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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。
Objective To detect the expression of recombinant adenovirus Ad-SN in vitro and in vivo and to investigate its humoral immune response induced in rats. Methods The expression of SN gene was detected by RT-PCR and Western blotting in Ad-SN-infected Vero-E6 cells. The expression of Ad-SN was detected by Western blot in subcutaneous tissue of rats. Ad-SN was subcutaneously immunized with anti-SARS-CoVIgG in serum. Results There was SN gene transcription in Vero-E6 cells infected with Ad-SN, and the mRNA level was in a dose-dependent manner. In the cells infected with Ad-SN, both the culture supernatant and the Ad-SN injection site tissues could be detected SN expression. High-titer SARS coronavirus antibody (IgG) was detected in the serum of Ad-SN immunized rats. Conclusions Ad-SN can express secreted SN protein and induce SARS-CoV specific humoral immune response in rats.