ANTIGENIC TARGETS AUTOIMMUNE SENSORINEURAL HEARING LOSS

描述：(改编自申请人的描述) 男性自身免疫性感音神经性耳聋可能是 针对内耳抗原的抗体。与KHRI-3结合的单抗 Corti器中的支持细胞和豚鼠内脏的68kD蛋白 耳提取物导致豚鼠听力丧失和外毛细胞丧失 猪。在50-60%的AISNHL患者的血清中，我们发现了与 与KHRI-3相似。我们将检验AISNHL患者的假设 有对68kD内耳抗原的抗体，该抗原在其上表达 科尔蒂器官中的支持细胞，同样的抗原也 在人类内耳中表达。我们将检验第二个假设，即人类 自身抗体和KHRI-3结合到相同的内耳蛋白上，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柱上纯化的亲和力，测序，这一基因 蛋白质可以被克隆、测序、体外表达并用作抗原 检测AISNHL患者抗体的底物。初步结果 显示具有68kD豚鼠内耳抗原抗体的人血清 也可与豚鼠内耳的支持细胞结合。人类内耳 组织可以特异性地吸收这种抗体反应性。68kD蛋白 KHRI-3从豚鼠内耳提取液中的免疫沉淀 AISNHL血清，但不是正常捐赠者的血清，也不是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层析柱将分离出一种 氨基酸测序需要足够的纯度。根据我们将列出的序列 筛选内耳文库，分离和克隆内耳蛋白基因， 在体外翻译系统中表达全长c DNA，并进行检测 该材料能否作为检测AISNHL的底物 抗体。如果我们成功了，这可能会导致一次合理、准确的测试 对于AISNHL和更好地理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.