热休克蛋白70及其抗体与实验性自身免疫性内耳病

The role of heat shock protein 70 and its autoantibody in experimental autoimmune inner ear disease

目的 了解抗原免疫对热休克蛋白 70 (heatshockprotein 70 ,HSP70 )在豚鼠耳蜗中表达的影响。方法 以同种异体内耳组织为抗原免疫豚鼠 ,用免疫组织化学方法检测豚鼠耳蜗中HSP70的表达并进行计算机图像处理 ;对豚鼠膜迷路组织进行免疫转印分析以测定HSP71 ;以纯化的人类重组HSP71为抗原 ,用免疫转印技术 (Westernblot)检测动物血浆中HSP71抗体。结果 HSP70在正常豚鼠耳蜗中呈低水平表达。抗原免疫后 ,实验组动物耳蜗HSP70免疫活性较对照组明显增强 ,两组间的吸光度值差异有显著性 (P <0 0 1 ) ;免疫后大多数动物 (1 2 / 2 0 )体内产生了抗HSP71抗体 ,实验组HSP71抗体阳性率显著高于对照组 (P <0 0 1 )。结论 同种膜迷路组织作为抗原免疫豚鼠能够诱导HSP70在耳蜗内合成增加 ;表明HSP70与自身免疫性内耳病有一定联系。

Objective To examine the effect of antigen stimulation on induction of the heat shock protein 70(HSP70) in the guinea pig cochlea and to explore the role of HSP70 and its autoantibody in the pathogenesis and diagnosis of autoimmunize inner ear disease (AIED) Methods To establish the animal model of AIED, homologous crude inner ear antigen was used to immunize the guinea pig The expression of HSP70 was detected by immunocytochemistry and the relative staining densities were quantified with a light microscope image analysis system The autoantibodies against HSP71 in the plasma of animals were tested by Western blot using purified recombinant human HSP71 as antigen Results HSP70 presented in the normal guinea pig cochlea at a lower level After immunological challenge, the levels of HSP70 immunoreactivity in the immunized animals were significantly increased as compared with control animals Optical densities of cochleae of immunized animals were significantly greater than those of animals in the control group ( P <0 01) Most of the immunized animals had developed autoantibodies against HSP71 The incidence of autoantibody in the experimental group was significantly different ( P <0 01) from the control group Conclusion Antigen stimulation could lead to an increase in the synthesis of HSP70 in the guinea pig cochlea The detection of autoantibody against HSP71 might have significance in the diagnosis of AIED