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Respirology (Carlton, Vic.) Aug 2009Attacks of fatal asthma have been shown to be either of short duration or long duration and associated with differing degrees of smooth muscle contraction, luminal mucus... (Comparative Study)
Comparative Study
BACKGROUND AND OBJECTIVE
Attacks of fatal asthma have been shown to be either of short duration or long duration and associated with differing degrees of smooth muscle contraction, luminal mucus deposition and ratios of eosinophils to neutrophils in the airway wall. We hypothesized that this bimodal distribution might be related to airway mast cell degranulation.
METHODS
Airway sections from cases of fatal asthma in the second Victorian asthma mortality study and from cases coming to coronial autopsy in Perth were examined. Tryptase-positive mast cells, numbers of intact and degranulated mast cells, post-mortem blood salbutamol levels and airway dimensions were measured.
RESULTS
Although the total number of mast cells were similar, the ratio of degranulated to intact mast cells (D/I) was significantly increased in the smooth muscle (P < 0.05) and outer airway wall (P < 0.001), in short-duration cases compared with long-duration cases. Proportional muscle shortening was significantly increased (P < 0.05) in short-duration cases (17 +/- 7%) compared with long-duration cases (11 +/- 7%). Blood salbutamol levels were related to the total airway wall mast cell D/I ratio for all cases combined (r = 0.57, P = 0.01).
CONCLUSIONS
The duration of a fatal attack of asthma may be partly determined by the degranulation of mast cells.
Topics: Adolescent; Adult; Asthma; Australia; Cell Degranulation; Child; Death; Disease Progression; Female; Humans; Lung; Male; Mast Cells; Middle Aged; Muscle Contraction; Muscle, Smooth; Predictive Value of Tests; Prognosis; Risk Factors; Severity of Illness Index; Time Factors; Young Adult
PubMed: 19659839
DOI: 10.1111/j.1440-1843.2009.01551.x -
PloS One 2024Research on neutrophil biology has been limited by the short life span and limited genetic manipulability of these cells, driving the need for representative and...
Research on neutrophil biology has been limited by the short life span and limited genetic manipulability of these cells, driving the need for representative and efficient model cell lines. The promyelocytic cell line HL-60 and its subline PLB-985 can be differentiated into neutrophil-like cells (NLCs) and have been used to study neutrophil functions including chemotaxis, phagocytosis, endocytosis, and degranulation. Compared to neutrophils derived from hematopoietic stem cells, NLCs serve as a cost-effective neutrophil model. NLCs derived from both HL-60 and PLB-985 cells have been shown to perform degranulation, an important neutrophil function. However, no study has directly compared the two lines as models for degranulation including their release of different types of mobilizable organelles. Furthermore, Nutridoma, a commercially available supplement, has recently been shown to improve the chemotaxis, phagocytosis, and oxidative burst abilities of NLCs derived from promyelocytic cells, however it is unknown whether this reagent also improves the degranulation ability of NLCs. Here, we show that NLCs derived from both HL-60 and PLB-985 cells are capable of degranulating, with each showing markers for the release of multiple types of secretory organelles, including primary granules. We also show that differentiating HL-60 cells using Nutridoma does not enhance their degranulation activity over NLCs differentiated using Dimethyl Sulfoxide (DMSO) plus Granulocyte-colony stimulating factor (G-CSF). Finally, we show that promyelocytic cells can be genetically engineered and differentiated using these methods, to yield NLCs with a defect in degranulation. Our results indicate that both cell lines serve as effective models for investigating the mechanisms of neutrophil degranulation, which can advance our understanding of the roles of neutrophils in inflammation and immunity.
Topics: Humans; Neutrophils; HL-60 Cells; Phagocytosis; Cell Differentiation; Granulocyte Precursor Cells; Cell Degranulation
PubMed: 38324578
DOI: 10.1371/journal.pone.0297758 -
Italian Journal of Anatomy and... 2001Brain mast cells are selectively concentrated in the thalamus of many mammalian species. We here describe by light and electron microscopy in the normal thalamus of...
