Lymphocyte gating of peripheral blood in patients with leukemic low-grade non-Hodgkin's lymphoma by multiparametric flow cytometry.
The standardized fluorescence intensity as expressed in molecules of equivalent soluble fluorescence (MESF) of lymphocytes from normal individuals and patients suffering from low-grade non-Hodgkin's-lymphomas was obtained comparing different staining patterns of CD45(FITC) and CD20(PerCP). After standardization of the flow cytometer using standardized fluorescent particles ('beads') significant differences could be obtained for hairy cell leukemia, chronic lymphocytic leukemia and immunocytomas in the peripheral blood. In contrast, centroblastic-centrocytic as well as centrocytic lymphomas showed no significant variations as compared to normal peripheral blood lymphocytes. According to these results, a new lymphocyte gating procedure was established by adding CD14(PE) and three-color measurement by CD45/CD14/CD20 staining of peripheral blood using erythrocyte lysis. The established gating procedure leads to a crucial discrimination and quantification of abnormal and normal lymphocytes per one measurement, whereas the 'leucogate' as defined by CD45/CD14 staining alone was insufficient for correct lymphocyte gate setting. In conclusion, the different staining of CD45 and CD20 in leukemic peripheral blood should be considered when fluorescence intensity or atypical peaks occurred in flow cytometric histograms suggesting for abnormal cell populations. In addition, it is possible to use this information to classify low-grade lymphomas.
Höffkes
H G
HG
Division of Hematology, Department of Medicine, University of Essen, Germany.
Schmidtke
G
G
Schmucker
U
U
Brittinger
G
G
D000328:Adult; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016130:Immunophenotyping; D008214:Lymphocytes / Q000276:immunology* / Q000473:pathology; D008228:Lymphoma, Non-Hodgkin / Q000097:blood* / Q000276:immunology / Q000473:pathology; D008297:Male; D008875:Middle Aged
D015703:Antigens, CD