旋毛虫DNA疫苗诱导小鼠免疫应答的研究

Immune response induced by DNA vaccine of Trichinella spiralis in BALB/c mice

目的 观察旋毛虫DNA疫苗在小鼠体内诱导的免疫应答。方法 将旋毛虫肌幼虫编码 31kDa抗原结构基因真核表达质粒pcDNA3-TspE1分别用肌肉注射和基因枪免疫BALB/c小鼠 ,免疫血清用旋毛虫肌幼虫冰冻切片及石蜡切片抗原进行IFAT检测 ,并用旋毛虫肌幼虫可溶性抗原进行Westernblot分析。结果 IFAT表明 pcDNA3-TspE1接种BALB/c小鼠后产生的免疫血清与旋毛虫肌幼虫可产生阳性反应 ,在肌幼虫冰冻及石蜡切片上均显示黄绿色荧光。Westernblot检测结果显示 ,pcDNA3-TspE1接种小鼠后产生的免疫血清只能识别旋毛虫肌幼虫可溶性抗原中的 31kDa抗原组分 ,而 pcDNA3质粒接种小鼠后的血清不能与旋毛虫肌幼虫可溶性抗原发生免疫反应。结论 旋毛虫DNA疫苗 pcDNA3-TspE1能够诱导小鼠产生特异性体液免疫应答。

Objective To observe the immune response of mice induced by DNA vaccine of Trichinella spiralis.Methods The BALB/c mice were immunized with the recombinant eukaryotic expression plasmid pcDNA3-TspE1 contained the structural gene encoding a 31 kDa antigen of Trichinella spiralis through two routes: intramuscle injection and gene-gun injection.The anti-Trichinella antibodies in immune sera were detected by indirect fluorescencent antibody test(IFAT)with freezing section of T.spiralis muscle larvae and paraffin section of muscle containing T.spiralis larvae as antigen.The immune sera were also analized by Western blot using T.spiralis muscle larval soluble antigens.Results The results of IFAT showed that the immune sera from mice inoculated by pcDNA3-TspE1 could produce the positive reaction with T.spiralis muscle larvae,the yellow-green fluorescence appeared on the freezing and paraffin section of T.spiralis larvae.Western blot confirmed that sera from mice immunized by pcDNA3-TspE1 were only reacted with the 31kDa antigenic component of T.spiralis larval sobuble antigen,whereas no positive immune reaction was observed between the sera from mice inoculated with pcDNA3 and the soluble antigen of T.spiralis muscle larvae.Conclusion The specific humoral immune response in mice could be produced by DNA vaccine(pcDNA3-TspE1)of Trichinella spiralis.