Brain mast cells are selectively concentrated in the thalamus of many mammalian species. We here describe by light and electron microscopy in the normal thalamus of adult rats the features of mast cell degranulation, which indicate an active release of the mediators stored in their intracellular granules. The state of activity of thalamic mast cells in basal conditions was found to range from the release of a few granules to a massive degranulation, and the latter process was much less frequent than a partial degranulation. Mast cells were subdivided in three categories (fully granulated, partially or massively degranulated) on the basis of their cytoplasmic features revealed by acidic toluidine blue staining; the fully granulated cells were found to represent only 23 % of thalamic mast cells. This strategy of evaluation could be of help in the comparison of the functional correlates of mast cells in different conditions and experimental paradigms. However, we also demonstrated with image analysis a continuum of the variation of staining intensity of granulated and degranulating mast cells, without a sharp subdivision into different categories. Therefore our results reveal that the vast majority of mast cells are active in the thalamus in basal conditions, and that image analysis can provide an objective index of the activity of these cells.
Topics: Animals; Blood Vessels; Male; Mast Cells; Microscopy, Electron; Rats; Rats, Wistar; Secretory Vesicles; Thalamus
PubMed: 11729991
DOI: No ID Found -
European Journal of Cell Biology Sep 2009Measurement of released granule components, popularly used to quantify mast cell exocytosis, does not deliver real-time information about degranulation at the...
Measurement of released granule components, popularly used to quantify mast cell exocytosis, does not deliver real-time information about degranulation at the single-cell level nor the ratio of responding/non-responding cells. Rather it provides, only end-point, bulk-population data. Here we studied degranulation of rat peritoneal mast cells dispersed in a narrow horizontal channel between a silicon substrate and a glass plate. Upon exposure to a concentration gradient of a soluble stimulus, degranulation started from those cells facing towards the highest concentration of stimulus. We captured images of exocytosing cells without the need for phase-contrast or differential interference-contrast microscopy. This was achieved using the reflection caused by the silicon substrate. The time-lapse images of cells in the channel were segmented into multiple concentration belts to identify the proportion of degranulated cells in each belt region. Maximum ratios of degranulated cells in the belt regions determined by time-course curve fitting calculations were then plotted against the distance from the stimulus injection site, resulting in a sigmoidal response curve. This method provides a powerful means for real-time analysis of concentration- and stimulus-dependent degranulation of mast cells and allows comparison of cell responses under different conditions. To show its effectiveness, we evaluated the effect of a protein kinase C (PKC) inhibitor, Gö6976, on degranulation induced by various stimuli. In contrast to stimulation with concanavalin A+lysophosphatidylserine (lysoPS) or nerve growth factor+lysoPS (completely inhibited by Gö6976 over the whole range of stimulus concentrations used) or compound 48/80 and mastoparan (no inhibition by Gö6976), stimulation with ionomycin, a known Ca(2+) ionophore, showed a concentration-dependent inhibition by Gö6976, with a major inhibition at low stimulus concentrations and a diminished one at higher ionomycin concentrations. The results indicate that ionomycin-induced degranulation is mainly induced via a PKC-independent signal cascade at high stimulus concentrations, whereas below a certain concentration, degranulation is completely dependent on PKC.
Topics: Animals; Cell Degranulation; Glass; Image Processing, Computer-Assisted; Ionomycin; Mast Cells; Protein Kinase C; Rats; Silicon
PubMed: 19515452
DOI: 10.1016/j.ejcb.2009.03.004 -
Immunology May 1974With a modified rat mast cell degranulation (RMCD) technique developed by Korotzer, Haddad and Lopapa (1971), the mechanism of mast cell degranulation by IgE—anti-IgE...
