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目的 :用免疫组化方法对猪囊尾蚴抗原 Annexin32进行组织定位。 方法 :首先将 Annexin32在大肠杆菌中进行表达 ,表达的 Annexin32蛋白经亲和层析、凝血酶切割纯化 ,以囊虫病病猪血清检测表达蛋白的抗原性 ;然后将纯化蛋白免疫新西兰白兔 ,获得抗 Annexin32的多克隆抗体 ;最后将新鲜剥离的囊尾蚴进行冰冻切片 ,用免疫组化方法进行定位研究。 结果 :表达的 Annexin32蛋白可以被囊虫病病猪血清特异地识别。制备的多克隆抗体的效价为 1∶ 4 0万。免疫组化定位结果显示 ,Annexin32主要在囊壁中表达 ,头节部表达较少 ;囊尾蚴周边肌肉组织也有表达。 结论 :Annexin32主要分布于囊尾蚴的囊壁 ,而且它可能是一种分泌性蛋白 ,可以分泌到周围的肌肉组织中
OBJECTIVE: To characterize the annexin 32 of cysticercus by immunohistochemistry. METHODS: Annexin32 was first expressed in E. coli. The expressed Annexin32 protein was purified by affinity chromatography and thrombin cleavage. The antigenicity of the expressed protein was detected by the serum of cysticercosis pigs. The purified protein was then used to immunize New Zealand white rabbits, Anti-Annexin32 polyclonal antibodies were obtained. Finally, freshly-dissected Cysticercus cellulosae were frozen and sliced, and the immunohistochemical method was used for localization study. Results: The expressed Annexin32 protein could be specifically recognized by the serum of cysticercosis pigs. The prepared polyclonal antibody has a titer of 1: 400,000. Immunohistochemical localization results showed that Annexin32 was mainly expressed in the cystic wall, with less expression in the cranial ganglia; the muscle tissue around the cysticercus was also expressed. Conclusion: Annexin32 is mainly distributed on the cysticercus cyst wall, and it may be a secreted protein that can be secreted into the surrounding muscle tissue