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Updated: Aug 7, 2026
Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes
Published on: May 8, 2016
Lamina propria plasma cells in inflammatory bowel disease: intracellular detection of immunoglobulins using flow
Isabel Dorn1, Peter Schlenke, Beate Mascher
1Institute of Immunology and Transfusion Medicine, University of Luebeck School of Medicine, Luebeck, Germany. dorn@immu.mu-luebeck.de
Insights
Flow cytometry effectively detects plasma cells and immunoglobulins in inflammatory bowel disease (IBD). This method reveals distinct immunoglobulin patterns in Crohn's disease and ulcerative colitis, offering a reproducible alternative to immunohistochemistry.
Area of Science:
- Immunology
- Gastroenterology
- Cell Biology
Background:
- Inflammatory bowel disease (IBD) involves complex immune dysregulation.
- Understanding immunoglobulin distribution in the lamina propria is crucial for IBD pathogenesis.
- Current methods like immunohistochemistry have limitations in reproducibility and cell analysis.
Purpose of the Study:
- To apply flow cytometry for quantifying lamina propria plasma cells and their intracellular immunoglobulins in IBD patients.
- To analyze the distribution of IgA, IgG, IgM, and IgG subclasses in IBD.
- To establish flow cytometry as a reliable alternative to immunohistochemistry for IBD research.
Main Methods:
- Flow cytometry was used to detect plasma cells (CD38+).
- Single-step fixation and permeabilization (Ortho Permeafix) were employed.
- Intracellular immunoglobulin staining (IgA, IgG, IgM, IgG subclasses) was performed.
Main Results:
- IBD patients showed higher percentages of IgG+ cells compared to controls.
- Crohn's disease patients had increased IgM+ cells.
- Crohn's disease exhibited elevated IgG2+ cells, while ulcerative colitis showed increased IgG1+ and IgG3+ cells.
Conclusions:
- Flow cytometry is a valuable tool for analyzing plasma cells and immunoglobulins in IBD.
- The study identified specific immunoglobulin subclass alterations in Crohn's disease and ulcerative colitis.
- Flow cytometry offers a more reproducible and quantitative approach than traditional immunohistochemistry for IBD research.
Abstract:
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, 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 Crohn's 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 interobserver 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.
