Mechanism of immuno injury on inner ear autoimmune disease

内淋巴囊二次免疫反应后1天，耳蜗和前庭多个部位可见iNOS表达。此外，通过免疫组化表达ss-DNA、CAD和CPP32，可见螺旋韧带和血管纹发生凋亡变性。用牛内耳原抗原反复致敏后，小鼠产生的抗体能反应牛内耳以及小鼠肾、脑、肺和肝脏的几种蛋白质。为了研究纯化的内耳分离蛋白的抗原性，将粗内耳抗原分离成14个部分，用Mini全凝胶洗脱器洗脱。55 ~ 65 kDa蛋白组分单致敏诱导的内耳浸润细胞数量最多。用全凝胶洗脱获得了大量的内耳蛋白。54 ~ 62kDa内耳蛋白片段反复致敏产生的抗体与60 ~ 70kda牛内耳蛋白呈单条带，而与小鼠内耳蛋白无反应。IgG和C3补体定位于血管纹的血管。这些结果提示自身免疫性内耳疾病是由内耳特异性抗原引起的。进一步研究内耳特异性致病性抗原是听力动力学和病理研究的必要条件。此外，内耳特异性致病性候选蛋白的蛋白质组学分析将是确定蛋白质发生的必要条件。

Expression of iNOS was seen in several sites of the cochlea and vestibule one day following secondary immune reaction in the endolymphatic sac. Furthermore, apoptotic degeneration was seen in the spiral ligament and stria vascularis by immunohistochemical expression of ss-DNA, CAD and CPP32. Following recurrent sensitization with crude inner ear antigens of bovine, mouse produced antibodies that reacted several proteins of bovine inner ear as well as mouse kidney, brain, lung and liver. In order to investigate antigenicity of purified inner ear fractioned proteins, crude inner ear antigens were separated into 14 fractions eluted on Mini Whole Gel Eluter. Single sensitization with 55-65 kDa proteins fraction induced the highest number of inner ear infiltrating cells among the fractions. Large volume of fractioned inner ear proteins was obtained by Whole Gel Eluter. Recurrent sensitization off 54〜62kDa inner ear proteins fraction produced antibody that showed single bands with 60-70kDa of bovine inner ear proteins, but not reacted to proteins of mouse those organs. IgG and C3 complement localized in the vessels of the stria vascularis. These results suggest that autoimmune inner ear disease is caused by inner ear specific antigens. Hearing kinetics as well as pathological study is further necessary to investigate specific pathogenic inner ear antigen. Furthermore, proteome analysis of the candidates of inner ear specific pathogenic protein will be essential to identify protein genesis.