ANTIGENIC TARGETS AUTOIMMUNE SENSORINEURAL HEARING LOSS

描述：（改编自申请人的描述） 人类自身免疫性感音神经性听力损失（AISNHL）可能是以下原因的结果： 内耳抗原的抗体一种单克隆抗体KHRI-3， Corti器支持细胞和豚鼠内耳中一种68 kD蛋白 耳提取物导致豚鼠听力损失和外毛细胞损失 猪在50-60%的AISNHL患者的血清中，我们发现了具有以下特征的抗体： 与KHRI-3相似。我们将检验AISNHL患者 具有针对68 kD内耳抗原的抗体，该抗原表达于 支持细胞在Corti器官，而且同样的抗原也是 在人的内耳中表达。我们将测试第二个假设， 自身抗体和KHRI-3结合相同的内耳蛋白，这种蛋白 与热休克蛋白70（HSP-70）不同，内部蛋白可以是 在KHRI-3柱上亲和纯化，测序，并且该基因 蛋白质可被克隆、测序、体外表达并用作抗原 用于检测患有AISNHL的患者中的抗体的底物。初步结果 显示具有68 kD豚鼠内耳抗原抗体的人血清 也与豚鼠内耳的支持细胞结合。人内耳 组织可以特异性地吸收这种抗体反应性。A 68 kD蛋白 从豚鼠内耳提取物中通过KHRI-3免疫沉淀的细胞被染色， AISNHL血清，但不能通过正常供体的血清和HSP-70的抗体。 这些初步数据支持我们的假设，并表明， 用KHRI-3抗体制备的免疫亲和柱将分离 纯度足以进行氨基酸测序。根据序列，我们将 筛选内耳文库，分离并克隆内耳蛋白的基因， 在体外翻译系统中表达全长cDNA，并测试 该材料是否可用作检测AISNHL的底物 抗体的如果我们成功了，这可能会导致一个合理的，准确的测试， 和更好地了解自身免疫损伤的机制， 内耳

DESCRIPTION: (Adapted from applicant's description) Autoimmune sensorineural hearing loss (AISNHL) in man may be the result of antibodies to inner ear antigens. A monoclonal antibody KHRI-3 that binds to supporting cells in the organ of Corti and a 68 kD protein in guinea pig inner ear extracts causes hearing loss and loss of outer hair cells in the guinea pig. In sera from 50-60% of patients with AISNHL we found antibodies with similarities to KHRI-3. We will test the hypothesis that patients with AISNHL have antibody to a 68 kD inner ear antigen, that this antigen is expressed on supporting cells in the organ of Corti, and that the same antigen is also expressed in human inner ear. We will test a second hypothesis that human autoantibodies and KHRI-3 bind to the same inner ear protein, that this protein is distinct from heat shock protein 70 (HSP-70), that the inner protein can be affinity purified on a KHRI-3 column, sequenced, and that the gene for this protein can be cloned, sequenced, expressed in vitro and used as an antigenic substrate for detecting antibodies in patients with AISNHL. Preliminary results show that human sera with antibodies to a 68 kD guinea pig inner ear antigen also bind to supporting cells in the guinea pig inner ear. Human inner ear tissue can specifically absorb this antibody reactivity. A 68 kD protein immunoprecipitated from guinea pig inner ear extracts by KHRI-3 is stained by AISNHL sera, but not by normal donors' sera and not by antibodies to HSP-70. These preliminary data support our hypotheses and suggest that an immunoaffinity column prepared with KHRI-3 antibodies will isolate a protein of sufficient purity for amino acid sequencing. Based on the sequence we will screen inner ear libraries, isolate and clone the gene for the inner protein, express the full length cDNA in an in vitro translation system, and test whether this material can be used as a substrate for detecting AISNHL antibodies. If we are successful this could lead to a rational, accurate test for AISNHL and a better understanding of the mechanism of autoimmune damage to the inner ear.