三种内耳抗原与自身免疫性内耳病的相关性研究

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(31000、42000～45000和60000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=0.07,P>0.05),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(7/28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<0.01).实验组动物血清IgG均显著性升高(F=7.46,P<0.01),B、C和D组与对照组相比差异有显著性.31000蛋白完全分布于耳蜗神经纤维,而42000～45000和60000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31000和60000亚组份均能独立诱发自身免疫性内耳病,31000蛋白具有很高的组织特异性,可能作为一种标志性蛋白用于自身免疫性内耳病的临床诊断.     

内耳抗原和自身免疫性内耳病

自身免疫性内耳病是少数几种经过恰当治疗后可获得较好疗效的内耳病之一，但由于其发病机理仍不清楚，特别是缺乏高度特异性的实验室检查方法，使得诊断较为困难。严格来说，确诊应有病理结果，内耳的不可活检性增加了本病诊断的难度，因此寻求内耳特异性抗原成为目前急需解决的课题。本就近年来新发现的数种内耳抗原，对它们的发现经过以及相关的动物实验和临床研究作了介绍，以期为寻找内耳特异性抗原提供新的思路。     

内耳抗原和自身免疫性内耳病

自身免疫性内耳病是少数几种经过恰当治疗后可获得较好疗效的内耳病之一,但由于其发病机理仍不清楚,特别是缺乏高度特异性的实验室检查方法,使得诊断较为困难.严格来说,确诊应有病理结果,内耳的不可活检性增加了本病诊断的难度,因此寻求内耳特异性抗原成为目前急需解决的课题.本文就近年来新发现的数种内耳抗原,对它们的发现经过以及相关的动物实验和临床研究作了介绍,以期为寻找内耳特异性抗原提供新的思路.     

P0蛋白的分布及其与自身免疫性内耳病的关系

P0蛋白是周围神经系统髓鞘形成细胞Schwann细胞膜上的一种结构蛋白，对维持髓鞘正常的结构和功能有十分重要的意义。资料表明P0蛋白还与周围神经系统及内耳自身免疫病的发生有关。本就P0蛋白的结构和功能、分布及其与自身免疫性疾病的关系作一综述。     

自身免疫性内耳病的实验研究

用同种异体内耳抗原免疫豚鼠，观察听阈、血清免疫学、形态学及免疫病理学的改变，结果发现部分豚鼠ＡＰ（Ｎ＿１）反应阈明显升高、血清中出现抗内耳自身抗体，且表现有膜迷路积水、蜗轴血管炎、螺旋神经元细胞空泡变性、细胞数减少，特别是部分动物蜗轴血管和血管纹毛细血管内皮有ＩｇＧ沉积。提示通过这种实验方法可在部分动物成功地建立实验性自身免疫性内耳病动物模型。此外，就该模型与该病临床表现的相似性进行了比较。     

不同内耳组织抗原免疫致自身免疫性感音神经性聋的研究

目的：探讨不同内耳组织抗原免疫所致内耳主要病理损伤部位和听力障碍类型。方法：采用同种螺旋韧带（ＳＬ）、基底膜（ＢＭ）、螺旋神经节（ＳＧ）组织抗原免疫豚鼠，观察内耳组织病理改变和听觉功能变化。结果：ＳＬＡｇ和ＢＭＡｇ免疫组主要表现耳蜗微音器电位阈值升高和复聪现象，以及蜗管内和血管纹的免疫炎性病理改变；ＳＧＡｇ免疫组主要表现听神经复合动作电位阈值升高和幅值降低，内耳病理变化主要位于蜗轴血管及周围和ＳＧ     

