腺病毒介导SARS病毒SN基因表达及其诱导的体液免疫  

Adenovirus-mediated expression of the N-terminal fragment of spike gene of SARS Coroarvirus and induction of specific humoral immunity

作　　者：刘然义[1] 黄嘉凌[1] 吴立志[1] 黄必军[1] 柯妙拉[1] 张艳玲[1] 王俊梅[2] 黄文林[1] 

机构地区：[1]中山大学肿瘤防治中心,广州510060 [2]中山大学基础医学院组胚教研室,广州510089

出　　处：《中国人兽共患病杂志》2005年第8期670-673,共4页Chinese Journal of Zoonoses

基　　金：国家“973计划"SARS研究专项(2003CBS14107);教育部抗非典科技攻关项目;广东省抗非典科技攻关项目;广州市科技攻关项目(2003Z3-E0451);深圳键康元药业集团有限公司资助。

摘　　要：目的检测重组腺病毒Ad-SN在体内外的表达,初步探讨其在大鼠内诱导的体液免疫反应。方法Ad-SN感染Vero-E6细胞后用RT-PCR和WesternBlot法检测SN基因的表达;用WesternBlot法检测Ad-SN在大鼠皮下组织中的表达。Ad-SN皮下免疫大鼠,用ELISA法测定血清中抗SARS-CoVIgG水平。结果在Ad-SN感染的Vero-E6细胞中存在SN基因的转录,其mRNA量呈剂量依赖性;在Ad-SN感染的细胞、其培养上清及Ad-SN注射部位组织中均能检测到SN的表达。在Ad-SN免疫大鼠血清中可检测到高滴度SARS冠状病毒抗体(IgG)。结论Ad-SN能表达分泌型SN蛋白,并在大鼠体内诱导SARS-CoV特异的体液免疫反应。To detect the expression of the N-terminal fragment of spike gene of SARS-CoV mediated by adenoviral vector Ad-SN and to investigate the humoralimmunity induced by Ad-SN in rats, Veto-E6 cells were infected with Ad-SN at different dosages and the expression of the SN gene in Veto-E6 cells was detected by RT-PCR. Rats were infected subcutaneously with Ad-SN, and the expression of gene products in rats was investigated by Western blotting analysis. Rats were immunized with Ad-SN at a dosage of 1 x 10^7 pfu/dose once a week for 3 consecutive weeks. Sera were taken from the immunized rats 2 weeks after immunization and the SARS-CoV-specific antibody IgG was detected by ELISA assay. It was found that the transcription of the SN gene could be detected in the Ad-SN infected Vero-E6 ceils in a doee-dependent manner with the amount of Ad-SN, and the expression of SN could be ddected in the cell culture supematants, the Ad-SN-infected ceils and in the local tissues of rats injected with Ad-SN. Also, high titers of the SARS-CoV-specific antibody IgG could be detected on the sera of rats immunized with Ad-SN. It is concluded that Ad-SN expresses the secretary type SN protein of which the SARS-CoV-specific humoral immune responses can be induced in rats.

关 键 词：疫苗 SARS冠状病毒 腺病毒载体 刺突蛋白 体液免疫 

分 类 号：R373.9[医药卫生—病原生物学]