Mature eosinophils stimulated to develop in human-cord blood mononuclear cell cultures supplemented with recombinant human interleukin-5. II. Vesicular transport of specific granule matrix peroxidase, a mechanism for effecting piecemeal degranulation.
The mechanism of piecemeal degranulation by human eosinophils was investigated. Mature eosinophils that developed in rhIL-5-containing conditioned media from cultured human cord blood mononuclear cells were prepared for ultrastructural studies using a combined technique to image eosinophil peroxidase by cytochemistry in the same sections on which postembedding immunogold was used to demonstrate Charcot-Leyden crystal protein. Vesicular transport of eosinophil peroxidase from the specific granule matrix compartment to the cell surface was associated with piecemeal degranulation. This process involved budding of eosinophil peroxidase-loaded vesicles and tubules from specific granules. Some eosinophil peroxidase that was released from eosinophils remained bound to the cell surface; some was free among the cultured cells. Macrophages and basophils bound the released eosinophil peroxidase to their plasma membranes, internalized it in endocytotic vesicles, and stored it in their respective phagolysosomes and secretory granules. Charcot-Leyden crystal protein was diffusely present in the nucleus and cytoplasm of IL-5-stimulated mature eosinophils. Extensive amounts were generally present in granule-poor and subplasma membrane areas of the cytoplasm in contrast to eosinophil peroxidase, which was secreted and bound to the external surface of eosinophil plasma membranes. These studies establish vesicular transport as a mechanism for emptying the specific eosinophil granule matrix compartment during IL-5-associated piecemeal degranulation.
Dvorak
A M
AM
Department of Pathology, Beth Israel Hospital, Boston, MA 02215.
Ackerman
S J
SJ
Furitsu
T
T
Estrella
P
P
Letourneau
L
L
Ishizaka
T
T
D001773:Blood Cells / Q000166:cytology*; D002478:Cells, Cultured; D016922:Cellular Senescence; D003594:Cytoplasmic Granules / Q000378:metabolism; D004804:Eosinophils / Q000166:cytology* / Q000378:metabolism; D005312:Fetal Blood; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D015848:Interleukin-5 / Q000494:pharmacology*; D008245:Lysophospholipase; D009000:Monocytes / Q000166:cytology*; D010544:Peroxidases / Q000378:metabolism; D011994:Recombinant Proteins; D014018:Tissue Distribution
D006023:Glycoproteins; D015848:Interleukin-5; D011994:Recombinant Proteins; D010544:Peroxidases; D008245:Lysophospholipase; C011711:lysolecithin acylhydrolase