Libentech Co., Ltd., Daejeon, Republic of Korea.
Department of Veterinary Medicine, Chungnam National University, Daejeon, Republic of Korea.
J Virol Methods. 2022 Jan;299:114347. doi: 10.1016/j.jviromet.2021.114347. Epub 2021 Oct 30.
The Middle East respiratory syndrome coronavirus (MERS-CoV), belonging to the family Coronaviridae and genus Betacoronavirus, has been recognized as a highly pathogenic virus. Due to the lack of therapeutic or preventive agents against MERS-CoV, developing an effective vaccine is essential for preventing a viral outbreak. To address this, we developed a recombinant S1 subunit of MERS-CoV spike protein fused with the human IgG4 Fc fragment (LV-MS1-Fc) in Chinese hamster ovary (CHO) cells. Thereafter, we identified the baculovirus gp64 signal peptide-directed secretion of LV-MS1-Fc protein in the extracellular fluid. To demonstrate the immunogenicity of the recombinant LV-MS1-Fc proteins, BALB/c mice were inoculated with 2.5 μg of LV-MS1-Fc. The inoculated mice demonstrated a significant humoral immune response, measured via total IgG and neutralizing antibodies. In addition, human dipeptidyl peptidase-4 (DPP4) transgenic mice vaccinated with LV-MS1-Fc showed the protective capacity of LV-MS1-Fc against MERS-CoV with no inflammatory cell infiltration. These data showed that the S1 and Fc fusion protein induced potent humoral immunity and antigen-specific neutralizing antibodies in mice, and conferred protection against coronavirus viral challenge, indicating that LV-MS1-Fc is an effective vaccine candidate against MERS-CoV infection.
中东呼吸综合征冠状病毒(MERS-CoV)属于冠状病毒科和β冠状病毒属,已被认为是一种高致病性病毒。由于缺乏针对 MERS-CoV 的治疗或预防药物,因此开发有效的疫苗对于预防病毒爆发至关重要。为了解决这个问题,我们在中华仓鼠卵巢(CHO)细胞中开发了一种与人 IgG4 Fc 片段融合的 MERS-CoV 刺突蛋白的重组 S1 亚单位(LV-MS1-Fc)。此后,我们鉴定了杆状病毒 gp64 信号肽指导的 LV-MS1-Fc 蛋白在细胞外液中的分泌。为了证明重组 LV-MS1-Fc 蛋白的免疫原性,用 2.5 μg 的 LV-MS1-Fc 接种 BALB/c 小鼠。接种的小鼠表现出显著的体液免疫反应,通过总 IgG 和中和抗体来衡量。此外,用人二肽基肽酶-4(DPP4)转基因小鼠接种 LV-MS1-Fc 后,LV-MS1-Fc 对 MERS-CoV 表现出保护能力,没有炎症细胞浸润。这些数据表明,S1 和 Fc 融合蛋白在小鼠中诱导了强烈的体液免疫和抗原特异性中和抗体,并赋予了针对冠状病毒病毒挑战的保护作用,表明 LV-MS1-Fc 是一种针对 MERS-CoV 感染的有效疫苗候选物。