一种筛选自身免疫性疾病患者抗内耳抗体的新方法

目的 探讨用一种新建立的外加电场固定液相分子快速斑点免疫分析（ｒａｐｉｄ ｅｌｅｃｔｒｉｃ ｆｉｅｌｄ ｉｍｍｏｂｉｌｉｚｉｎｇ ｌｉｑｕｉｄ ｐｈａｓｅ ｍｏｌｅｃｕｌｅ ｄｏｔ ｂｌｏｔ,ＲＥⅡＭＤ）法检测自知免疫性疾病患者抗体的临床意义。方法 收集１１５中确诊的自身免疫性疾病患者共７１例,突发性聋２１例,非免疫性疾病患者４８例（对照组）。检测内容包括一般免疫指标及自身抗体如抗ｓｍｏｏｔｈ ｍ     

Relationship between three inner ear antigens with different molecular weights and autoimmune inner ear disease

Crude inner ear antigen (CIEAg) can induce autoimmune inner ear disease (AIED) although it is not known which subcomponent of CIEAg is involved. In this study, we investigated the relationship between 3 purified inner ear antigens (31, 42-45 and 60 kD proteins) and AIED, and determined their distribution in normal guinea pig cochlea. Three groups of guinea pigs were immunized with the three inner ear antigens and one group served as a control. The hearing thresholds, serum IgG level and morphological changes in the inner ear were observed. The expression of the three antigens in the cochlea was detected using immunohistochemical techniques. No obvious changes in hearing thresholds or inner ear morphology were observed between the control and 42-45 kD groups. Animals immunized with the 31 or 60 kD proteins showed a significant increase in hearing thresholds (p < 0.05 vs control), accompanied by morphological changes in the inner ear. The serum IgG level was increased significantly (p < 0.05) in all immunized animals. The 31 kD protein was distributed in the cochlear nerve and spiral ganglion, while the 42-45 and 60 kD proteins were distributed widely, being found in the spiral ganglion, organ of Corti, stria vascularis and spiral ligament. These results suggest that two subcomponents of CIEAg (the 31 and 60 kD proteins) may induce AIED independently, that several inner ear antigens may contribute to the pathogenesis of AIED and that the 31 kD protein is of high tissue specificity and may be used as a marker protein for the clinical diagnosis of AIED.     

Experimental autoimmune inner ear disease

A guinea pig animal model of autoimmune inner ear disease is presented. The functional, anatomical, and immunological inner ear changes were tested electrophysiologically, histologically and by the immunofluorescence test. Using a homologous crude inner ear antigen (CIEAg) we were able to induce endolymphatic hydrops, vasculitis, mild cellular infiltration of the endolymphatic sac, and occasional spiral ganglion degeneration. Threshold shift was seen in 20% of the tested ears. Specific fluorescence was revealed around the modiolar vessels and in the basilar membrane. The endolymphatic sac and duct showed occasional fluorescence in the epithelial and/or subepithelial layers. The findings were discussed in light of the other models immunized with various forms of inner ear antigens. Similarities between the detected specific fluorescence and the fluorescence revealed by sera of patients with cochleovestibular disorders were discussed.     

自身免疫性内耳病动物模型的建立

目的建立一种自身免疫性内耳病的动物模型，其具有可重复性高，适于进行深入免疫学分析的特点。方法提取豚鼠内耳膜迷路组织为抗原，与等量完全弗氏佐剂，百日咳杆菌一次免疫C57BL／6小鼠。检测反应阈、血清免疫学、内耳形态学及免疫组织化学的改变。结果免疫后小鼠听性脑干反应阈显著提高，内耳中出现显著的炎性细胞浸润、内淋巴积水和螺旋神经节细胞变性、数量减少等形态学改变，血清中可检测到抗内耳自身抗体，鼓阶内浸润细胞中的淋巴细胞主要为CD4^ T细胞。结论应用这种方法在C57BL／6小鼠可成功建立自身免疫性内耳病的动物模型。     

自身免疫性内耳病的诊治进展

自身免疫性内耳病（autoimmune inner ear disease,AIED）是一组以内耳损害为主的免疫介导的疾病, 常以波动性单或双侧感音神经性聋、眩晕、耳鸣及耳胀满感为主要临床表现。虽然临床医师已经关注到AIED,但由于缺乏典型的临床特征及特异性的实验室检查,致该病在临床上存在较多的漏诊及误诊。近年来,随着内耳免疫学研究的深入,人们对AIED的认识也取得了较大提高,现就AIED的发病机制、诊断及治疗等方面的研究现状做一综述。     

