Qiu Zhijuan, Sheridan Brian S
Center for Infectious Diseases, Department of Molecular Genetics and Microbiology, Stony Brook University.
Center for Infectious Diseases, Department of Molecular Genetics and Microbiology, Stony Brook University;
J Vis Exp. 2018 Feb 28(132):57281. doi: 10.3791/57281.
The intestinal immune system plays an essential role in maintaining the barrier function of the gastrointestinal tract by generating tolerant responses to dietary antigens and commensal bacteria while mounting effective immune responses to enteropathogenic microbes. In addition, it has become clear that local intestinal immunity has a profound impact on distant and systemic immunity. Therefore, it is important to study how an intestinal immune response is induced and what the immunologic outcome of the response is. Here, a detailed protocol is described for the isolation of lymphocytes from small intestine inductive sites like the gut-associated lymphoid tissue Peyer's patches and the draining mesenteric lymph nodes and effector sites like the lamina propria and the intestinal epithelium. This technique ensures isolation of a large numbers of lymphocytes from small intestinal tissues with optimal purity and viability and minimal cross compartmental contamination within acceptable time constraints. The technical capability to isolate lymphocytes and other immune cells from intestinal tissues enables the understanding of immune responses to gastrointestinal infections, cancers, and inflammatory diseases.
肠道免疫系统通过对饮食抗原和共生细菌产生耐受性反应,同时对肠道致病微生物发起有效的免疫反应,在维持胃肠道屏障功能方面发挥着至关重要的作用。此外,很明显局部肠道免疫对远处和全身免疫有深远影响。因此,研究肠道免疫反应是如何诱导的以及该反应的免疫结果是什么很重要。在此,描述了一种详细的方案,用于从小肠诱导部位(如肠道相关淋巴组织派尔集合淋巴结和引流肠系膜淋巴结)以及效应部位(如固有层和肠上皮)分离淋巴细胞。该技术可确保在可接受的时间限制内,从小肠组织中分离出大量纯度最佳、活力最佳且跨区污染最小的淋巴细胞。从肠道组织中分离淋巴细胞和其他免疫细胞的技术能力有助于理解对胃肠道感染、癌症和炎症性疾病的免疫反应。