Non-vital polymorphonuclear leukocytes express myeloperoxidase on their surface.
The heme-containing enzyme myeloperoxidase (MPO) becomes expressed to the cell surface of non-vital polymorphonuclear leukocytes (PMNs) as evidenced by flow cytometry analysis and confocal fluorescence microscopy. While only a very small percentage of freshly isolated cells was able to bind the MPO antibody, PMN suspensions cultured for 36 h or longer time periods contained an increasing number of cells able to interact with these antibodies. Two distinct patterns of fluorescence for the MPO antibodies were observed. Antibodies were localised either in surface patches or distributed over the whole cell body. The latter type dominated in cell samples cultured for more than three days, while the first type was predominantly found in samples cultured for lower time periods. We observed also two peaks for fluorescence distribution by flow cytometry after addition of MPO antibodies to PMNs. Myeloperoxidase was localised at phosphatidylserine epitopes at the surface of non-vital PMNs as evidenced by coincubation with fluorescent MPO antibodies and FITC-labelled annexin V. Myeloperoxidase bound to the outer surface of PMNs uses hydrogen peroxide as a substrate as shown by appearance of an intense chemiluminescence using the impermeable luminescent protein Pholasin. Thus, myeloperoxidase becomes expressed to the surface of non-vital polymorphonuclear leukocytes colocalised with phosphatidylserine that may indicate a role of myeloperoxidase in apoptosis of PMNs.
Flemmig
Jorg
J
Institute for Medical Physics and Biophysics, Medical Faculty, University of Leipzig, Germany.
Lessig
Jacqueline
J
Reibetanz
Uta
U
Dautel
Philipp
P
Arnhold
Jurgen
J
D017304:Annexin A5 / Q000378:metabolism; D000906:Antibodies / Q000276:immunology; D017209:Apoptosis; D002462:Cell Membrane / Q000201:enzymology*; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D018613:Microscopy, Confocal; D009504:Neutrophils / Q000166:cytology* / Q000201:enzymology* / Q000276:immunology; D009195:Peroxidase / Q000378:metabolism*; D011485:Protein Binding; D013379:Substrate Specificity; D013997:Time Factors
D017304:Annexin A5; D000906:Antibodies; D006861:Hydrogen Peroxide; D009195:Peroxidase