论文部分内容阅读
目的：建立一种提取小肠上皮内淋巴细胞的改良方法，对其功能进行初步研究。方法：采用机械分离法和Ｐｅｒｃｏｌｌ非连续密度梯度离心法提取小肠上皮内淋巴细胞，经间接免疫荧光染色后用流式细胞仪检测表型。应用乳酸脱氢酶法检测其自然杀伤功能。结果：应用本方法可获得小鼠小肠上皮内淋巴细胞２～４×１０６ 个／只鼠，活力测定＞ ９５％ 。抗原表型中７３．８％ 以上表达ＣＤ８ 分子，ＴＣＲγδ为４３．８％ 。而且自然杀伤活性明显高于脾中淋巴细胞。结论：应用机械分离法和Ｐｅｒｃｏｌｌ非连续密度梯度离心法提取小肠上皮内淋巴细胞的方法有效，ＩＥＬ具有自然杀伤活性
OBJECTIVE: To establish an improved method for extracting intra-intestinal epithelial lymphocytes and to study its function. Methods: The intestinal epithelial lymphocytes were isolated by mechanical separation and Percoll non-continuous density gradient centrifugation. Phenotypes were detected by flow cytometry after indirect immunofluorescence staining. Application of lactate dehydrogenase method to detect its natural killer function. Results: 2 ~ 4 × 106 mouse intestinal epithelial lymphocytes were obtained by this method and the viability was> 95%. Over 73.8% of the antigenic epitopes expressed CD8 molecules with a TCRγδ of 43.8%. And natural killer activity was significantly higher than the spleen lymphocytes. Conclusion: The method of mechanical separation and Percoll discontinuous density gradient centrifugation to extract intestinal intraepithelial lymphocytes is effective. IEL has natural killer activity