Lamina propria plasma cells in inflammatory bowel disease: Intracellular detection of immunoglobulins using flow cytometry

被引:11
作者
Dorn, I
Schlenke, P
Mascher, B
Stange, EF
Seyfarth, M
机构
[1] Med Univ Lubeck, Inst Immunol & Transfus Med, D-23538 Lubeck, Germany
[2] Med Univ Lubeck, Dept Internal Med, D-23538 Lubeck, Germany
关键词
D O I
10.1078/0171-2985-00203
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
This is the first application of flow cytometry for the detection of lamina propria plasma cells and their intracellular immunoglobulins in patients with inflammatory bowel disease compared to healthy controls. The study has been focused on the distribution of IgA, IgG, IgM and the four IgG subclasses. Plasma cells were detected as high CD38 positive cells. For fixation and permeabilisation a single step reagent, Ortho Permeafix(R), was used. By flow cytometry, in patients with inflammatory bowel disease compared to healthy controls, a higher percentage of IgG(+) cells can be observed, in Crohn's disease also a higher percentage of IgM(+) cells. Regarding the IgG subclass distribution, patients with Crohns disease show an increase in IgG2(+) cells, patients with ulcerative colitis an increase in IgG1(+) and IgG3(+) cells. These results do agree with and expand the results of earlier immunohistochemical and functional studies, which are favoured today. For the determination of lymphocyte subset proportions and the detection of intracellular antigens, flow cytometry provides a useful alternative to well-established immunohistochemical methods. By analysing a larger number of cells, this method is more reproducible and less prone to inter-observer variations than immunohistochemistry, which needs the pre-selection of a mucosal area, the microscopic scoring of a limited number of cells and the circumvention of high background staining. The optimized flow cytometric protocol used in this study might be a promising tool for further investigations of various purposes.
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页码:546 / 557
页数:12
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