Histamine release from rat mast cells sensitized with mouse antiserum

The characteristics of the antigen-induced and non-antigen-induced histamine release from rat peritoneal mast cells sensitized in vitro with mouse anti-ovalbumin serum were investigated. The effects of some antiallergic drugs on these release reactions were also studied. Besides antigen-specific IgE antibody, heat-labile factor(s) responsible for the non-antigen-induced histamine release were found in mouse antiserum. Such factors were also present in normal mouse serum. In the absence of antigen, the combination of phosphatidyl serine and Ca++ induced some extent of histamine release from mast cells treated with these factors. From the present results it is suggested that quercetin selectively and verapamil primarily act to block calcium-gate opening resulting from antigen-antibody interaction on the mast cell membrane, while theophylline and disodium cromoglycate selectively inhibit the passage of calcium through open calcium channels.     

Heterogeneity of calcium channels in mast cells and basophils and the possible relevance to pathophysiology of lung diseases: a review

Calcium plays a critical role in the formation and secretion of a wide variety of chemical mediators. Calcium slow-channel blockers, e.g. nifedipine and verapamil, have been shown to inhibit the synthesis of SRS (SRS-A, leukotrienes) in human and guinea pig lung tissue, thromboxane A2 formation in rat lung and platelet activating factor in human neutrophils. Verapamil and nifedipine also prevent the release of lysosomal enzymes from rabbit and human polymorphonuclear neutrophils. Calcium-channel blockers produce variable inhibitory effects on allergic and nonallergic histamine secretion. Ca++-entry blockers also inhibit the Ca++ uptake (influx) into mast cells. Many of these inhibitory effects of Ca++ antagonists are antagonized by an increased extracellular Ca++ ion concentration. The magnitude of the inhibitory influences of Ca++-channel blockers on allergic and nonallergic release of chemical mediators appears to depend on the cell source, species, nature and the concentration of the secretory stimuli as well as on the composition and pH of buffers and the concentration of Ca++-entry blockers used. The data summarized in this review suggest the existence of a functional heterogeneity of Ca++ channels in leukocytes, mast cells and basophils. Interference with the Ca++-dependent steps involved in the formation and/or release of chemical mediators appears to be the primary mode of action for Ca++-channel blockers in these cells. The differential effects of Ca++ antagonists on Ca++-dependent activation of phospholipase A2,5-lipoxygenase, and calmodulin (or other intracellular Ca++-binding proteins) in different cell types (mast cells, basophils, leukocytes, lung tissue, etc.) may explain the variation of their effectiveness in inhibiting the synthesis/release of chemical mediators and antagonizing bronchoconstriction in response to diverse stimuli. During the process of hypersensitization and in immediate hypersensitivity diseases, Ca++ homeostasis (uptake, mobilization, distribution, relocation, etc.) may be altered in leukocytes (mast cells, basophils) and lung tissues. The altered Ca++ homeostasis could be responsible for the induction of airway hyperreactivity in asthmatics and for hyperreleasability of chemical mediators from leukocytes, mast cells and other cell types.(ABSTRACT TRUNCATED AT 250 WORDS)     

Role of calcineurin in the regulation of human lung mast cell and basophil function by cyclosporine and FK506

BACKGROUND AND PURPOSE: Cyclosporine and FK506 are thought to act by targeting the Ca2+-dependent protein phosphatase, calcineurin. The aim of the present study was to determine whether cyclosporine and FK506 stabilize mast cells and basophils by interacting with calcineurin. EXPERIMENTAL APPROACH: The effects of cyclosporine and FK506 on the IgE-mediated release of histamine from mast cells and basophils were evaluated. The presence of calcineurin in cells was determined by Western blotting. Ca2+-dependent protein phosphatase activities were assessed in cell extracts using a synthetic phosphorylated peptide that is known to serve as a substrate for calcineurin. KEY RESULTS: FK506 was about 100-fold more potent than cyclosporine as an inhibitor of IgE-dependent histamine release from mast cells and basophils. Immunoblotting of solubilized preparations of purified cells demonstrated the presence of calcineurin in mast cells and basophils. In enzyme assays, mast cells expressed approximately 7-fold higher Ca2+-dependent protein phosphatase activity than basophils. Whereas cyclosporine effectively inhibited Ca2+-dependent protein phosphatase activity in cell extracts, FK506 was considerably less effective. CONCLUSIONS AND IMPLICATIONS: FK506 and cyclosporine inhibit the stimulated release of histamine from mast cells and basophils. However, the ability of cyclosporine, but not FK506, to inhibit Ca2+-dependent protein phosphatase activity questions whether FK506 stabilizes mast cells and basophils by interacting with calcineurin.     

