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[严重急性呼吸综合征相关冠状病毒刺突蛋白S1结构域在小鼠体内引发的强效中和抗体]

[Potent neutralization antibody elicited in mice by SARS-associated coronavirus spike protein S1 domain].

作者信息

Zhang Yun, Yang Fan, Li Yan-han, Li Wen-hui, Tu Xin-ming, Wei Qiang, Zhu Hua, Liu Li, Wang Heng, Qin Chuan, Yuan Guo-yong, He Wei, Wang Shu-hui

机构信息

Department of Etiology, Institute of Basic Medical Science, CAMS and PUMC, Beijing 100005, China.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2004 Sep;18(3):258-60.

Abstract

OBJECTIVE

To study the antigenicity of SARS associated coronavirus (CoV) spike S1 (12-672Aa) domain.

METHODS

BALB/c mice were immunized with a plasmid bearing codon-optimized SARS-CoV (Tor2 strain) S1 domain and then boosted with purified S1 protein; the SARS-CoV specific IgG antibody was tested by ELISA and neutralization antibody was determined by in vitro microneutralization assay.

RESULTS

S1 domain of SARS-CoV spike, which has been demonstrated harboring the receptor binding domain, successfully elicited SARS-CoV specific IgG antibody in mouse after combined immunization with DNA and purified S1 protein; the antibody elicited solely by S1 could potently neutralize SARS-CoV (HKU-39849) in vitro, 50% of 1 000 TCID50 SARS-CoV challenged cells were protected from viral infection by a 1:1499.68 dilution of mice sera immunized with S1 protein, but negative control sera showed no protection.

CONCLUSION

S1 domain of SARS-CoV spike protein, which is responsible for receptor binding, can efficiently and sufficiently induce highly potent neutralizing antibody in mice. This result suggested that S1 domain could be an effective subunit vaccines against SARS-CoV.

摘要

目的

研究严重急性呼吸综合征相关冠状病毒(CoV)刺突S1(12 - 672Aa)结构域的抗原性。

方法

用携带密码子优化的严重急性呼吸综合征冠状病毒(Tor2株)S1结构域的质粒免疫BALB/c小鼠,然后用纯化的S1蛋白进行加强免疫;通过酶联免疫吸附测定(ELISA)检测严重急性呼吸综合征冠状病毒特异性IgG抗体,并用体外微量中和试验测定中和抗体。

结果

严重急性呼吸综合征冠状病毒刺突的S1结构域已被证明含有受体结合结构域,在与DNA和纯化的S1蛋白联合免疫后,成功在小鼠体内诱导出严重急性呼吸综合征冠状病毒特异性IgG抗体;仅由S1诱导产生的抗体在体外能有效中和严重急性呼吸综合征冠状病毒(HKU - 39849),用S1蛋白免疫的小鼠血清以1:1499.68的稀释度可保护50%受1000半数组织培养感染剂量(TCID50)严重急性呼吸综合征冠状病毒攻击的细胞免受病毒感染,但阴性对照血清无保护作用。

结论

严重急性呼吸综合征冠状病毒刺突蛋白的S1结构域负责受体结合,能在小鼠体内有效且充分地诱导出高效中和抗体。该结果表明S1结构域可能是一种有效的抗严重急性呼吸综合征冠状病毒亚单位疫苗。

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