小鼠面神经损伤急性期T细胞CD69表达的研究

The expression of CD69 on T cells of the mouse with facial nerve transection at acute stage

目的应用流式细胞技术探讨面神经损伤急性期T细胞功能状态及其病理生理意义,为揭示外伤性面瘫的神经免疫机制提供理论依据。方法制备面神经轴切损伤模型小鼠,分别于损伤后3天、2周分离小鼠肠系膜淋巴结(mesenteric lymph node,MLN)、术侧颈部引流淋巴结(cervical draininglymph node,CDLN)细胞,进行双色免疫荧光标记,以流式细胞术分析T细胞上CD69分子的表达情况。结果手术后3天,面神经损伤组颈部引流淋巴结T细胞CD69表达的百分率与相应手术对照组相比较差异有显著性(P=0.0457);而神经损伤组、手术对照组肠系膜淋巴结T细胞CD69表达的百分率与正常小鼠相比较差异无显著性(P值分别为0.2817、0.2724)。面神经轴切损伤后2周,神经损伤组CDLN的T细胞CD69表达的百分率仍然维持在较高水平,神经损伤组MLN的T细胞CD69表达的百分率出现上调,与相应对照组及术后3天时相比较差异均有显著性(P值分别为0.0082、0.0133)。结论面神经轴切损伤3天、2周时,颈部引流淋巴结存在T细胞活化并上调;在2周时,肠系膜淋巴结也出现低水平的T细胞活化。提示面神经损伤急性期,伴随着一个局部免疫应答向全身免疫应答转化的过程,在一定程度上有利于机体协调控制免疫应答的规模与方向。

Objective To investigate the T cell activation of the facial nerve axotomized mouse with methods of flow cytometry for the latent neuroimmunologic mechanism of traumatic facial paralysis. Methods The murine model with facial nerve transaction was established, then the lymphocytes separated from the cervical drainage lymphoid nodes （CDLN） and mesenteric lymph nodes （MLN） of BABL/c mouse at specific time were collected and estimated the expression rate of CD69 on T cells by flow cytometry following two-color immunofluorescent staining. Results Three days post the operation （DPO3）, the expression rate of CD69 on the T cells from the cervical drainage lymphoid nodes of the groups with facial nerve transection had significant difference with that of the MLN from group with facial nerve transected （P = 0.0457）, whereas the expression rates of CD69 on the T cells from the mesenteric lymph nodes of the groups with facial nerve transection and the groups with control operation both had no significant difference compared with the expression rate of the blank group （P = 0.2817 and 0.2724, respectively）. Two weeks post the operation （DPO14）, the expression rate of CD69 on the T cells from the cervical drainage lymphoid nodes of the groups with facial nerve transection was still at a higher level, it had a significant difference with that of the sham-operation group （P = 0.0007 ） ; also the expression rates of CD69 on the T cells from the mesenteric lymph nodes of the groups with facial nerve transection was upregulated, it had a significant difference compared with the expression rates both of the groups with sham-operation DPO14 and with facial nerve transaction DPO3 （P = 0.0082 and 0.0133, respectively ）. Conclusion There was T cell activation and up-regulation of the cervical drainage lymphoid nodes from the mouse model with facial nerve transaction, even there was T cell activation of the mesenteric lymph nodes at 2 weeks post operation. It suggested a alteration of immune response from local immunity to general immunity while facial nerve trauma at the acute stage, which probably profited individuals coordinating and controlling the scales and orientation of the neuroimmune response during the pathogenesis and progression of facial nerve trauma.