Lamina propria plasma cells in inflammatory bowel disease: intracellular detection of immunoglobulins using flow cytometry
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Lamina propria plasma cells in inflammatory bowel disease: intracellular detection of immunoglobulins using flow cytometry. / Dorn, Isabel; Schlenke, Peter; Mascher, Beate; Stange, Eduard Friedrich; Seyfarth, Michael.
in: IMMUNOBIOLOGY, Jahrgang 206, Nr. 5, 12.2002, S. 546-57.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Lamina propria plasma cells in inflammatory bowel disease: intracellular detection of immunoglobulins using flow cytometry
AU - Dorn, Isabel
AU - Schlenke, Peter
AU - Mascher, Beate
AU - Stange, Eduard Friedrich
AU - Seyfarth, Michael
PY - 2002/12
Y1 - 2002/12
N2 - 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.
AB - 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.
KW - ADP-ribosyl Cyclase/metabolism
KW - ADP-ribosyl Cyclase 1
KW - Adult
KW - Antigens, CD/metabolism
KW - Colon/immunology
KW - Female
KW - Flow Cytometry/methods
KW - Humans
KW - Immunoglobulins/analysis
KW - Inflammatory Bowel Diseases/immunology
KW - Intestinal Mucosa/immunology
KW - Intracellular Fluid/chemistry
KW - Male
KW - Membrane Glycoproteins
KW - Middle Aged
KW - Plasma Cells/chemistry
U2 - 10.1078/0171-2985-00203
DO - 10.1078/0171-2985-00203
M3 - SCORING: Journal article
C2 - 12607730
VL - 206
SP - 546
EP - 557
JO - IMMUNOBIOLOGY
JF - IMMUNOBIOLOGY
SN - 0171-2985
IS - 5
ER -