抗角蛋白抗体在活细胞反应部位的观察  

Observation on sites of antikeratin antibody in live keratinocyte

作　　者：武彩霞[1] 刘玉峰[1] 李承新[1] 夏汝山[1] 卢宁[1] 

机构地区：[1]第四军医大学西京医院皮肤科,陕西西安710033

出　　处：《第四军医大学学报》2003年第4期313-315,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金资助项目 (30 0 0 0 1 4 8)

摘　　要：目的 :观察抗角蛋白抗体在活细胞的反应部位 .方法 :以鼠抗人角蛋白的IgG单克隆抗体作用于培养中的Tca8113,以抗乙肝病毒表面抗原抗体做阴性对照 ,同时设置阳性对照和空白对照 .细胞固定、包埋后与FITC标记的羊抗鼠IgG结合 ,用荧光显微镜观察细胞的荧光着色 ;以胶体金标记的羊抗鼠IgG抗体染色 ,用透射电镜观察胶体金颗粒的部位 .结果 :抗角蛋白mAb作用的Tca8133细胞胞质呈亮绿色 ,着色较均匀 ,细胞核未见着色 .两种对照均未见着色 .抗角蛋白抗体作用的Tca8133细胞胞质的中间丝有胶体金颗粒 ,与阳性对照组类似 .阴性对照组及空白对照均未见胶体金颗粒 .结论 :抗角蛋白抗体可以进入活细胞 。AIM: To study the sites of anti keratin monoclonal antibody in live keratinocytes. METHODS: Human Tca cells were primarily cultured in vitro with antikeratin antibodies and some other Tca cells cultured with monoclonal antibodies to HBsAg served as the negative group. The positive group and the blank group were also established. All the cells were fixed and embedded and FITC labeled goat anti mouse IgG and ImmunoGold labeled anti mouse antibodies were added respectively. Then the cells were observed under the fluorescence microscopy and the electron microscope. RESULTS: Bright green fluorescence was seen in cytoplasm of Tca8113 cells cultured with anti keratin mAb. Stains of the cells were relatively homogeneous. No bright green fluorescence was seen in nucleuses of Tca8113 cells. And no bright green fluorescence was seen in Tca8113 cells cultured with HBsAb and malignant melanoma cultured with anti keratin mAb. ImmunoGold stained granules were seen in those cells cultured with antikeratin antibodies. The ImmunoGold stained granules appeared on the keratin intermediates. These phenomena were similar to what was seen in the positive group, while ImmunoGold stained granules were not seen in cells of the negative and blank groups. CONCLUSION: Antikeratin antibodies locate in the keratin intermediates of live keratinocyte cells.

关 键 词：抗角蛋白抗体 活细胞 反应部位 观察 

分 类 号：R392.11[医药卫生—免疫学]