Yeom Kristen, Gray J, Nair T S, Arts H A, Telian S A, Disher M J, El-Kashlan H, Sataloff R T, Fisher S G, Carey T E
Immunology and Cell Biology Laboratory, Kresge Hearing Research Institute, Department of Otolaryngology-Head and Neck Surgery, The University of Michigan School of Medicine, 1301 East Ann Street, Ann Arbor, MI 48109-0506, USA.
Laryngoscope. 2003 Oct;113(10):1770-6. doi: 10.1097/00005537-200310000-00020.
To evaluate serum antibody to heat shock protein (HSP) 70 as a marker for autoimmune sensorineural hearing loss (AISNHL).
Sera from 20 patients with rapidly progressive sensorineural HL and 20 control volunteers without HL were tested for antibody reactivity against multiple HSP 70 substrates. Substrates included recombinant human HSP (rHuHSP) 72, purified bovine brain heat shock cognate (HSC) 73 and HSP 72, as well as heat-shocked and non-heat-shocked protein extracts from bovine kidney (MDBK) cells. All serum donors were previously tested for antibody to guinea pig inner ear supporting cells; 17 of 20 patients but none (0 of 20) of the controls were positive.
Sera were tested using Western blots.
Reactivity with rHuHSP 70 was observed in 16 patients and 17 controls. Similarly, 15 of 20 patients and 17 of 20 controls stained for both HSP 72 and HSC 73 from the bovine brain. When tested against the heat-shock-induced and control MDBK extracts, six patients and nine controls had greater reactivity with the induced HSP 72.
The frequency of antibodies to HSP substrates did not differ in patients and controls. Prior studies reported that HSP 72 is the 68 kD antigen commonly detected by AISNHL sera. However, we show that HSP 72 antibodies are no more prevalent in patients than in normal controls. Thus, it is unlikely that the 68 kD protein is HSP 72. Therefore, HSPs are not appropriate substrates for serodiagnosis of AISNHL.
评估血清热休克蛋白（HSP）70抗体作为自身免疫性感音神经性听力损失（AISNHL）标志物的价值。
检测20例快速进展性感音神经性听力损失患者及20名无听力损失的对照志愿者血清针对多种HSP 70底物的抗体反应性。底物包括重组人HSP（rHuHSP）72、纯化的牛脑热休克同源蛋白（HSC）73和HSP 72，以及来自牛肾（MDBK）细胞的热休克和非热休克蛋白提取物。所有血清供体之前均检测过针对豚鼠内耳支持细胞的抗体；20例患者中有17例呈阳性，而20名对照者均为阴性（0例）。
采用蛋白质免疫印迹法检测血清。
16例患者和17名对照者的血清与rHuHSP 70有反应性。同样，20例患者中有15例、20名对照者中有17名对来自牛脑的HSP 72和HSC 73均呈染色阳性。当检测热休克诱导的和对照MDBK提取物时，6例患者和9名对照者对诱导的HSP 72反应性更强。
患者和对照者中针对HSP底物的抗体频率无差异。既往研究报道HSP 72是AISNHL血清中常见检测到的68 kD抗原。然而，我们发现HSP 72抗体在患者中的流行率并不高于正常对照者。因此，68 kD蛋白不太可能是HSP 72。所以，HSP不适用于AISNHL的血清学诊断。