Substance P-induced histamine release from rat peritoneal mast cells and its inhibition by antiallergic agents and calmodulin inhibitors.

Substance P-induced histamine release and Ca2+ release from the intracellular Ca store of rat peritoneal mast cells were inhibited by both antiallergic drugs and microtubule inhibiting agents. It was found that in the case of antiallergic compounds, histamine release inhibition may be intimately related to the inhibition of Ca2+ release from the intracellular store in which the microtubules play an important role. When mast cells were pretreated with either theophylline or dibutyryl cAMP, the inhibition of histamine release was closely related to the inhibition of Ca2+ release from the intracellular Ca store. Calmodulin inhibitors were also effective in inhibiting histamine release from mast cells induced by substance P. The inhibitory potencies of calmodulin inhibitors on histamine release from mast cells were closely correlated with those exerted on calmodulin activity.     

Cyclic AMP independent inhibition by papaverine of histamine release induced by compound 48/80.

Compound 48/80-induced histamine release from isolated rat peritoneal mast cells was inhibited in a dose-dependent manner by papaverine (ic₅₀ approx 20 μM). This effect of papaverine was not influenced by PGE₁ (14–140 μM), even though PGE₁ markedly increased must cell cAMP levels. Papaverine (0.5 mM) completely inhibited histamine release without causing any change in cAMP levels. Theophylline (0.1 and 0.5 mM) potentiated histamine release induced by submaximal concentrations of compound 48/80, while cAMP levels were increased. IBM X was as potent as papaverine in causing inhibition of mast cell phosphodiesterase. IBM X (0.14–0.7 mM) had no effect on histamine release but caused a 6–20 fold increase in mast cell cyclic AMP. Papaverine inhibition of histamine release was gradual at the onset and was parallelled by a depletion of mast cell ATP content. The inhibition of 48/80-induced histamine release and depletion of mast cell ATP levels was reversed by glucose. It is concluded that papaverine induced inhibition of 48/80-induced histamine release is independent of cAMP, is unrelated to phosphodiesterase inhibition but is dependent upon inhibition of energy production.     

Adenosine triphosphatase in non-secreting and secreting mast cells

A Ca++-Mg++ ATPase has been demonstrated in the plasma membrane of rat peritoneal mast cells. The enzyme is localized by electron microscopy on the outer surface of the membrane. This agrees with the biochemical findings. A Ca++-Mg++ activated ATPase has also been shown to be present in the granule membrane. The optimal pH of the plasma membrane enzyme is close to the optimal pH for the histamine release. All the 14 inhibitors of plasma membrane ATPase tested - which caused varying degrees of inhibition of the enzyme - also inhibited histamine release induced by antigen, compound 48/80 and the divalent ionophore A23187. The conclusion from the study with the inhibitors is that a mild inhibition of the enzyme is compatible with histamine release, but a pronounced inhibition of the enzyme is always associated with inhibition of histamine release. ATP in low concentrations potentiates the release.     

Inhibitory role of cyclic adenosine 3' 5'-monophosphate in histamine release from rat peritoneal mast cells in vitro.

