Schreiber S, MacDermott R P, Raedler A, Pinnau R, Bertovich M J, Nash G S
Gastrointestinal Section, University of Pennsylvania School of Medicine, Philadelphia.
Gastroenterology. 1991 Oct;101(4):1020-30. doi: 10.1016/0016-5085(91)90729-5.
Normal human lamina propria lymphocytes are in a heightened state of activation compared with peripheral blood with regard to cell-surface activation antigen expression (transferrin receptor, interleukin-2 receptor, 4F2) and the increased spontaneous secretion of immunoglobulins in vitro. This study evaluates the cell-surface expression of activation-associated antigens in different subpopulations of isolated colonic lamina propria mononuclear cells in inflammatory bowel disease. In pilot studies using three-color flow cytometry, autofluorescence was observed that was emitted by unstained lamina propria mononuclear cells, which interfered with both the sensitivity and the specificity of the analyses. Because a major portion of the intestinal lymphocyte populations of interest were autofluorescent, a method to remove autofluorescence signals was developed by designing a computer program for the subtraction of autofluorescence from the emissions of each individual cell. This technique increases both the sensitivity and specificity of flow-cytometric analyses of intestinal lamina propria mononuclear cells. Using fluorescence-activated cell-sorter analyses with subtraction of autofluorescence on a single-cell basis, increased expression of lymphocyte activation antigens (interleukin-2 receptor, transferrin receptor, 4F2) was found on the cell surface of isolated intestinal B cells, T cells, CD4+ T cells, and CD8+ T cells in both Crohn's disease and ulcerative colitis. Therefore, markedly increased intestinal lymphocyte activation is a major immunological alteration in inflammatory bowel disease and includes all lymphocyte subpopulations investigated in this study. In addition, 5-aminosalicylic acid, which is used for the treatment of intestinal inflammation in inflammatory bowel disease, inhibits the expression of cell-surface activation antigens on mitogen-activated peripheral blood lymphocytes in a dose-dependent manner. These observations suggest that lymphocyte activation may play an important role in underlying immune processes that lead to chronicity and perpetuation of inflammatory bowel disease and may implicate an additional mechanism for the therapeutic action of 5-aminosalicylic acid.
与外周血相比，正常人固有层淋巴细胞在细胞表面活化抗原表达（转铁蛋白受体、白细胞介素-2受体、4F2）以及体外免疫球蛋白自发分泌增加方面处于高度活化状态。本研究评估了炎症性肠病中分离的结肠固有层单核细胞不同亚群中活化相关抗原的细胞表面表达。在使用三色流式细胞术的初步研究中，观察到未染色的固有层单核细胞发出的自发荧光，这干扰了分析的敏感性和特异性。由于感兴趣的肠道淋巴细胞群体的很大一部分具有自发荧光，因此通过设计一个计算机程序来从每个细胞的发射信号中减去自发荧光，开发了一种去除自发荧光信号的方法。该技术提高了肠道固有层单核细胞流式细胞术分析的敏感性和特异性。使用基于单细胞减去自发荧光的荧光激活细胞分选分析，发现在克罗恩病和溃疡性结肠炎中，分离的肠道B细胞、T细胞、CD4+T细胞和CD8+T细胞的细胞表面淋巴细胞活化抗原（白细胞介素-2受体、转铁蛋白受体、4F2）表达增加。因此，肠道淋巴细胞活化明显增加是炎症性肠病的主要免疫学改变，包括本研究中调查的所有淋巴细胞亚群。此外，用于治疗炎症性肠病肠道炎症的5-氨基水杨酸以剂量依赖的方式抑制丝裂原激活的外周血淋巴细胞上细胞表面活化抗原的表达。这些观察结果表明，淋巴细胞活化可能在导致炎症性肠病慢性化和持续存在的潜在免疫过程中起重要作用，并且可能暗示了5-氨基水杨酸治疗作用的另一种机制。
