| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
American Journal of Pathology, Vol 138, 69-82, Copyright © 1991 by American Society for Investigative Pathology
REGULAR ARTICLES |
AM Dvorak, T Furitsu, L Letourneau, T Ishizaka and SJ Ackerman
Department of Pathology, Beth Israel Hospital.
Human cord blood mononuclear cells were cultured for 35 days in media containing recombinant human interleukin 5 (rhIL-5) supplemented with a fraction of the culture supernatant of phytohemagglutinin (PHA)- stimulated human T lymphocytes from which interleukin 2 (IL-2) was eliminated. Cultured cells were studied by electron microscopy and an immunogold procedure to detect subcellular site(s) of Charcot-Leyden crystal (CLC) protein. The majority of cells (greater than 70%) developing in this system were mature eosinophils, with descending frequency of other cells, including macrophages, mature basophils, eosinophilic myelocytes, and mature neutrophils. Mature eosinophils were characterized by increased numbers of primary granules, small granules, tubulovesicular structures, and decreased secondary granules. These eosinophils showed extensive piecemeal degranulation (PMD) characterized by partially empty and empty secondary granule chambers in the cytoplasm. Small, smooth vesicles were evident within empty granule chambers as well as adjacent to them. Eosinophils formed close associations with phagocytic macrophages that contained both standard- shaped and irregularly shaped CLC within phagolysosomes. Subcellular sites of CLC protein were demonstrated by immunogold in eosinophils and macrophages arising in these cultures. Charcot-Leyden crystal protein was present in the nuclear matrix and extraorganellar cytoplasm of eosinophils. Primary granules and some cytoplasmic vesicles were labeled for CLC protein, but full and empty secondary granules and the extensive network of tubulovesicles were not. Charcot-Leyden crystals were absent from eosinophils, nor were they present in the extracellular space. Charcot-Leyden crystals were absent from eosinophils, nor were they present in the extracellular space. Charcot- Leyden crystals within phagosomes of macrophages were labeled by the immunogold procedure for CLC protein. These results demonstrate that rhIL-5-supplemented, PHA-stimulated, T-cell-conditioned media induced the development of mature human eosinophils from cord blood cells. These eosinophils underwent PMD of secondary granule contents. Immunogold analysis showed eosinophil CLC protein in the cytoplasm, nucleus, and primary granules of eosinophils. Macrophages also were present in these cultures. They contained CLC protein-containing crystals in their phagosomes, suggesting active sequestration of eosinophil CLC protein by macrophages in vitro.
This article has been cited by other articles:
 |
|
 |
|
  L. A. Spencer, C. T. Szela, S. A. C. Perez, C. L. Kirchhoffer, J. S. Neves, A. L. Radke, and P. F. Weller Human eosinophils constitutively express multiple Th1, Th2, and immunoregulatory cytokines that are secreted rapidly and differentially J. Leukoc. Biol., January 1, 2009; 85(1): 117 - 123. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. C. N. Melo, L. A. Spencer, A. M. Dvorak, and P. F. Weller Mechanisms of eosinophil secretion: large vesiculotubular carriers mediate transport and release of granule-derived cytokines and other proteins J. Leukoc. Biol., February 1, 2008; 83(2): 229 - 236. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Moqbel and J. J. Coughlin Differential Secretion of Cytokines Sci. Signal., June 6, 2006; 2006(338): pe26 - pe26. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. A. Spencer, R. C. N. Melo, S. A. C. Perez, S. P. Bafford, A. M. Dvorak, and P. F. Weller Cytokine receptor-mediated trafficking of preformed IL-4 in eosinophils identifies an innate immune mechanism of cytokine secretion PNAS, February 28, 2006; 103(9): 3333 - 3338. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Clark, L. Simson, N. Newcombe, A. M. L. Koskinen, J. Mattes, N. A. Lee, J. J. Lee, L. A. Dent, K. I. Matthaei, and P. S. Foster Eosinophil degranulation in the allergic lung of mice primarily occurs in the airway lumen J. Leukoc. Biol., June 1, 2004; 75(6): 1001 - 1009. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Mahmudi-Azer, G. P. Downey, and R. Moqbel Translocation of the tetraspanin CD63 in association with human eosinophil mediator release Blood, May 13, 2002; 99(11): 4039 - 4047. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. L. Denzler, M. T. Borchers, J. R. Crosby, G. Cieslewicz, E. M. Hines, J. P. Justice, S. A. Cormier, K. A. Lindenberger, W. Song, W. Wu, et al. Extensive Eosinophil Degranulation and Peroxidase-Mediated Oxidation of Airway Proteins Do Not Occur in a Mouse Ovalbumin-Challenge Model of Pulmonary Inflammation J. Immunol., August 1, 2001; 167(3): 1672 - 1682. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Feng, R. Flaumenhaft, C. Bandeira–Melo, P. F. Weller, and A. M. Dvorak Ultrastructural Localization of Vesicle-associated Membrane Protein(s) to Specialized Membrane Structures in Human Pericytes, Vascular Smooth Muscle Cells, Endothelial Cells, Neutrophils, and Eosinophils J. Histochem. Cytochem., March 1, 2001; 49(3): 293 - 304. [Abstract] [Full Text]
|
|
|
|
 |
|
 J. S. ERJEFÄLT and C. G. A. PERSSON New Aspects of Degranulation and Fates of Airway Mucosal Eosinophils Am. J. Respir. Crit. Care Med., June 1, 2000; 161(6): 2074 - 2085. [Full Text]
|
|
|
|
|
|
J. S. ERJEFÄLT, L. GREIFF, M. ANDERSSON, E. MATSSON, H. PETERSEN, M. LINDEN, T. ANSARI, P. K. JEFFERY, and C. G. A. PERSSON Allergen-induced Eosinophil Cytolysis Is a Primary Mechanism for Granule Protein Release in Human Upper Airways Am. J. Respir. Crit. Care Med., July 1, 1999; 160(1): 304 - 312. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. A. Giembycz and M. A. Lindsay Pharmacology of the Eosinophil Pharmacol. Rev., June 1, 1999; 51(2): 213 - 340. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. B. Huffnagle, M. B. Boyd, N. E. Street, and M. F. Lipscomb IL-5 Is Required for Eosinophil Recruitment, Crystal Deposition, and Mononuclear Cell Recruitment During a Pulmonary Cryptococcus neoformans Infection in Genetically Susceptible Mice (C57BL/6) J. Immunol., March 1, 1998; 160(5): 2393 - 2400. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Scepek and M. Lindau Exocytotic Competence and Intergranular Fusion in Cord Blood-Derived Eosinophils During Differentiation Blood, January 15, 1997; 89(2): 510 - 517. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M Lindau, O Nusse, J Bennett, and O Cromwell The membrane fusion events in degranulating guinea pig eosinophils J. Cell Sci., January 1, 1993; 104(1): 203 - 210. [Abstract] [PDF]
|
|
|
|
 |
|
 A. Y. Huang, A. M. Castle, B. T. Hinton, and J. D. Castle Resting (Basal) Secretion of Proteins Is Provided by the Minor Regulated and Constitutive-like Pathways and Not Granule Exocytosis in Parotid Acinar Cells J. Biol. Chem., June 15, 2001; 276(25): 22296 - 22306. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
