Cough responses to inhaled irritants are enhanced by eosinophil major basic protein in awake mice

A distinct association between airway eosinophilia and chronic cough is well documented. Eosinophil granule-derived cationic proteins, such as major basic protein (MBP), have been shown to activate and enhance the excitability of bronchopulmonary C-fiber sensory nerves, which may then lead to an increase in cough sensitivity. This study was carried out to determine whether cough responses to inhaled irritant gases were altered by delivery of MBP into the airways. An awake mouse moved freely in a recording chamber that was ventilated with a constant flow of air or irritant gas mixture. Cough responses to separate inhalation challenges of sulfur dioxide (SO2; 300 and 600 ppm) and ammonia (NH3; 0.1 and 0.2%), each for 5-min duration, were measured daily for 3 days before and for up to 8 days after MBP (10–20 µg) instillation into the trachea. During control, inhalations of SO2 and NH3 consistently elicited cough responses in a dose-dependent manner. After MBP treatment, cough responses to both SO2 and NH3 increased significantly and progressively and reached peaks 2–3 days after the treatment before returning to control level in 3–7 days. In sharp contrast, cough responses to these irritant gases were not affected by the treatment with the vehicle of MBP. These results suggest that the MBP-induced lingering elevation of cough responsiveness may be a contributing factor in the pathogenesis of chronic cough associated with eosinophilic infiltration of the airways.

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C/EBPβ and GATA-1 Synergistically Regulate Activity of the Eosinophil Granule Major Basic Protein Promoter: Implication for C/EBPβ Activity in Eosinophil Gene Expression

Blood ◽

10.1182/blood.v94.4.1429 ◽

1999 ◽

Vol 94 (4) ◽

pp. 1429-1439 ◽

Cited By ~ 47

Author(s):

Yuji Yamaguchi ◽

Hitoshi Nishio ◽

Kenji Kishi ◽

Steven J. Ackerman ◽

Toshio Suda

Keyword(s):

Binding Site ◽

Cell Line ◽

Binding Sites ◽

Basic Protein ◽

Regulatory Role ◽

Expression Vectors ◽

Regulatory Function ◽

Dependent Manner ◽

Major Basic Protein ◽

Eosinophil Granule

Abstract Eosinophil granule major basic protein (MBP) is expressed exclusively in eosinophils and basophils in hematopoietic cells. In our previous study, we demonstrated a major positive regulatory role for GATA-1 and a negative regulatory role for GATA-2 in MBP gene transcription. Further analysis of the MBP promoter region identified a C/EBP (CCAAT/enhancer-binding protein) consensus binding site 6 bp upstream of the functional GATA-binding site in the MBP gene. In the cell line HT93A, which is capable of differentiating towards both the eosinophil and neutrophil lineages in response to retinoic acid (RA), C/EBP mRNA expression decreased significantly concomitant with eosinophilic and neutrophilic differentiation, whereas C/EBPβ expression was markedly increased. Electrophoretic mobility shift assays (EMSAs) showed that recombinant C/EBPβ protein could bind to the potential C/EBP-binding site (bp −90 to −82) in the MBP promoter. Furthermore, we have demonstrated that both C/EBPβ and GATA-1 can bind simultaneously to the C/EBP- and GATA-binding sites in the MBP promoter. To determine the functionality of both the C/EBP- and GATA-binding sites, we analyzed whether C/EBPβ and GATA-1 can stimulate the MBP promoter in the C/EBPβ and GATA-1 negative Jurkat T-cell line. Cotransfection with C/EBPβ and GATA-1 expression vectors produced a 5-fold increase compared with cotransfection with the C/EBPβ or GATA-1 expression vectors individually. In addition, GST pull-down experiments demonstrated a physical interaction between human GATA-1 and C/EBPβ. Expression of FOG (F̲riendo̲fG̲ATA), which binds to GATA-1 and acts as a cofactor for GATA-binding proteins, decreased transactivation activity of GATA-1 for the MBP promoter in a dose-dependent manner. Our results provide the first evidence that both GATA-1 and C/EBPβ synergistically transactivate the promoter of an eosinophil-specific granule protein gene and that FOG may act as a negative cofactor for the eosinophil lineage, unlike its positively regulatory function for the erythroid and megakaryocyte lineages.