With a modified rat mast cell degranulation (RMCD) technique developed by Korotzer, Haddad and Lopapa (1971), the mechanism of mast cell degranulation by IgE—anti-IgE reaction and the inhibitory effect of cAMP-related compounds upon IgE-mediated mast cell degranulation were studied. Degranulations of 90 per cent or more were decreased to 13–16 per cent when the mast cells were pretreated with human IgE or normal human serum. However, if rat mast cells were pretreated with anti-human IgE rabbit serum or normal rabbit serum, the degranulation per cent in these cells by IgE—anti-IgE reaction was the same as in the nontreated cells. These results suggest the presence of receptors in rat mast cells for human IgE or normal human serum, and the lack of receptors in these cells for anti-human IgE rabbit serum or normal rabbit serum. Treatment of isolated rat mast cells with adenyl cyclase stimulating agents (isoprenaline, adrenaline, prostaglandin E and E) and theophylline or aminophylline, which inhibit the enzymatic degradation of cAMP, also inhibited the morphological degranulation of the mast cells. Cromoglycate or chlorophenes in derivatives, which might have a stabilizing effect of the cell membrane, also inhibited the degranulation of the rat mast cells mediated by IgE—anti-IgE reaction. These results support the attractive hypothesis that cAMP occupies a central modulatory role in the mast cell degranulation by IgE—anti-IgE reaction.
Topics: Aminophylline; Animals; Antibodies, Anti-Idiotypic; Antigen-Antibody Complex; Binding Sites, Antibody; Blood; Cell Membrane; Cromolyn Sodium; Cyclic AMP; Epinephrine; Hypersensitivity, Immediate; Immune Sera; Immunoglobulin E; Isoproterenol; Male; Mast Cells; Prostaglandins; Rats; Theophylline
PubMed: 4368738
DOI: No ID Found -
Inflammation Mar 1981A method is described for the quantitative monitoring of human eosinophil degranulation using interference contrast microscopy. Using staphyloccoci as a stimulus,...
A method is described for the quantitative monitoring of human eosinophil degranulation using interference contrast microscopy. Using staphyloccoci as a stimulus, degranulated cells appeared larger than nondegranulating cells, were ameboid in shape and exhibited large nude areas of cytoplasm with prominent nuclei. Granules were observed to marginate along the plasma membrane and discharge into the exterior of the cell. Eosinophils that were not induced to degranulate were spherical in shape and the cytoplasm contained numerous granules that often obscured the nuclei. Pharmacological agents that increase intracellular cAMP prevented degranulation, whereas those that increase cGMP had no effect on degranulation. Colchicine inhibited degranulation but did not interfere with the phagocytosis of staphyloccoci. Endotoxin-activated serum, ECF-A, phytohemagglutinin, concanavalin A, levamisole, and compound 48/80 caused degranulation of eosinophils per se. The presence of disodium cromoglycate prevented this degranulation. Compound 48/80 and disodium cromoglycate had no effect on the level of intracellular cAMP and cGMP.
Topics: Chemotactic Factors; Colchicine; Cromolyn Sodium; Cyclic AMP; Cyclic GMP; Cytoplasmic Granules; Eosinophils; Humans; Microscopy, Interference; Mitogens; Phagocytosis; p-Methoxy-N-methylphenethylamine
PubMed: 6262236
DOI: 10.1007/BF00910778 -
The American Journal of Pathology Aug 1981The degranulation response of peritoneal mast cells to calcium ionophore A23187 and compound 48/80 has been compared by scanning electron microscopy and transmission... (Comparative Study)
Comparative Study
The degranulation response of peritoneal mast cells to calcium ionophore A23187 and compound 48/80 has been compared by scanning electron microscopy and transmission electron microscopy in normal C57 black mice and C57 beige mice with a genetic defect analogous to the human Chédiak-Higashi Syndrome (CHS). These methods reveal granule secretion in response to the degranulating agents in mast cells of both normal and beige mice. The observations indicate that beige peritoneal mast cells retain the capacity to exocytose their granules upon stimulation and suggest that the defect underlying the formation of the mega-inclusions is not attributable to impaired degranulation.
