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目的探讨重组白细胞介素-4(IL-4)真核表达质粒增强日本血吸虫组织蛋白酶BDNA疫苗对小鼠的免疫保护效果。方法将小鼠IL-4基因克隆至真核表达载体pcDNA3.1以构建小鼠重组IL-4表达质粒,并联合日本血吸虫组织蛋白酶B的表达质粒DNA(VR1012-Sj31)肌注免疫小鼠,设重组IL-4表达质粒、组织蛋白酶B表达质粒和2种空载体质粒对照组。免疫组化检测IL-4和组织蛋白酶B在小鼠肌细胞内的表达,末次免疫3周后攻击感染,用减虫率及减卵率表示保护力。结果重组IL-4和组织蛋白酶BDNA均在小鼠肌细胞表达,重组IL-4和组织蛋白酶BDNA联合免疫小鼠,产生43.2%的减虫率和76.6%的减卵率,与组织蛋白酶BDNA单独免疫相比差异有显著性(P<0.01,P<0.05)。结论重组IL-4能提高日本血吸虫组织蛋白酶B核酸疫苗的抗血吸虫保护力作用,具有佐剂的免疫效应。
Objective To investigate the effect of recombinant interleukin-4 (IL-4) eukaryotic expression plasmid on the immunoprotection of Schistosoma japonicum cathepsin B DNA vaccine in mice. Methods The murine IL-4 gene was cloned into the eukaryotic expression vector pcDNA3.1 to construct mouse recombinant IL-4 expression plasmid and immunized with intramuscular injection of Schistosoma japonicum cathepsin B expression plasmid DNA (VR1012-Sj31) Recombinant IL-4 expression plasmid, cathepsin B expression plasmid and two empty vector plasmid control group. Immunohistochemistry was used to detect the expression of IL-4 and cathepsin B in mouse muscle cells. The infection was challenged 3 weeks after the last immunization, and the protection was indicated by the worm reduction rate and egg reduction rate. Results Recombinant IL-4 and cathepsin B DNA were all expressed in mouse muscle cells. Combined immunization with recombinant IL-4 and cathepsin B DNA resulted in a 43.2% reduction in worm rate and a 76.6% reduction in ovulation rate compared with cathepsin BDNA alone Immunity compared with the difference was significant (P <0.01, P <0.05). Conclusion Recombinant IL-4 can improve the anti-schistosomucosal protective effect of Schistosoma japonicum cathepsin B nucleic acid vaccine and has adjuvant immune effect.