重组SARS-CoV S蛋白表位肽疫苗的研究

发布时间：2018-01-04 07:16

本文关键词：重组SARS-CoV S蛋白表位肽疫苗的研究　出处：《吉林大学》2005年博士论文　论文类型：学位论文

【摘要】：2002～2003 年爆发流行的传染性非典型肺炎(SARS)共造成29 个国家的8098 例感染,774 例死亡,引起了广泛关注。SAR.S 相关冠状病毒(SAR.S.CoV)是SARS 的病原体。棘突蛋白(S 蛋白)是SARS—CoV 的外壳成分,为SARS-CoV 识别和结合宿主细胞的关键结构,可以诱生SAR.S.CoV 的中和抗体,因此可作为抗SARS.CoV 药物及疫苗研究的靶点。为了研制预防SARS 疾病的疫苗,我们采用生物信息学方法对S 蛋白结构和特性做预测,选定了可能包含B 淋巴细胞表位区为168aa-216aa 肽段(S_2)和726aa.843aa 肽段(S_6)。在此基础上,构建出S_2 基因六聚体(PS_2基因)和分子伴侣10-s6融合基因(C86 基因),采用大肠杆菌高效表达了重组PS_2 和CS_6 蛋白,并用Ni2+亲合层析方法获得了纯化的重组蛋白。重组PS_2 蛋白在小鼠诱生的高滴度抗体以IgGl 为主,可识别SARS-CoV S 蛋白的S1 亚单位全长多肽(S1 多肽)和灭活的SARS—CoV。此外,重组PS_2蛋白能在体外刺激被重组PS_2蛋白免疫的小鼠脾细胞和B 淋巴细胞发生记忆性增殖并产生S1 多肽特异性抗体,促进小鼠产生IL-6,并可诱导未致敏人PBMC 增殖和B 细胞活化。结果提示,重组PS_2 蛋白可能诱生SARS-CoV 的中和抗体,因而成为一种新型的抗SARS 疫苗。
[Abstract]:2002 ~ 2003 outbreak of infectious atypical pneumonia (SARS) caused a total of 8098 cases of infection in 29 countries, 774 cases of death, caused widespread concern in the.SAR.S associated coronavirus (SAR.S.CoV) SARS pathogens. Spike protein (S protein) is the shell component of SARS - CoV, SARS-CoV and combined with the key structure of host recognition the cells of the neutralizing antibody could induce SAR.S.CoV, which can be used as a target for anti drug and vaccine research of SARS.CoV.
In order to develop a vaccine to prevent SARS diseases, we used bioinformatics methods for the prediction of protein structure and characteristic of S, it may contain B lymphocyte epitope peptides selected for 168aa-216aa (S_2) and 726aa.843aa peptide (S_6). Based on this, to construct the S_2 gene six dimer (PS_2 gene) and molecular chaperone 10-s6 fusion gene (C86 gene), using E.coli expressed recombinant PS_2 and CS_6 protein, and Ni2+ affinity chromatography method to obtain the purified recombinant protein. The recombinant PS_2 protein with IgGl in high titer antibody in mice induced by S1 subunit SARS-CoV, identify the full-length S protein peptide (S1 peptide) and inactivated SARS CoV. in addition, the recombinant PS_2 protein can stimulate in vitro by recombinant PS_2 protein immunized mice spleen cells and B lymphocytes and S1 peptide specific antibody memory proliferation, promote the production of mice IL-6 can induce PBMC proliferation and B cell activation in non sensitized humans. The results suggest that recombinant PS_2 protein may induce neutralizing antibodies in SARS-CoV, and thus become a new anti SARS vaccine.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392

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