Cyclic adenosine 3′,5′-monophosphate (cyclic AMP) accumulated in rat peritoneal mast cells during incubation with epinephrine and theophylline, correlating well with the inhibition of histamine release from the cells by these agents, but occurring on different time courses. During stimulation with epinephrine, the initial, rapid accumulation of radioactive cyclic AMP in mast cells previously labeled with radioactive adenine preceded the elevation of total cyclic AMP content and the increase in the inhibition of histamine release. Theophylline, on the other hand, rapidly elevated the cellular levels of cyclic AMP before the accumulation of radioactive cyclic AMP and the inhibition of histamine release became detectable. Cyclic AMP, dibutyryl cyclic AMP, epinephrine and theophylline were all effective in suppressing histamine release from mast cells and from isolated granules in isotonic sucrose solution, but were less effective or not effective in NaCl-containing media. Cyclic AMP also suppressed the extrusion of granules from mast cells induced by compound $\text{48}\text{80}$ in isotonic sucrose solution. These results indicate that the formation of cyclic AMP from special pools of ATP is required for inhibiting histamine release from mast cells, and also that cyclic AMP regulates histamine release by interfering in an early stage of the release process prior to the interaction of the granular amine with the extracellular cations.     

Histamine release from saponin-permeabilized rat mast cells by calcium

The first effect of receptor activation on the mast cell surface, initiating histamine secretion, is an increase in the cytosol Ca2+ concentration. It should then be possible to induce histamine secretion by calcium alone, if the calcium permeability of the cell membrane could be increased without any significant interference with the physiological cell functions. This was achieved in the present study by adding low concentrations of saponin (0.0005% and 0.001% w/v) to the medium. When calcium was added to the saponin-permeabilized cells, around 40% histamine release occurred with 0.25 mM extracellular calcium (free Ca2+ 0.15 mM). The release was inhibited by antimycin A (1 microM). Transmission electron microscopy showed formation of vacuoles containing granules stripped of their membranes, which characterize a secretory response. The observations are consistent with a limited increase in the calcium permeability of the cell membrane for a brief period. There was apparently an increase in the cytoplasmic calcium concentration, which acted through calmodulin, since the histamine release induced by calcium from the permeabilized mast cells could be inhibited by a calmodulin-antagonist, mepacrine (10-30 microM).     

Histamine release from rat neutrophils and mast cells as induced by ionophore--a pharmacological approach--.

A comparative study was carried out on the histamine release from rat neutrophils and mast cells by calcium ionohpore A 23 187 (Ionophore). A maximum release of histamine from neutrophils was induced by 10(-6) g/ml Ionophore and that from mast cells was 5 x 10(-6) g/ml. A fairly good correlation was found between the 45Ca incorporation into and the histamine release from both cells. The Ionophore-induced histamine release from both cells was decreased in Ca2+-free Tyrode's solution and by pretreatment with 0.05 M EDTA. Effects of different drugs on Ionophore-induced histamine release from neutrophils were similar to those seen in mast cells. Dibutyl cyclic adenosine monophosphate, theophylline, isoproterenol and prostaglandin E1 had not or only a slight inhibition on the release. The dose dependent inhibition of release was observed with disodium cromoglycate, N-(3',4'-dimethoxycinnamoyl) anthranilic acid and disodium baicalein phosphate, in experiments using both cells. Colchicine did not inhibit the reaction in these cells, however phosphatidylserine enhanced the reaction. On the other hand, the effect of concanavalin A was different in each type of cells, the release from mast cells was inhibited while the release from neutrophils was potentiated. These findings suggest the similarity of biochemical events in Ionophore-induced histamine release from neutrophils and mast cells.     

Change in the activity of the cyclic AMP-dependent protein kinase in antigen-stimulated sensitized mast cells and effect of drugs inhibiting allergic mediator release

The activity of cyclic AMP-dependent protein kinase (protein kinase A) in the sensitized rat mast cell was decreased 2 min after antigen challenge when the histamine release exhibited a maximum. Drugs inhibiting allergic mediator release such as disodium cromoglycate, tranilast and theophylline significantly inhibited antigen-induced histamine release and reduced a decrease in the activity of protein kinase A. These results suggest that protein kinase A is involved in the histamine releasing process in the mast cell, and drugs inhibiting allergic mediator release cause their effects partially through the inhibition of protein kinase A.     