Topics: Animals; Ascitic Fluid; Calcimycin; Calcium; Chediak-Higashi Syndrome; Disease Models, Animal; Exocytosis; Female; Male; Mast Cells; Mice; Mice, Mutant Strains; Microscopy, Electron; Microscopy, Electron, Scanning; p-Methoxy-N-methylphenethylamine
PubMed: 6789683
DOI: No ID Found -
Reviews in the Neurosciences Oct 2017Mast cells are immunological cells that are diversely distributed in different parts of the body. Their role in various pathological conditions such as hypersensitivity,... (Review)
Review
Mast cells are immunological cells that are diversely distributed in different parts of the body. Their role in various pathological conditions such as hypersensitivity, atherosclerosis, pulmonary hypertension, and male infertility has been reported by different scientists. Apart from these, a number of studies have shown their important role in pathogenesis of neuropathic pain of diverse aetiology. They have been found to release active mediators, primarily histamine and serotonin on degranulation in response to different stimuli including chemical, nerve damage, toxin or disease-related conditions. The mast cells stabilizer has shown pain attenuating effects by preventing degranulation of mast cells. Similarly, compound 48/80 (first dose 200 μg/100 g and after 6-h interval, second dose of 500 μg/100 g) caused the degranulation of the accumulated endoneurial histamine and 5-HT antagonists have shown pain relieving effects by attenuating the effects of histamine and serotonin, respectively. On the other hand, the mast cell degranulator compound 48/80 has shown dual action depending on its time of administration. The present review discusses the critical role of mast cells in the generation and maintenance of neuropathic pain in experimental models.
Topics: Animals; Cell Degranulation; Histamine; Humans; Mast Cells; Neuralgia; p-Methoxy-N-methylphenethylamine
PubMed: 28688228
DOI: 10.1515/revneuro-2017-0007 -
Anatomy & Cell Biology Dec 2017Allergic diseases are a significant health concern in developing countries. Type-A procyanidin polyphenols from cinnamon ( Blume) bark (TAPP-CZ) possesses antiasthmatic...
Allergic diseases are a significant health concern in developing countries. Type-A procyanidin polyphenols from cinnamon ( Blume) bark (TAPP-CZ) possesses antiasthmatic and antiallergic potential. The present study was aimed at the possible anti-allergic mechanism of TAPP-CZ against the compound 48/80 (C48/80)-induced mast cell degranulation in isolated rat peritoneal mast cells (RPMCs). TAPP-CZ (1, 3, 10, and 30 µg/ml) was incubated for 3 hours with isolated, purified RPMCs. The C48/80 (1 µg/ml) was used to induce mast cell degranulation. The mast cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay whereas histamine, β-hexosaminidase (β-HEX), and interleukin-4 (IL-4) levels were determined in RPMCs. TAPP-CZ (3, 10, and 30 µg/ml) showed significant and dose-dependent decrease in a number of degranulated cells and levels of markers (histamine, β-HEX, and IL-4) as compared with C48/80 control. In conclusion, TAPP-CZ stabilizes mast cell and cause inhibition of the allergic markers such as histamine, IL-4, and β-HEX in IgE-mediated manner. The present study supports mast cell stabilization as a possible mechanism of action of TAPP-CZ against immune respiratory disorders such as asthma and allergic rhinitis.
PubMed: 29354299
DOI: 10.5115/acb.2017.50.4.275 -
Cell and Tissue Research Dec 2005The purpose of this study was to investigate the effect of oxygen-glucose deprivation (OGD) on degranulation and histamine release of mast cells. Cultured mast cells...
The purpose of this study was to investigate the effect of oxygen-glucose deprivation (OGD) on degranulation and histamine release of mast cells. Cultured mast cells were exposed to OGD for 1, 2, 4, 8, or 16 h. At 2 h of OGD exposure, the degranulation percentage of mast cells had increased and subsequently showed a progressive further increase, associated with a similar change in lactate dehydrogenase release. Histamine release increased significantly from 1 h of OGD exposure. These results indicate that OGD induces mast cells to degranulate, possibly via a cytotoxic response. This in vitro ischemic model of mast cells might clarify their roles in the pathological processes induced by cerebral ischemia.
Topics: Animals; Cell Degranulation; Cell Hypoxia; Glucose; Histamine Release; L-Lactate Dehydrogenase; Male; Mast Cells; Oxygen; Peritoneal Cavity; Rats; Rats, Sprague-Dawley
PubMed: 16133147
DOI: 10.1007/s00441-005-0041-z