三种内耳抗原与自身免疫性内耳病的相关性研究

Relationship between three inner ear antigens and autoimmune inner ear disease

目的 探讨 3种纯化的内耳抗原与自身免疫性内耳病的关系 ,并确定其在耳蜗的表达。方法 以粗制内耳抗原的 3种亚组份 (310 0 0、42 0 0 0～ 45 0 0 0和 6 0 0 0 0蛋白 )作为抗原 ,分别免疫动物(B、C和D组 ) ,观察听阈、血清IgG水平和内耳形态学的改变 ,并应用免疫组织化学方法确定其在耳蜗的表达。A组为对照组 ,以不含抗原的聚丙烯酰胺凝胶匀浆液代替内耳抗原。结果 免疫前各组听阈差异无显著性 (F =0 .0 7,P >0 .0 5 ) ,免疫后对照组和C组听阈和内耳形态学未见明显改变 ,B组 (9/ 30只 )和D组 (7/ 2 8只 )部分动物出现听力损失。组间听阈差异有显著性 (F =9.12 ,P <0 .0 1)。实验组动物血清IgG均显著性升高 (F =7.46 ,P <0 .0 1) ,B、C和D组与对照组相比差异有显著性。310 0 0蛋白完全分布于耳蜗神经纤维 ,而 42 0 0 0～ 45 0 0 0和 6 0 0 0 0蛋白分布广泛 ,螺旋神经节、Corti器、血管纹和螺旋韧带均有分布。结论 粗制内耳抗原中 310 0 0和 6 0 0 0 0亚组份均能独立诱发自身免疫性内耳病 ,310 0 0蛋白具有很高的组织特异性 。

Objective To investigate the relationship between three purified inner ear antigens and autoimmnne inner ear disease (AIED), and to determine their expressions in normal guinea pig cochleas. Methods Guinea pigs were divided into group B, group C and group D and immunized respectively with three subcomponents of crude inner ear antigens (31 000, 42 000～45 000 and 60 000 proteins ). Hearing thresholds, serum IgG levels and morphological changes of inner ear were observed. The protein expressions of these antigens were examined by immunohistochemistry. In the control group, antigens were replaced by gel homogenate of polyacrylamide. Results There were no significant differences of hearing threshold among groups before immunization with inner ear antigens( F =0.07, P >0.05).There were no significant changes of hearing thresholds and inner ear morphology in group C and control group. Hearing thresholds dropped in part of animals immunized with 31 000 or 60 000 protein. Hearing threshold of group B and D changed significantly after immunization, as compared with the control group( F =9.12, P <0.01). Serum IgG levels increased significantly in all experimental groups compared with the control group( F =7.46, P <0.01). The 31 000 protein distributed strictly in cochlear nerve, and 42 000 ～45 000 or 60 000 protein distributed widely, including the spiral ganglion, Corti's organ, stria vascularis and spiral ligament. Conclusions Two subcomponents of 31 000 and 60 000 in crude inner ear antigen could induce autoimmune inner ear disease. The distribution of 31 000 protein was more tissue specific and might be used as a marker protein for clinical diagnosis of autoimmune inner ear disease.