The role of membrane bound sialic acid of rat mast cells in histamine release induced by compound 48/80 and derivatives as well as calcium

Histamine release induced by compound 48/80 from rat mast cells is not dependent on extracellular Ca2+. Preincubation of mast cells with trypsin has only little effects on histamine release induced by this polycation. This work also demonstrates that histamine release induced by compound 48/80 and its analogues in the absence of extracellular Ca2+ depends on membrane bound sialic acid of the mast cell. Neuraminidase treatment of the cells in the presence of extracellular Ca2+ leads to histamine liberation. These findings suggest that sialic acid residues of the mast cell membrane constitute the site at which polycations exert their stimulatory actions of histamine liberation.     

Interaction between barium, strontium and calcium in histamine release by compound 48/80.

Pre-incubating rat peritoneal and pleural mast cells for 30 min at 2 degrees C in Ca2+-free Tyrode containing Ba2+ (30 mM) markedly reduced histamine release by compound 48/80 (0.5 microgram/ml). Evoked secretion was not significantly altered by pre-incubation with Sr2+ (30 mM) at this temperature whilst Mg2+ (30 mM) and La3+ (0.1 mM) slightly inhibited and potentiated release respectively. The effect of Ba2+ was concentration related and progressively reversed by Ca2+ (1--10 mM). Pre-incubation at 37 degrees C in Ca2+-free medium alone substantially reduced histamine release by compound 48/80. This reduction was unaltered by either Mg2+ (30 mM) or La3+ (0.1 mM), prevented by either Sr2+ (30 mM) or Ca2+ (1 mM) and augmented by Ba2+ (30 mM) which also markedly increased spontaneous histamine release during pre-incubation at 37 degrees C. Results suggest that Ba2+ and Sr2+ interact with cell-fixed calcium to modulate histamine release by compound 48/80.     

Roles of intracellular Ca<Superscript>2+</Superscript> and cyclic AMP in mast cell histamine release induced by radiographic contrast media

Mast cell histamine release is considered to be associated with the etiology of anaphylactoid reactions to iodinated radiographic contrast media (RCM). In the present study, the effects of various ionic and non-ionic RCM on histamine release from mast cells were compared, and the possible mechanisms of the histamine release were subsequently determined. Both ionic (ioxaglate and amidotrizoate) and non-ionic (iohexol, ioversol, iomeprol, iopamidol and iotrolan) RCM increased histamine release from the dissociated rat pulmonary cells, whereby ionic materials were more potent than non-ionic agents. There was no significant correlation between the extent of histamine release and the osmolarity of each RCM solution. In addition, hyperosmotic mannitol solution (1000 mOsm/kg) caused no marked histamine release. Thus, it is unlikely that the hyperosmolarity of RCM solutions contributes to the histamine release. RCM also stimulated, but to a lesser extent, the histamine release from rat peritoneal cells. The RCM-induced histamine release from both types of cells was inhibited by dibutyl cyclic AMP or combined treatment with forskolin and 3-isobutyl-1-methylxanthine. Corresponding to these results, RCM markedly reduced the cellular cyclic AMP content. On the other hand, the removal of intracellular but not the extracellular Ca2+ attenuated the RCM-induced mast cell histamine release. From these findings, it is suggested that the decrease in cellular cyclic AMP content and an increase in intracellular Ca2+ contribute at least in part to the RCM-induced mast cell histamine release.     

Essential role of ATP and possibility of activation of protein kinase C in Ca(2+)-dependent histamine release from permeabilized rat peritoneal mast cells.

