Greenwood J H, Austin L L, Dobbins W O
Gastroenterology. 1983 Nov;85(5):1023-35.
Enriched populations of human intestinal and colonic intraepithelial lymphocytes were isolated separate from lamina propria lymphocytes. These populations, and peripheral blood mononuclear cells, were characterized for membrane features, lymphocyte subsets, and proliferative potential to the nonspecific mitogenic lectins phytohemagglutinin, concanavalin A, and pokeweed mitogen. All three populations were predominantly T lymphocytes as measured by sheep red blood cell rosettes and monoclonal antibodies. Peripheral blood mononuclear cells had a T4:T8 ratio of 2.15 while intraepithelial lymphocytes were enriched for T8+ cells with a T4:T8 ratio of 0.06-0.14. Peripheral blood mononuclear cells and lamina propria lymphocytes registered vigorous proliferative responses to phytohemagglutinin, concanavalin A, and pokeweed mitogen. Intraepithelial lymphocytes, in contrast, failed to respond to the lectins. Control experiments with peripheral blood mononuclear cells indicated that the isolation reagents and procedures had no adverse effect on lectin-stimulated proliferation or on T-cell subset proportions. Viability of the intraepithelial lymphocytes after separation (88%) and after 3 days in culture with lectin (77%-85%) was confirmed by trypan blue dye exclusion, by light microscopy, by electron microscopy, and by ability to affect pokeweed mitogen-induced immunoglobulin synthesis when added to autologous and heterologous peripheral blood mononuclear cells. The failure of intraepithelial lymphocytes to respond to lectins was not due to failure of lectin binding, macrophage depletion, or absence of T-cell growth factor in culture. The possibility that the T-cell subset proportions (T4:T8 ratio) affect the proliferative response of isolated intraepithelial lymphocytes is discussed.
从固有层淋巴细胞中分离出富集的人肠道及结肠上皮内淋巴细胞群体。对这些群体以及外周血单核细胞进行了膜特征、淋巴细胞亚群以及对非特异性促有丝分裂凝集素植物血凝素、刀豆球蛋白A和商陆有丝分裂原的增殖潜力的表征。通过绵羊红细胞玫瑰花结试验和单克隆抗体检测，所有这三个群体主要都是T淋巴细胞。外周血单核细胞的T4:T8比值为2.15，而上皮内淋巴细胞富含T8+细胞，T4:T8比值为0.06 - 0.14。外周血单核细胞和固有层淋巴细胞对植物血凝素、刀豆球蛋白A和商陆有丝分裂原表现出强烈的增殖反应。相比之下，上皮内淋巴细胞对这些凝集素无反应。对外周血单核细胞进行的对照实验表明，分离试剂和程序对凝集素刺激的增殖或T细胞亚群比例没有不利影响。通过台盼蓝拒染法、光学显微镜、电子显微镜以及将其添加到自体和异体外周血单核细胞中时影响商陆有丝分裂原诱导的免疫球蛋白合成的能力，证实了分离后（88%）以及与凝集素一起培养3天后（77% - 85%）上皮内淋巴细胞的活力。上皮内淋巴细胞对凝集素无反应并非由于凝集素结合失败、巨噬细胞耗竭或培养中缺乏T细胞生长因子。文中讨论了T细胞亚群比例（T4:T8比值）影响分离的上皮内淋巴细胞增殖反应的可能性。
