Bartnik W, ReMine S G, Chiba M, Thayer W R, Shorter R G
Gastroenterology. 1980 May;78(5 Pt 1):976-85.
A technique is described, involving sequential treatment of the human colonic mucosa with EDTA in calcium-magnesium-free medium, and with collagenase, to isolate lymphoid cells enriched for intraepithelial (IEL) or lamina proprial lymphocytes (LPL). The IEL and LPL isolates also contained small numbers of eosinophils, mast cells, neutrophils, and macrophages. Plasma cells were present in the LPL but not in the IEL. The IEL isolates contained approximately equal proportions of T, B, and null cells. In contrast, the LPL suspensions contained 52% of T cells, 22% of B cells, and 26% of null cells. The most prevalent membrane immunoglobulin in the two colonic lymphoid cell suspensions was IgA (IEL--53%; LPL--71%). In colonic tissue sections, the percentages of immunoglobulin-containing cells as well as the proportions of cells containing IgA, both in the epithelial layer and the lamina propria, were similar to those found in the suspensions of lymphocytes stained for membrane immunoglobulin. These and other morphologic and characterization data support the contention that the two colonic lymphoid cell populations, obtained by the isolation procedures, were selectively enriched for intraepithelial or lamina proprial lymphocytes, respectively. Thus, this technique provides an important tool for further studies of the functional properties of the gut-associated lymphoid tissues.
本文描述了一种技术，该技术包括在无钙镁培养基中用乙二胺四乙酸（EDTA）对人结肠黏膜进行连续处理，然后用胶原酶处理，以分离富含上皮内淋巴细胞（IEL）或固有层淋巴细胞（LPL）的淋巴细胞。IEL和LPL分离物中还含有少量嗜酸性粒细胞、肥大细胞、中性粒细胞和巨噬细胞。浆细胞存在于LPL中，但不存在于IEL中。IEL分离物中T细胞、B细胞和裸细胞的比例大致相等。相比之下，LPL悬液中T细胞占52%，B细胞占22%，裸细胞占26%。两种结肠淋巴细胞悬液中最普遍的膜免疫球蛋白是IgA（IEL为53%；LPL为71%）。在结肠组织切片中，上皮层和固有层中含免疫球蛋白细胞的百分比以及含IgA细胞的比例，与膜免疫球蛋白染色的淋巴细胞悬液中的情况相似。这些以及其他形态学和特征数据支持这样的观点，即通过分离程序获得的两种结肠淋巴细胞群体分别选择性地富集了上皮内或固有层淋巴细胞。因此，该技术为进一步研究肠道相关淋巴组织的功能特性提供了重要工具。