To elucidate the role of ATP in histamine release, the present study was performed using beta-escin-permeabilized rat peritoneal mast cells. Ca(2+)-induced histamine release from permeabilized cells is totally dependent upon exogenous ATP in the medium. In the presence of Ca2+, ATP caused histamine release concentration-dependently at concentrations ranging from 0.01 to 5 mmol/l. The maximum release was achieved at 3 mmol/l of ATP in the medium. When the other adenosine nucleotides (AMP, ADP), or nonhydrolyzable ATP analogues (adenylylimidodiphosphate, beta, gamma-methylene ATP) were added in place to ATP, no histamine release took place. Other ribonucleoside triphosphates (GTP, ITP, UTP and CTP) had little effect at the same concentration range. When the ribonucleoside triphosphate content of mast cells was determined by means of HPLC, ITP and CTP were not detectable. A millimolar range of the ATP content was determined in mast cells, but the amounts of other ribonucleoside triphosphates (GTP and UTP) were remarkably lower than that of ATP. These results seem to indicate that the ATP molecule plays a crucial role in histamine release from rat mast cells in association with its concurrent hydrolysis. Furthermore, 12-O-tetradecanoylphorbol-13-acetate and 1-oleoyl-2-acetylglycerol enhanced histamine release elicited in the presence of Ca2+ (0.1 mumol/l) and ATP (3 mmol/l). Calphostin C, a potent inhibitor of protein kinase C, inhibited Ca2+/ATP-dependent histamine release by approximately 60%. At the same concentration, calphostin C inhibited by 95% protein kinase C activity in the crude extract obtained from rat mast cells. It was suggested that protein kinase C activation took place in the Ca2+/ATP-dependent histamine release from permeabilized rat mast cells.     

Inhibition of histamine release and ionophore-induced calcium flux in rat mast cells by lidocaine and chlorpromazine.

We studied the effects of lidocaine (L) and chlorromazine (C), two compounds known to affect the binding of calcium to cell membranes, on histamine release and calcium uptake by purified mast cells upon challenge with the ionophore A23,187 or with compound 48/80. At low concentrations L and C inhibited the Ca++ flux as well as histamine release while higher concentration caused enhancement in this function. Evidence was obtained that L 10(-4) M may displace Ca++ from the cell membranes.     

Synthetic polycations, polyethylenimines and polyallylamines release histamine from rat mast cells

The effects of synthetic polycations, which induce liposomal membrane fusion without inducing permeability changes, on histamine release from rat mast cells were investigated. Polyethylenimines and polyallylamines with various molecular weights released histamine from mast cells. Acetylated derivatives and triethylentetramine did not release histamine or serotonin from the cells. The histamine release induced by 10 micrograms/ml polyethylenimine with a molecular weight of 600 was inhibited by 1 mM dibutyryl cyclic AMP, but not by 1 MM 8-bromo cyclic GMP; 100 microM D-600, a calcium antagonist; or 30 microM W-7, a calmodulin inhibitor. In the presence of polyethylenimines with molecular weights of 600, 1,200 and 1,800, no detectable release of cytosolic lactate dehydrogenase was observed, indicating that histamine release induced by these polycations was not due to their cytotoxicity. The potencies of these polymers in inducing histamine release depended on their charges, but not on their degrees of polymerization. On the other hand, the actions of polyethylenimine with a molecular weight of 10,000 and polyallylamines with molecular weights of 3,000-4,000 and 10,000 in releasing lactate dehydrogenase were somewhat cytotoxic. These polycations did not induce serotonin release from rat platelets, suggesting that platelets have no coupling system of signal transduction by these polycations. Thus polycations seemed to interact with the mast cell membrane to induce histamine release, and the potencies of these polycations on mast cells seemed to differ from those of their effects on liposomes, which were examined previously.     

Ca(2+)-induced translocation of protein kinase C during Ca(2+)-dependent histamine release from beta-escin-permeabilized rat mast cells.

When rat mast cells were cultured for a short period in plastic dishes and adhering cells were permeabilized with beta-escin and exposed to Ca2+ at concentrations higher than 10(-7) mol/l, histamine release was induced dose-dependently. Protein kinase C (PKC) activity in the crude extracts obtained from adhering mast cells was induced in the presence of Ca2+, phospholipid and diacylglycerol. The apparent Km value of PKC for Ca2+ was 0.33 mumol/l, and this Ca2+ concentration was equivalent to that which can elicit half the maximum of the Ca(2+0-induced histamine release. After permeabilization, approximately 80% of the total PKC activity remained in the cytosolic fraction. In the resting state, 95% of the total PKC activity was detected in the soluble fraction, and the rest was detected in the membrane fraction. When permeabilized mast cells were incubated with Ca2+ at micromolar concentrations, which are effective in releasing histamine, the total PKC activity did not change. However, the translocation of PKC took place from the cytosolic fraction to the membrane fraction, corresponding to Ca2+ concentrations in the medium. When the crude PKC extract of mast cells was incubated with phospholipid vesicles and centrifuged, the PKC activity in the supernatant was diminished; the amount of PKC binding to the vesicles was dependent upon Ca2+ concentrations in the medium. Calphostin C, a potent PKC inhibitor, interacts with PKC in a noncompetitive manner, and it does not inhibit Ca(2+)-induced translocation of PKC. It can be concluded that PKC is translocated into the cell membrane along with an increase in intracellular Ca2+ concentrations and the subsequent activation of PKC may be crucial for the process leading to histamine release.     

