Antigenicity and immunogenicity of SARS-CoV S protein receptor-binding domain stably expressed in CHO cells.

Antigenicity and immunogenicity of SARS-CoV S protein receptor-binding domain stably expressed in CHO cells.
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DOI:
10.1016/j.bbrc.2009.05.003
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发表时间:
2009-07-10
影响因子:
3.1
通讯作者:
Jiang, Shibo
Jiang, Shibo
中科院分区:
生物学4区
文献类型:
--
作者:
Du, Lanying;Zhao, Guangyu;Li, Lin;He, Yuxian;Zhou, Yusen;Zheng, Bo-Jian;Jiang, Shibo

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SARS冠状病毒S蛋白受体结合域(receptor-binding domain,RBD)含有多个构象依赖性表位,可诱导中和抗体应答。在此,我们使用CHO-K1细胞来建立用于稳定表达193-mer(残基318-510)RBD(RBD 193-CHO)的细胞系,并测定其抗原性和免疫原性。我们发现RBD 193-CHO与一组识别RBD中各种构象和线性表位的六种单克隆抗体强烈反应,表明该重组蛋白保持了完整的构象和良好的抗原性。用RBD 193-CHO免疫小鼠导致诱导高滴度的RBD特异性中和抗体和有效的表达IL-4的T细胞应答。RBD 193-CHO诱导的免疫保护了大多数接种疫苗的小鼠免受SARS-CoV攻击。这些结果表明,在建立的稳定的细胞系中产生的重组RBD保持了强的免疫原性,具有作为有效和经济的亚单位SARS疫苗的高潜力。
The receptor-binding domain (RBD) of SARS coronavirus (SARS-CoV) spike (S) protein contains multiple conformation-dependent epitopes that induce neutralizing antibody responses. Here we used CHO-K1 cells to establish a cell line for stable expression of a 193-mer (residues 318–510) RBD (RBD193-CHO) and determined its antigenicity and immunogenicity. We found that RBD193-CHO reacted strongly with a panel of six monoclonal antibodies recognizing various conformational and linear epitopes in RBD, suggesting that this recombinant protein maintains intact conformation and good antigenicity. Immunization of mice with RBD193-CHO resulted in induction of high titers of RBD-specific neutralizing antibodies and potent IL-4-expressing T cell responses. RBD193-CHO induced immunity that protected a majority of the vaccinated mice from SARS-CoV challenge. These results suggest that the recombinant RBD produced in an established stable cell line maintains strong immunogenicity with high potential for use as an effective and economic subunit SARS vaccine.
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