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Low-angle shadowing and colloidal gold-labeling of liposome interaction with the eosinophil major basic protein

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100123945 ◽

1992 ◽

Vol 50 (1) ◽

pp. 708-709

Author(s):

Margaret J. Hukee ◽

Randa I. Abu-Ghazaleh ◽

Franklyn G. Prendergast

Keyword(s):

Lipid Bilayers ◽

Mammalian Cells ◽

Plasma Membranes ◽

Basic Protein ◽

Direct Interaction ◽

Major Basic Protein ◽

Eosinophil Granule ◽

Spectroscopy Studies ◽

Eosinophil Major Basic Protein

The eosinophil major basic protein (MBP) has been localized in the crystalloid core of the specific eosinophil granule. High levels of MBP are present in sputa and at sites of epithelial damage in patients with brochial asthma suggesting that MBP is released from the eosinophil. In vitro, the toxicity of MBP to mammalian cells, helminths, protozoa, and bacteria, has been demonstrated. MBP also induced the degranulation of platelets, basophils, and mast cells. Since these events involve extracellular MBP and rupture of plasma membranes, the mechanism of toxicity may be due to a direct interaction between MBP and biological membranes. Fluorescence spectroscopy studies using synthetic lipid bilayers (liposomes) showed the ability of MBP to induce disorder, fusion and lysis of those membranes. In this report, structural evidence is presented for liposome aggregation and possible lysis in the presence of MBP.

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Activation of platelets by eosinophil granule proteins.

Journal of Experimental Medicine ◽

10.1084/jem.172.4.1271 ◽

1990 ◽

Vol 172 (4) ◽

pp. 1271-1274 ◽

Cited By ~ 114

Author(s):

M S Rohrbach ◽

C L Wheatley ◽

N R Slifman ◽

G J Gleich

Keyword(s):

Basic Protein ◽

Major Basic Protein ◽

Cationic Proteins ◽

Distinct Mechanism ◽

Eosinophil Peroxidase ◽

Eosinophil Granule Proteins ◽

Granule Proteins ◽

Eosinophil Granule ◽

Dose Dependent

Two of the four principal cationic proteins of the eosinophil granule, major basic protein (MBP) and eosinophil peroxidase (EPO), were shown to be platelet agonists. Both MBP and EPO evoked a dose-dependent nonlytic secretion of platelet 5-hydroxytryptamine in unstirred platelet suspensions even in the presence of 10 microM indomethacin. MBP also evoked secretion of platelet alpha granule and lysosome components. Secretion by MBP and EPO was inhibited by 1 microM PGE1, but the nature of the inhibition differed from that observed with thrombin. Thus, MBP and EPO can be classified as strong platelet agonists with a distinct mechanism of activation.

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C/EBPβ and GATA-1 Synergistically Regulate Activity of the Eosinophil Granule Major Basic Protein Promoter: Implication for C/EBPβ Activity in Eosinophil Gene Expression

Blood ◽

10.1182/blood.v94.4.1429.416k13_1429_1439 ◽

1999 ◽

Vol 94 (4) ◽

pp. 1429-1439 ◽

Cited By ~ 1

Author(s):

Yuji Yamaguchi ◽

Hitoshi Nishio ◽

Kenji Kishi ◽

Steven J. Ackerman ◽

Toshio Suda

Keyword(s):

