Flow cytometric analysis of intracellular myeloperoxidase distinguishes lymphocytes, monocytes and granulocytes.
A study was undertaken to evaluate the ability of flow cytometric analysis of intracellular myeloperoxidase (MPO) in differentiating populations of lymphocytes (L), monocytes (M) and granulocytes (G), by means of lysed whole blood method. Anticoagulated blood from 23 normal individuals was lysed with FACS lysing solution and permeabilized with FACS permeabilizing solution before subjected to direct immunofluorescence staining. The geometric means of the fluorescence intensity were measured using FACSCalibur flow cytometer (Becton Dickinson). Populations of L, M and G were gated based on their light scatter characteristics and expression of CD14 and CD45. Then, the fluorescence intensity of MPO expression was studied in these individual cell populations. The results showed that fluorescence intensity of MPO was the strongest in G and weakest in L, whereas M showed intermediate fluorescence intensity. Our findings reveal that discrimination of these three cell types is achievable based upon the sole expression of intracellular MPO.
Tay
S P
SP
Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
Cheong
S K
SK
Hamidah
N H
NH
Ainoon
O
O
D000954:Antigens, Surface / Q000032:analysis; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D019085:Fluorescent Antibody Technique, Direct; D006098:Granulocytes / Q000201:enzymology*; D006461:Hemolysis; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology*; D008297:Male; D009000:Monocytes / Q000201:enzymology*; D009195:Peroxidase / Q000276:immunology / Q000378:metabolism*
D000954:Antigens, Surface; D009195:Peroxidase