抗猪带绦虫六钩蚴兔血清的制备及重组质粒pcDNA3.1/TSO45-4B在小鼠骨骼肌的表达

徐胜贵; 方强; 孙新; 夏惠; 杨小迪; 胡守锋; 陈兴智; 王雪梅

抗猪带绦虫六钩蚴兔血清的制备及重组质粒pcDNA3.1/TSO45-4B在小鼠骨骼肌的表达

Preparation of rabbit sera of anti-Taenia solium onchosphere and in vivo expression of recombinant plasmid pcDNA3.1/TSO45-4B in skeletal muscle of vaccinated mice

摘要

摘要: 目的: 制备抗六钩蚴全虫可溶性抗原的兔血清,观察本室制备的猪带绦虫六钩蚴期特异性抗原基因TSO45-4B的重组核酸疫苗pcDNA3.1/TSO45-4B经肌肉接种后在宿主体内的表达。方法: 用次氯酸钠法孵化六钩蚴,Dot-Blot、ELISA检测兔血清中特异性抗体及滴度;以pcDNA3.1/TSO45-4B进行体内转染,继之免疫组化法观察其在鼠体骨骼肌的表达。结果: 抗六钩蚴兔血清多克隆抗体制备成功,抗体效价为1:20 100;免疫组化染色后在重组质粒注射部位肌肉组织的肌纤维内显示阳性的棕黄色分泌颗粒。结论: 重组质粒在小鼠骨骼肌内成功地表达为猪囊尾蚴病多基因DNA疫苗的研制提供了体内表达的依据之一。

Abstract: Objective: To prepare anti-onchosphere rabbit sera and observe the in vivo expression of recombinant plasmid pcDNA3.1/TSO45-4B containing gene of stage-specific antigen TSO45-4B of Taenia solium onchosphere.Methods: The soluble antigen of onchosphere collected from hatched eggs was injected into rabbits,and the anti-onchosphere rabbit sera were collected and detected by Dot-Blot and ELISA.The recombinant plasmid pcDNA3.1/TSO45-4B was injected into Kunming mice's skeletal muscle and its expression was detected by immunohistochemistry assay.Results: The titer of polyclonal antibody to antigen of onchosphere in rabbit serum was(1:20 100).The specific masculine brown-yellow color reaction was seen in the muscle fibers of recombinant plasmid injected group.Conclusions: The successful expression of pcDNA3.1/TSO45-4B has provided the basis of in vivo expression for developing DNA vaccine for prevention of cysticercosis.

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