Comparative properties of the Charcot-Leyden crystal protein and the major basic protein from human eosinophils.

Gleich, Gerald J.; Loegering, David A.; Mann, Kenneth G.; Maldonado, Jorge E.

doi:10.1172/jci108319

Cited by 215 publications

(74 citation statements)

References 14 publications

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“…Lymphocytes and neutrophils were obtained from the blood of normal volunteers as described (22) and were >95% pure. Eosinophils were obtained from guinea pigs by the methods of Gleich and Loegering (23) and Pincus (24); these cells were >95% pure. Each of these cell populations was cultured in microtiter wells in RPMI-1640 medium without serum under similar conditions to those used for alveolar macrophages.…”

Section: Methodsmentioning

confidence: 99%

Production of fibronectin by the human alveolar macrophage: mechanism for the recruitment of fibroblasts to sites of tissue injury in interstitial lung diseases.

Rennard¹,

Hunninghake²,

Bitterman³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

289

110

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Because cells of the mononuclear phagocyte system are known to produce fibronectin and because alveolar macrophages are activated in many interstitial lung diseases, the present study was designed to evaluate a-role for the alveolar macrophage as a source ofthe increased levels offibronectin found in the lower respiratory tract in interstitial lung. diseases and to determine ifsuch fibronectin might contribute to the development of the fibrosis found in these disorders by beinga chemoattractant for human lung fibroblasts. Production of fibronectin by human alveolar. macrophages obtained by bronchoalveolar lavage and maintained in short-term culture in serum-free conditions was demonstrated; de novo synthesis was confirmed by the incorporation of ['4C]proline. This fibronectin had a monomer molecular weight of 220,000 and was antigenically similar to plasma fibronectin. Macrophages from patients with idiopathic pulmonary fibrosis produced fibronectin at a rate 20 times higher than did normal macrophages; macrophages from patients with pulmonary sarcoidosis produced fibronectin at 10 times the normal rate. Macrophages from 6 of 10 patients with various other interstitial disorders produced fibronectin at rates greater than the rate of highest normal control. Human alveolar macrophage fibronectin was chemotactic for human lung fibroblasts, suggesting a functional role for this fibronectin in the derangement of the alveolar structures that is characteristic of these disorders.Fibronectin, a large glycoprotein found in plasma and tissues, is thought to mediate cell-matrix interactions (1-4) through its ability to bind to cells (5, 6), collagen (7,8), and other connective tissue components and to attract cells by virtue of its chemotactic properties (9, 10). Several cell types are known to produce this macromolecule, including cells of the mononuclear phagocyte system (11,12). Together, these observations lead to the hypothesis that mononuclear phagocytes at sites oftissue injury produce increased amounts of fibronectin which, in turn, recruits fibroblasts to aid in the repair process.The lung is a common site of tissue injury and is one of the few sites where human tissue mononuclear phagocytes are readily available for study. In addition, recent studies have demonstrated increased levels of fibronectin in the lower respiratory tract of patients with interstitial lung diseases (13). In this context, the present study was designed to answer several questions, including (i) do human alveolar macrophages produce fibronectin? (ii) do alveolar macrophages from patients with chronic tissue injury produce increased amounts of fibronectin? and (iii) is the fibronectin produced by the human alveolar macrophage chemotactic for human lung fibroblasts? MATERIALS AND METHODSStudy Population. The study population consisted of 48 patients with interstitial lung disease and 7 normal volunteers. Patients were selected from consecutive patients evaluated for interstitial lung disease at the National Institutes of Health and f...

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Section: Methodsmentioning

confidence: 99%

Production of fibronectin by the human alveolar macrophage: mechanism for the recruitment of fibroblasts to sites of tissue injury in interstitial lung diseases.

Rennard¹,

Hunninghake²,

Bitterman³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

289

110

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“…Native MBP was purified from eosinophils of patients with marked peripheral blood eosinophilia as described in detail elsewhere (14). Sephadex G-50 fractions containing native MBP, in 0.025 M sodium acetate column buffer, pH 4.3, were pooled and stored in aliquots at -70°C.…”

Section: Isolation Of Rat Peritoneal Mast Cellsmentioning

confidence: 99%

Activation of basophil and mast cell histamine release by eosinophil granule major basic protein.

