【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。更多还原

【Abstract】 To detect the expression of the N-terminal fragment of spike gene of SARS-CoV mediated by adenoviral vector Ad-S_N and to investigate the humoralimmunity induced by Ad-S_N in rats, Vero-E6 cells were infected with Ad-S_N at different dosages and the expression of the S_N gene in Vero-E6 cells was detected by RT-PCR. Rats were infected subcutaneously with Ad-S_N, and the expression of gene products in rats was investigated by Western blotting analysis. Rats were immunized with Ad-S_N 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 S_N gene could be detected in the Ad-S_N infected Vero-E6 cells in a dose-dependent manner with the amount of Ad-S_N, and the expression of S_N could be delected in the cell culture supematants, the Ad-S_N-infected cells and in the local tissues of rats injected with Ad-S_N. Also, high titers of the SARS-CoV-specific antibody IgG could be detected on the sera of rats immunized with Ad-S_N . It is concluded that Ad-S_N expresses the secretary type S_N protein of which the SARS-CoV-specific humoral immune responses can be induced in rats.更多还原