Sudden hearing loss and autoimmune inner ear disease

This case report describes the audiologic and medical diagnostic evaluations, results, and treatment options in a patient with a classic presentation of immune-mediated sensorineural hearing loss, commonly called autoimmune inner ear disease (AIED). It reviews findings of the basic battery, immittance audiometry, transient otoacoustic emissions, and auditory brainstem response measures and medical findings over more than 2 years. AIED generally causes asymmetric bilateral sensorineural hearing loss with atypical configuration. Although hearing loss is generally fluctuant, the overall pattern is usually rapid progression, particularly in the absence of early medical intervention. Word recognition is usually disproportionately poor. In our case, otoacoustic emissions and auditory brainstem responses suggest both cochlear and retrocochlear involvement and may initially appear to be inconsistent with pure-tone thresholds. Audiologists must be familiar with AIED because early identification is critical. Additionally, an immunologic basis may be a factor in other disorders, including many cases of Meniere's disease.     

Experimental autoimmune lesions of the inner ear]

This paper presents the autoimmune lesions of the inner ear by using isologous inner ear antigen (IEAg) on the guinea pigs. The purposes of this study were to determine whether the inner ear is an immune response organ and what is the sequelae of the immune processes both on physiological and morphological changes in the inner ear. Animals were systemically sensitized with IEAg in complete Freund's adjuvant and boothed with IEAg in incomplete Freund's adjuvant three times in every 3-4 weeks and then allowed to survive 4-6 weeks. For the ABR measurement, 8 of 28 ears (28.6%) in the experimental group showed threshold enhancement greater than or equal to 10dB before the animal sacrifice and the mean threshold shift of the ABR 8 weeks post immunization and before sacrifice revealed significant difference. Histological specimens were processed for both light and electron microscopic study. In the LM study, it showed endolymphatic hydrops, local thickening of the Reissener's membrane, and spiral ganglion cells lost. In the SEM, disturbance and loss of the cilia of the outer hair cells of the cochlea and the type I and II sensory cells in the crista ampularis can be seen. Also, some otoconia showed numerous malformation.     

Clinical and experimental studies of autoimmune inner ear disease

Otolaryngologists have long sought to identify causes of sensorineural hearing loss that could be reversed by medical treatment. An increasing amount of clinical and experimental evidence indicates that this postulated entity is related to autoimmune inner ear disease. Because of the lack of well-defined detection methods for identifying autoimmune processes within the human inner ear and the fact that it is one of the few organs of the body not amenable to diagnostic biopsy there has been great interest in developing animal models that mimic clinical entities. Different models will be presented in this paper. The results of these studies should provide sufficient evidence to establish a clear position in mainstream immunology for the entity of autoimmune inner ear disease.     

豚鼠自身免疫性梅尼埃病模型的主要内耳抗原分析

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 bohting结果显示,AIMD模型动物出现针对相对分子质量为68 000、58 000、42 000及28 000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68 000、58 000、42 000及28 000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.     

自身免疫性内耳病动物模型的建立

目的　建立一种自身免疫性内耳病的动物模型 ,其具有可重复性高 ,适于进行深入免疫学分析的特点。方法　提取豚鼠内耳膜迷路组织为抗原 ,与等量完全弗氏佐剂 ,百日咳杆菌一次免疫C5 7BL/6小鼠。检测反应阈、血清免疫学、内耳形态学及免疫组织化学的改变。结果　免疫后小鼠听性脑干反应阈显著提高 ,内耳中出现显著的炎性细胞浸润、内淋巴积水和螺旋神经节细胞变性、数量减少等形态学改变 ,血清中可检测到抗内耳自身抗体 ,鼓阶内浸润细胞中的淋巴细胞主要为CD4 T细胞。结论　应用这种方法在C5 7BL/6小鼠可成功建立自身免疫性内耳病的动物模型