The action of local anaesthetics on histamine release.

The effect of the local anaesthetics lidocaine, procaine and tetracaine on compound 48/80-induced histamine release from isolated rat mast cells has been investigated. They inhibited histamine release in a dose-dependent manner; at a concentration of 20 mM there was almost total inhibition of histamine release by lidocaine and about 75% inhibition by procaine. Tetracaine exerted a biphasic effect: at concentrations below 1 mM it inhibited, but at concentrations above 1 mM it potentiated histamine release. The inhibitory effect of lidocaine on compound 48/80-evoked histamine release was dependent upon the time of preincubation of mast cells with this anaesthetic and it persisted after washing the cells and resuspension in a lidocaine-free medium. An increase of calcium ions antagonized the inhibitory action of lidocaine. These results can be explained by (1) blockade of membrane receptors for calcium binding which leads to a decrease in intracellular calcium concentration and (2) increase of cellular cyclic AMP content which subsequently inhibits the releasing process.     

Induction of histamine release from rat peritoneal mast cells by histatins.

Human salivary histatins (Hsts), which belong to a salivary polypeptide family, have potent antifungal activity against Candida albicans and Cryptococcus neoformans, and are expected to be useful as therapeutic reagents against Candida species. However, little is known about the effect of Hsts on host immune systems. Thus we conducted a series of in vitro experiments with rat mast cells to determine whether histatin 5 (Hst 5) or histatin 8 (Hst 8) has a histamine-releasing effect on mast cells. Both Hst 5 and Hst 8 induced histamine release from rat peritoneal mast cells in a dose-dependent manner (10(-9) to 10(-5) M). Hst 5 had a stronger releasing effect than Hst 8. The histamine release induced by Hst 5 (10(-6) M) was increased by the presence of 0.5 mM Ca2+, but decreased by 2mM Ca2+. Alternatively, the histamine release induced by Hst 8 (10(-6) M) was inhibited by the presence of Ca2+ (0.5 to 2 mM). These results suggest that Hsts have limited usefulness as therapeutic agents due to induction of histamine release from mast cells.     

Effect of interleukin 3 on the differentiation and histamine content of cultured bone marrow mast cells.

Mouse bone marrow hematopoietic stem cells were isolated from mouse femur bone and cultured in RPMI 1640 supplemented medium with 20 units/ml of the purified T-cell lymphokine, interleukin 3 (IL-3), IL-3 was uniquely able to induce the proliferation and differentiation of mature mast cells in vitro. The sparse granulation of the bone marrow-derived mast cells (BMMC) can be seen by day 5, progressing to definable mast cells by day 7, the mast cells appear morphologically mature and comprise a 96% pure population after 14 days of the culture. The monocytes macrophages, eosinophils and neutrophils disappeared by day 9. After 4 weeks of tissue culture, mast cells are fully mature and completely granulated at 98% cell purity. The BMMC are mononuclear, oval or round in shape and appear smaller than rat peritoneal mast cells. BMMC are stable over 3-5 months in conditioned medium. The homogeneous mast cell population possesses membrane receptors and mediators, such as histamine in their metachromatic granules. The histamine content of BMMC in culture between 2 to 4 weeks rose from 1.43 to 1.82 pg/cell. Moreover, the percentage of histamine release caused by 0.1 microM and 1.0 microM ionophore A23187 was 15% and 35%, respectively. By contrast, the histamine releasing activity of 0.01% and 0.001% compound 48/80 were 12 +/- 2% and 59 +/- 7% respectively. The granular density, histamine content and histamine release activity of BMMC are different from that of peritoneal mast cells.