O’Donnell

Ackerman

Gleich

et al. 1983

The Journal of Experimental Medicine

Self Cite

313

107

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Major basic protein (MBP) is a primary constituent of eosinophil granules. In this report, we demonstrate that MBP from human eosinophil granules initiates a nonlytic histamine release from human leukocytes. A direct effect of MBP on basophils was confirmed using purified human basophils. The kinetics of release were similar to those reported for poly-L-arginine, although MBP was less potent than poly-L-arginine of similar molecular weight. Reduction and alkylation of MBP diminished both the potency and efficacy of the molecule. Native MBP also stimulated histamine secretion from purified rat peritoneal mast cells in a manner characteristic of other polycations. These results emphasize the bidirectional nature of the basophil/mast cell-eosinophil regulatory axis.

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“…CLC protein is the sole protein constituent of CLCs purified from human eosinophils; it expresses lysophospholipase activity, forms these characteristic crystals, and comprises an estimated 7-10% of total cellular protein in the eosinophil (9)(10)(11)(12). CLCs and the CLC protein, once considered unique to the eosinophil lineage, have also been found in human basophils (2,13,14).…”

Section: Introductionmentioning

confidence: 99%

Charcot-Leyden crystal protein distribution in basophils and its absence in mast cells that differentiate from human umbilical cord blood precursor cells cultured in murine fibroblast culture supernatants or in recombinant human c-kit ligand.

Dvorak

Letourneau

Albee

et al. 1994

J Histochem Cytochem.

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Suspension cultures of human umbilical cord blood mononuclear cells supplemented with c-kit ligand-containing additives give rise to a mixture of cells belonging to several lineages. Among those that differentiate in quantity are mature basophils, immature mast cells, and neutrophilic myelocytes. We used an ultrastructural immunogold method to detect the Charcot-Leyden crystal (CLC) protein, an eosinophiland basophil-specific protein, to study cells that were obtained at sequential times from 3 to 14 weeks in culture. Basophils (and eosinophils, which were present in smaller numbers) labeled for the CLC protein; mast cells did not. The labeled basophil subcellular sites included formed intragranular, cytoplasmic and nuclear CLCs, cytoplasmic particle-filled and homogeneously dense granules, cytoplasm, nucleus, plasma membrane, and cytoplasmic and IntroductionHuman basophils and mast cells can now be reliably induced to develop in vitro from their agranular precursors which are present in cord blood (1-6). Initially, these studies were done with supplements of various cell-conditioned media now known to contain a basophil growth factor, interleukin-3 (IL3) (l), or a mast cell growth factor, the c-kit ligand [also called stem cell factor (SCF)] (5,6). Some of these studies were also done with cord blood cells cocultured with mouse 3T3 fibroblasts (3,4 repeated with these reagents. In all of these cultures, ultrastructural criteria were used to assign immature and mature cells to the appropriate lineages (1-5) (AM Dvorak et al., unpublished data).Charcot-Leyden crystals (CLCs) are hexagonal, bipyramidal structures originally described in eosinophil-rich disorders (7,8). CLC protein is the sole protein constituent of CLCs purified from human eosinophils; it expresses lysophospholipase activity, forms these characteristic crystals, and comprises an estimated 7-10% of total cellular protein in the eosinophil (9-12). CLCs and the CLC protein, once considered unique to the eosinophil lineage, have also been found in human basophils (2,13,14). Using an ultrastructural immunogold method, we have defined the subcellular locations of the CLC protein in quiescent peripheral blood eosinophils (15) and basophils (16) (17-19). Although the cellular function(s) of this lysophospholipase is still obscure, the presence of such large quantities of this enzyme in eosinophils and basophils suggests an important role in normal or pathological events associated with these lineages. The recent cloning of the gene for the CLC protein (20) and expression of the protein product (12) should aid future studies of the biological properties of this unique basophil and eosinophil constituent.Human mast cells, in contrast to basophils, do not stain for the CLC protein by light microscopy (21). Since we noted an admixture of lineages in suspension cultures of cord blood supplemented with c-kit ligand-containing additives, we examined these cells for CLC protein using an immunogold method to detect this protein.We found that mature basophils that ...

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Comparative properties of the Charcot-Leyden crystal protein and the major basic protein from human eosinophils.

Cited by 215 publications

References 14 publications

Production of fibronectin by the human alveolar macrophage: mechanism for the recruitment of fibroblasts to sites of tissue injury in interstitial lung diseases.

Production of fibronectin by the human alveolar macrophage: mechanism for the recruitment of fibroblasts to sites of tissue injury in interstitial lung diseases.

Activation of basophil and mast cell histamine release by eosinophil granule major basic protein.

Charcot-Leyden crystal protein distribution in basophils and its absence in mast cells that differentiate from human umbilical cord blood precursor cells cultured in murine fibroblast culture supernatants or in recombinant human c-kit ligand.

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