. Immunizations were repeated 2 times in 14-day intervals to boost the immune system of BALB/c mice. The humoral and cell-mediated immune responses were examined through immune and cytokine assays.
Our outcomes demonstrate that strong short-term humoral immunity (IgM) was induced in both the first and second treatment group, while long-term humoral responses (IgG) were only observed in the second treatment group. While stronger short- and long-term humoral (IgM and IgG, respectively) were observed in the second treatment group, particular cytokines production (TNF-ɑ and IFN-γ) as a marker of cell-mediated responses were significantly higher in the first treatment group.
Our study results show the high potentiality of RBD protein as an appropriate stimulating antigen in vaccine synthesis and testifies RBD-based candidate vaccines to control the COVID-19 pandemic. Our outcomes also recommend that Nano-vaccines can be more suitable candidates when stronger long-term immune responses matter.
目前,SARS-CoV-2 被认为是健康和医学领域最具挑战性的问题,导致了全球大流行。疫苗被认为是终止这一致命大流行的有希望的候选者。SARS-CoV-2 的各种结构蛋白最近引起了人们的关注,可作为候选疫苗,以刺激针对 COVID-19 的免疫反应。
在本研究中,RBD 蛋白在大肠杆菌宿主中被克隆和表达。然后,表达的 RBD 蛋白被纯化,并通过各种方法评估其特性。金纳米粒子被用作候选纳米疫苗的载体,通过氧化还原反应合成。当金纳米粒子偶联的 RBD 被注射到第二治疗组时,在第一候选疫苗中,仅将 RBD 注射到第一治疗组中。完全和不完全弗氏佐剂也被用于两个治疗组,以增强针对 RBD 抗原的免疫反应。免疫接种在 14 天的间隔内重复两次,以增强 BALB/c 小鼠的免疫系统。通过免疫和细胞因子测定检查体液和细胞介导的免疫反应。
我们的结果表明,两个治疗组均诱导了强烈的短期体液免疫(IgM),而仅在第二治疗组观察到长期体液免疫(IgG)。虽然第二治疗组观察到更强的短期和长期体液免疫(分别为 IgM 和 IgG),但第一治疗组细胞介导反应的特定细胞因子(TNF-ɑ和 IFN-γ)的产生更高。
我们的研究结果表明,RBD 蛋白作为疫苗合成中的合适刺激抗原具有很高的潜力,并证明了基于 RBD 的候选疫苗可以控制 COVID-19 大流行。我们的结果还表明,当需要更强的长期免疫反应时,纳米疫苗可能是更合适的候选者。