目的探讨树突状细胞(DCs)是否及通过何种途径参与内耳的免疫应答,分析中耳炎免疫引发的内耳免疫对内耳功能的影响。方法将Hoechst33342标记的DCs分别经由脑脊液,颈外静脉及颈部皮下注入豚鼠体内,并在无菌条件下于右耳经鼓膜注射1×10^8·L^-1的金葡菌液100μL,左耳作正常对照,造急性中耳炎豚鼠模型。3天后基底膜切片及铺片若单明一鬼笔环肽(Phalloidin)染色,荧光显微镜(Olympus)及共聚焦显微镜观察。结果三种途径注入的DCs均可以在中耳炎豚鼠的中耳粘膜发现。在豚鼠内耳前庭阶、鼓阶内可以发现大量的免疫细胞渗入,同时可见DC位于内耳,数目很少,分布于基底膜、血管纹、螺旋神经节及壶腹嵴。三组动物对照耳仅皮下注射组有一例在耳蜗发现DCs渗入。器官切片除脑脊液组在脑中发现DC外,其余均未见DCs渗入。结论DCs可以通过不同途径参与中耳炎所致的内耳免疫反应。作为最强的抗原提呈细胞,免疫反应的启动者,DCs在内耳诱导强烈的免疫反应,这种免疫反应可能对内耳的结构和功能产生不可逆的严重损伤,DCs功能紊乱可能为自身免疫性内耳疾病的原因之一。提示抑制过度免疫反应能够有效保护内耳的免疫损伤。
Objective To study the influence on morphology and function by DCs injected into the inner ear via different routes in guinea pigs with otitis media.Methods: DCs labeled with Hoechst33342 were injected through the cerebrospinal fluid, the external jugular vein and neck skin respectively in guinea pigs. The middle ear cavity on right side was inoculated with 0.1ml 1×10^8/ml staphloccoccosis aureus, with the left ear serving as the control. Changes in the cochlea were examined 3 days after infection by fluorescence microscopy and confocal microscopy. Results DCs injected via all three routes were found in the middle ear mucosa in these guinea pigs with otitis media. DCs also were seen in the base- ment membrane, spiral ligament, stria vascularis, spiral ganglion and crista ampullaris, but the number was extremely low. In only one case DCs infiltration was seen in the cochlea on the control side. DCs infiltration in the brain was seen only in the cerebrospinal fluid injection group. Conclusion Dendritic cells are involved in the inner ear immune response to oti- tis media through all administration routes tested in this study. DCs are unique APCs and have been referred to as "pro- fessional" APCs. DCs have the ability to induce a primary immune response in the inner ear. Immune reaction can irre- versibly damage the structure and function of the inner ear. Our study suggests that early use of steroids may be effective in protecting the inner ear from immune damage.