Search for potential target site of nucleocapsid gene for the design of an epitope-based SARS DNA vaccine
Search for potential target site of nucleocapsid gene for the design of an epitope-based SARS DNA vaccine
复制标题
DOI：
10.1016/j.imlet.2008.03.003
复制
发表时间：
2008-06-15
影响因子：
4.4
通讯作者：
Park, Jae-Hak
中科院分区：
文献类型：
--
作者：
Dutta, Noton Kumar;Mazumdar, Kaushiki;Park, Jae-Hak
目前认为，严重急性呼吸综合征（SARS）冠状病毒的核衣壳蛋白（N）是最有前途的疫苗设计的候选抗原之一。本研究将SARS冠状病毒N蛋白的3个片段[NI（残基：1-422）; N2（残基：1-109）; N3（残基：110-422）]在大肠杆菌中表达，并用SARS患者恢复期血清进行分析。将SARS-N的三个基因片段[N1（1-1269 nt），N2（1-327 nt）和N3（328-1269 nt）]分别克隆到pVAX-1中，并通过电穿孔免疫BALB/c小鼠。用重组N1和N3特异性抗原检测体液免疫（酶联免疫吸附试验（ELISA））和细胞免疫（细胞增殖和CD 4+：CD 8+）。结果表明，在大肠杆菌中表达的N蛋白NI和N3片段，大肠杆菌能与SARS病人血清发生反应，而N2不能。SARS-CoV N1和N3 DNA疫苗能显著诱导小鼠产生特异性体液免疫和细胞免疫。此外，与N1疫苗相比，N3中的抗体应答（IgG和IgG 1，但不是IgG 2a）和细胞增殖的免疫应答水平显著更高，但在CD 4（+）：CD 8（+）试验中则不是。N蛋白抗原片段的鉴定为SARS冠状病毒DNA疫苗的设计提供了基础信息。本研究结果不仅表明pVax-N3 DNA免疫可作为诱导BALB/c小鼠产生抗体的潜在DNA疫苗途径，而且还表明这些SARS DNA疫苗的基因免疫可产生不同的免疫应答。(c)2008 Elsevier B. V.保留所有权利。
It is believed today that nucleocapsid protein (N) of severe acute respiratory syndrome (SARS)-CoV is one of the most promising antigen candidates for vaccine design. In this study, three fragments [NI (residues: 1-422); N2 (residues: 1-109); N3 (residues: 110-422)] of N protein of SARS-CoV were expressed in Escherichia coli and analyzed by pooled sera of convalescence phase of SARS patients. Three gene fragments [N1 (1-1269 nt), N2 (1-327 nt) and N3 (328-1269 nt)-expressing the same proteins of NI, N2 and N3, respectively] of SARS-N were cloned into pVAX-1 and used to immunize BALB/c mice by electroporation. Humoral (by enzyme-linked immunosorbent assay, ELISA) and cellular (by cell proliferation and CD4(+):CD8(+) assay) immunity was detected by using recombinant N1 and N3 specific antigen. Results showed that NI and N3 fragments of N protein expressed by E. coli were able to react with sera of SARS patients but N2 could not. Specific humoral and cellular immunity in mice could be induced significantly by inoculating SARS-CoV N1 and N3 DNA vaccine. In addition, the immune response levels in N3 were significantly higher for antibody responses (IgG and IgG1 but not IgG2a) and cell proliferation but not in CD4(+):CD8(+) assay compared to N1 vaccine. The identification of antigenic N protein fragments has implications to provide basic information for the design of DNA vaccine against SARS-CoV. The present results not only suggest that DNA immunization with pVax-N3 could be used as potential DNA vaccination approaches to induce antibody in BALB/c mice, but also illustrates that gene immunization with these SARS DNA vaccines can generate different immune responses. (c) 2008 Elsevier B.V. All rights reserved.