Binding Site ◽

Cell Line ◽

Binding Sites ◽

Basic Protein ◽

Regulatory Role ◽

Expression Vectors ◽

Regulatory Function ◽

Dependent Manner ◽

Major Basic Protein ◽

Eosinophil Granule

Eosinophil granule major basic protein (MBP) is expressed exclusively in eosinophils and basophils in hematopoietic cells. In our previous study, we demonstrated a major positive regulatory role for GATA-1 and a negative regulatory role for GATA-2 in MBP gene transcription. Further analysis of the MBP promoter region identified a C/EBP (CCAAT/enhancer-binding protein) consensus binding site 6 bp upstream of the functional GATA-binding site in the MBP gene. In the cell line HT93A, which is capable of differentiating towards both the eosinophil and neutrophil lineages in response to retinoic acid (RA), C/EBP mRNA expression decreased significantly concomitant with eosinophilic and neutrophilic differentiation, whereas C/EBPβ expression was markedly increased. Electrophoretic mobility shift assays (EMSAs) showed that recombinant C/EBPβ protein could bind to the potential C/EBP-binding site (bp −90 to −82) in the MBP promoter. Furthermore, we have demonstrated that both C/EBPβ and GATA-1 can bind simultaneously to the C/EBP- and GATA-binding sites in the MBP promoter. To determine the functionality of both the C/EBP- and GATA-binding sites, we analyzed whether C/EBPβ and GATA-1 can stimulate the MBP promoter in the C/EBPβ and GATA-1 negative Jurkat T-cell line. Cotransfection with C/EBPβ and GATA-1 expression vectors produced a 5-fold increase compared with cotransfection with the C/EBPβ or GATA-1 expression vectors individually. In addition, GST pull-down experiments demonstrated a physical interaction between human GATA-1 and C/EBPβ. Expression of FOG (F̲riendo̲fG̲ATA), which binds to GATA-1 and acts as a cofactor for GATA-binding proteins, decreased transactivation activity of GATA-1 for the MBP promoter in a dose-dependent manner. Our results provide the first evidence that both GATA-1 and C/EBPβ synergistically transactivate the promoter of an eosinophil-specific granule protein gene and that FOG may act as a negative cofactor for the eosinophil lineage, unlike its positively regulatory function for the erythroid and megakaryocyte lineages.

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Role of leukemia inhibitor factor (LIF) in decidualisation of murine uterine stromal cells in vitro

Journal of Endocrinology ◽

10.1677/joe.0.1810477 ◽

2004 ◽

Vol 181 (3) ◽

pp. 477-492 ◽

Cited By ~ 19

Author(s):

AA Fouladi Nashta ◽

CV Andreu ◽

N Nijjar ◽

JK Heath ◽

SJ Kimber

Keyword(s):

In Vitro Culture ◽

Stromal Cells ◽

Control Level ◽

Calf Serum ◽

Prostaglandin E ◽

Dependent Manner ◽

Alp Activity ◽

Dose Dependent ◽

In Cells

Decidualisation of uterine stromal cells is a prerequisite for implantation of the embryo in mice. Here we have used an in vitro culture system in which stromal cells decidualise as indicated by a number of markers, including an increase in alkaline phosphatase (ALP) activity. The latter was used as a quantitative marker of decidualisation in the presence of low (2%) fetal calf serum. Prostaglandin E(2) (PGE(2)), which is known to induce decidualisation, increased ALP activity, and this effect was blocked in a dose-dependent manner by indomethacin. Leukemia inhibitory factor (LIF) was then examined, but it had no effect on PGE(2) secretion. However, LIF suppressed ALP activity in a dose-dependent manner in the presence of 2% serum, while an inhibitor of LIF that competes for binding to its receptor reversed the effect of LIF and increased ALP activity above the control level. In serum-free cultures, stromal cells differentiated rapidly, and no differences were observed between LIF-treated and untreated cultures. Stromal cells produce LIF during in vitro culture, and this peaked at 48 h. Freshly collected stromal cells from both day-2 and -4 pregnant mice expressed mRNA for the LIF receptor, and the transcript level was higher in cells isolated on day 4. However, no differences were observed in the relative levels of transcripts in cells from day 2 and day 4 after culture, nor were there differences between the LIF-treated cultures and controls. Therefore, in this study, we have shown that LIF suppresses decidualisation of murine uterine stromal cells in the presence of serum, this is not due to the regulation of PGE(2) secretion by stromal cells.

