Ectosomes released by human neutrophils are specialized functional units.
Here we show that human polymorphonuclear leukocytes (PMN) release ectosomes independently of complement attack during their activation both in vitro and at the site of inflammation in vivo. Patterns of biotinylated proteins on the surface of PMN and on PMN-derived ectosomes indicated a specific sorting of cell surface proteins into and out of ectosomes. Ectosomes expressed clusters of complement receptor 1 (CR1), which allowed them to bind efficiently to opsonized bacteria. Myeloperoxidase and human leukocyte elastase, both stored within the azurophilic granules of PMN, were found to colocalize on ectosomes with CR1. Furthermore, myeloperoxidase colocalized with human leukocyte elastase. In contrast, not present on CR1-expressing ectosomes were CD63, a selective marker for the azurophilic granules, and CD14, which is located within the same granules and the secretory vesicles as CR1. Of the other complement regulatory proteins expressed by PMN, only CD59 colocalized with CR1, while CD55 and CD46 were almost absent. Ectosomes released by activated PMN at the site of inflammation may function as a well organized element (ecto-organelle), designed to focus antimicrobial activity onto opsonized surfaces.
Hess
C
C
Department of Research, University Hospital Basel, Switzerland. hessch@ubaclu.unibas.ch
Sadallah
S
S
Hefti
A
A
Landmann
R
R
Schifferli
J A
JA
D000818:Animals; D019921:Biotinylation; D001768:Blister / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology / Q000276:immunology; D002469:Cell Separation; D018699:Coated Vesicles / Q000276:immunology* / Q000378:metabolism* / Q000648:ultrastructure; D004912:Erythrocytes / Q000276:immunology; D006461:Hemolysis; D006801:Humans; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism* / Q000382:microbiology / Q000648:ultrastructure; D011506:Proteins / Q000378:metabolism; D011817:Rabbits; D017463:Receptors, Complement 3b / Q000378:metabolism / Q000648:ultrastructure
D011506:Proteins; D017463:Receptors, Complement 3b