豚鼠自身免疫性梅尼埃病模型的主要内耳抗原分析

Analysis of the main components of inner ear antigens inducing autoimmune Meniere＇ s disease in guinea pigs

目的探寻导致豚鼠自身免疫性梅尼埃病（autoimmune Meniere＇s disease，AIMD）的主要内耳组织抗原成分。方法采用同种粗制内耳抗原免疫豚鼠，观察听觉功能、前庭功能及内耳组织形态学方面的变化，判断AIMD模型与非模型动物。采用免疫印迹法（Western blotting）对比分析模型动物与非模型动物血清内针对内耳组织抗原不同成分特异性免疫反应的差异，寻找只针对AIMD模型动物的特异性成分。结果内耳组织所含抗原成分较多，免疫后酶联免疫吸附试验（ELISA）结果显示，AIMD模型与非AIMD模型动物均不同程度地存在针对内耳组织抗原的血清抗体水平升高。听功能检测，非AIMD模型动物听力损失不明显。Western boltting结果显示，AIMD模型动物出现针对相对分子质量为68000、58000、42000及28000蛋白质成分的反应条带，而非AIMD模型动物则未显示这些条带的特异性抗原抗体反应。结论可能只有出现针对导致内耳自身免疫性疾病的主要抗原成分的特异性免疫反应，才会造成明显的内耳免疫病理损伤和功能障碍。相对分子质量为68000、58000、42000及28000的内耳组织抗原可能是导致豚鼠自身免疫性膜迷路积水的主要抗原成分。

Objective To investigate the main components of inner ear antigens inducing autoimmune Meniere＇s disease（AIMD） in guinea pigs. Methods The guinea pigs were immunized with isologous crude inner ear antigens （ICIEAg） . Then, the hearing function was measured with auditory brainstem response （ ABR）, the vestibular function was measured with electronystagmography （ including spontaneous nystagmus and caloric test）, and inner ear histopatholoical changes were observed by inner ear celloidin section with haematoxylin-eosin staining and observed under light microscope. According to these results, the AIMD-model animals from non-AIMD-model ones were distinguished. The special antibodies against ICIEAg in sera were measured with ELISA. The antigen-antibody reactions against different components of ICIEAg were detected by Western blotting with sera of AIMD and non-AIMD guinea pigs respectively. Then, we analysed the contrast between them and found the main components of the ICIEAg that were positive reaction in AIMD guinea pigs and negative reaction in non-AIMD guinea pigs. Results The result of ELISA demonstrated that the sera of both the AIMD and non-AIMD guniea pigs contained the special antibodies against ICIEAg after immunized with ICIEAg. The difference of the amount of antibody against ICIEAg between AIMD guinea pig group and non-AIMD guinea pig group was not significant. Western blotting assay showed only the sera of AIMD guinea pig contained the antibodies against the specific antigens with the molecular of 68 000, 58 000, 42 000 and 28 000. Conclusions ICIEAg contain many different components, the AIMD might only happen in the guinea pigs in which the special immunization against the main components that could induce this kind of disorder appeared . The inner ear antigens with molecular of 68 000, 58 000, 42 000 and 28 000 might be the main components inducing AIMD in guinea pigs.