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Biochemical and amino acid sequence analysis of human eosinophil granule major basic protein.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)37791-3 ◽

1988 ◽

Vol 263 (25) ◽

pp. 12559-12563

Author(s):

T L Wasmoen ◽

M P Bell ◽

D A Loegering ◽

G J Gleich ◽

F G Prendergast ◽

...

Keyword(s):

Amino Acid ◽

Sequence Analysis ◽

Amino Acid Sequence ◽

Basic Protein ◽

Major Basic Protein ◽

Amino Acid Sequence Analysis ◽

Human Eosinophil ◽

Eosinophil Granule

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A Novel and Highly Divergent Homolog of Human Eosinophil Granule Major Basic Protein

Journal of Biological Chemistry ◽

10.1074/jbc.274.20.14464 ◽

1999 ◽

Vol 274 (20) ◽

pp. 14464-14473 ◽

Cited By ~ 54

Author(s):

Douglas A. Plager ◽

David A. Loegering ◽

Deborah A. Weiler ◽

James L. Checkel ◽

Jill M. Wagner ◽

...

Keyword(s):

Basic Protein ◽

Major Basic Protein ◽

Human Eosinophil ◽

Eosinophil Granule

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Normal Saline Solution Nasal-Pharyngeal Irrigation Improves Chronic Cough Associated with Allergic Rhinitis

American Journal of Rhinology and Allergy ◽

10.2500/ajra.2017.31.4418 ◽

2017 ◽

Vol 31 (2) ◽

pp. 96-104 ◽

Cited By ~ 6

Author(s):

Lin Lin ◽

Zhongchun Chen ◽

Yitan Cao ◽

Guangbin Sun

Keyword(s):

Allergic Rhinitis ◽

House Dust ◽

House Dust Mite ◽

Chronic Cough ◽

Saline Solution ◽

Basic Protein ◽

Leukotriene C4 ◽

Major Basic Protein ◽

Leicester Cough Questionnaire ◽

Dust Mite

Background Upper airway inflammation is one of the most commonly identified causes of chronic cough, although the underlying mechanism is not clear. This study compared normal saline solution nasal-pharyngeal irrigation (NSNPI) and fluticasone propionate nasal spray (FPNS) treatment for chronic cough associated with allergic rhinitis (AR). Methods Patients with suspected AR to house-dust mite were enrolled, and the symptom of cough was assessed by a cough symptom score and the Leicester Cough Questionnaire, and cough response to capsaicin was evaluated. AR was assessed by using the visual analog scale (VAS) and the Mini Juniper Rhinoconjunctivitis Quality of Life Questionnaire (MiniRQLQ). Mediators, including histamine, leukotriene C4, and prostaglandin D2, and the major basic protein from nasal lavage fluid (NLF) were examined. The patients were treated with NSNPI (the NSNPI group) or FPNS (the FPNS group) for 30 days, after which they were reassessed. Results Forty-five of 50 patients completed this study. The scores of the cough symptom and the Leicester Cough Questionnaire, and the capsaicin cough threshold all improved statistically after NSNPI but did not change after FPNS. There were statistically significant changes in the evaluations of the MiniRQLQ and the mediators, including histamine and leukotriene C4, in the NLF in the NSNPI group. However, significant changes were found in the assessments of VAS, MiniRQLQ, and all above mediators including histamine, leukotriene C4, and prostaglandin D2, and the major basic protein in the NLF of the FPNS group. Furthermore, the assessments of VAS and all the mediators were reduced more in the FPNS group compared with those in the NSNPI group. Conclusion The patients with suspected AR to house-dust mite reported a better relief of the cough symptom after 30 days of treatment with NSNPI compared with that after nasal corticosteroid.

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