cyclic-gmp and Anaphylaxis

Methylene blue is used primarily in the treatment of patients with methemoglobinemia. Most recently, methylene blue has been used as a treatment for refractory distributive shock from a variety of causes such as sepsis and anaphylaxis. Many studies suggest that the nitric oxide-cyclic guanosine monophosphate (NO-cGMP) pathway plays a significant role in the pathophysiology of distributive shock. There are some experimental and clinical experiences with the use of methylene blue as a selective inhibitor of the NO-cGMP pathway. Methylene blue may play a role in the treatment of distributive shock when standard treatment fails.

Topics: Anaphylaxis; Animals; Antidotes; Cyclic GMP; Drug Resistance; Endothelium, Vascular; Enzyme Inhibitors; Humans; Isoenzymes; Methylene Blue; Models, Biological; Muscle, Smooth, Vascular; Nitric Oxide; Nitric Oxide Synthase; Second Messenger Systems; Sepsis; Shock; Shock, Cardiogenic; Vascular Resistance

Shock can be defined as the failure of the circulatory system to provide necessary cellular nutrients, including oxygen, and to remove metabolic wastes. Although it is now recognized that more than 100 different forms of shock exist, this recognition is more a reflection of the widespread use of the term to describe a variety of disease states. For the purpose of this monograph, we concentrate on various forms of cardiovascular shock, in particular, shock that may be linked to inappropriate vasodilation from overproduction of the endogenous vasodilator, nitric oxide. Some forms of shock have been extensively studied, and convincing evidence exists for the role of nitric oxide. Other disease states have been less well characterized in terms of their association with excess nitric oxide production. Available evidence of a role for nitric oxide is discussed in the hope of stimulating the interest of investigators to explore these areas more thoroughly.

Topics: Anaphylaxis; Animals; Cyclic GMP; Cytokines; Depression, Chemical; Endothelium; Enzyme Inhibitors; Heart; Humans; Nitric Oxide; Nitric Oxide Synthase; Oxygen Consumption; Shock, Cardiogenic; Shock, Septic; Toxins, Biological; Vasodilation

Anaphylactic, anaphylactoid, and related forms of shock are true medical emergencies characterized by bronchial reactivity, airway edema, and hypovolemia. These reactions are attributed to the immunologic and nonimmunologic activation of mast cells and basophils, which results in the acute release of a variety of biochemical mediators that influence vascular tone and permeability. The therapeutic approach to these reactions is guided by the understanding of associated cellular and biochemical changes.

Topics: Anaphylaxis; Animals; Complement Activation; Critical Care; Cyclic AMP; Cyclic GMP; Heart; Hemodynamics; Humans; Immunoglobulin E; Prostaglandins; Resuscitation

Topics: Anaphylaxis; Animals; Anura; Cyclic AMP; Cyclic GMP; Desensitization, Immunologic; Dihydroalprenolol; Dihydroergotoxine; Humans; Hypersensitivity, Immediate; Lymphocytes; Myocardium; Protein Conformation; Receptors, Adrenergic; Receptors, Adrenergic, alpha; Receptors, Adrenergic, beta; Temperature

Topics: Anaphylaxis; Animals; Calcium; Chemotaxis; Chromatography, Gel; Chromatography, Paper; Complement System Proteins; Cyclic AMP; Cyclic GMP; Eosinophils; Esterases; Guinea Pigs; Histamine; Histamine Release; Humans; Hypersensitivity, Immediate; Immunization; Immunoglobulin E; Isoflurophate; Leukocytes; Lung; Lung Compliance; Mast Cells; Peptides; Rats; Serine; Skin; SRS-A

Anaphylactic shock can be defined as an acute syndrome, and it is the most severe clinical manifestation of allergic diseases. Anaphylactoid reactions are similar to anaphylactic events but differ in the pathophysiological mechanism. Nitric oxide (NO) inhibitors during anaphylaxis suggest that NO might decrease the signs and symptoms of anaphylaxis but exacerbate associated vasodilation. Therefore, blocking the effects of NO on vascular smooth muscle by inhibiting the guanylate cyclase (GC) would be a reasonable strategy. This study aimed to investigate the effects of NO/cGMP pathway inhibitors methylene blue (MB), Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), and indigo carmine (IC) in shock induced by compound 48/80 (C48/80) in rats. The effect was assessed by invasive blood pressure measurement. Shock was initiated by C48/80 intravenous bolus injection 5 min before (prophylactic) or after (treatment) the administration of the inhibitors MB (3 mg/kg), L-NAME (1 mg/kg), and IC (3 mg/kg). Of the groups that received drugs as prophylaxis for shock, only the IC group did not present the final systolic blood pressure (SBP) better than the C48/80 group. Regarding shock treatment with the drugs tested, all groups had the final SBP similar to the C48/80group. Altogether, our results suggested that inhibition of GC and NO synthase in NO production pathway was not sufficient to revert hypotension or significantly improve survival.

Topics: Anaphylaxis; Animals; Cyclic GMP; Disease Models, Animal; Enzyme Inhibitors; Indigo Carmine; Male; Methylene Blue; Muscle, Smooth, Vascular; NG-Nitroarginine Methyl Ester; Nitric Oxide; Rats; Rats, Wistar

The purpose of this paper is to propose methylene blue as a lifesaving alternative drug for the treatment of contrast-induced anaphylaxis.. In a cardiovascular catheterization laboratory invasive hemodynamic monitoring was used to document the lifesaving effect of IV bolus injections of 1.5-2 mg/Kg methylene blue solution to treat three patients for anaphylactic shock following radiocontrast injection during coronary angiography. Methylene blue administration was followed by prompt circulatory improvement, leading to hemodynamic stabilization and relief of other anaphylactic symptoms in each case. There were no deaths.. Our findings suggest that methylene blue can be lifesaving in anaphylactic shock, notwithstanding some transitory side effects, such as cardiac rhythm disturbances and chest pain, both of which possibly originate from sudden myocardial perfusion deficits.

Topics: Anaphylaxis; Contrast Media; Cyclic GMP; Enzyme Inhibitors; Humans; Methylene Blue; Nitric Oxide

The effect of a copper amine oxidase (histaminase) purified from the pea seedling as free or immobilized enzyme on the response to specific antigen was studied in isolated hearts from actively sensitized guinea pigs. In vitro challenge with the specific antigen of hearts from actively sensitized animals evokes a positive inotropic and chronotropic effect, a coronary constriction, followed by dilation and an increase in the amount of histamine and nitrites, the oxidation product of nitric oxide, in the perfusates. In the presence of both forms of histaminases, the positive inotropic and chronotropic responses as well as the coronary constriction and the release of histamine were fully blocked. The amount of nitrites, appearing in the perfusates when anaphylaxis is elicited in the presence of both forms of histaminases, is significantly increased, as well as nitric oxide synthase activity and cyclic GMP content in cardiac tissue, while cardiac calcium overload was significantly prevented. These observations demonstrate that the decrease in the anaphylactic release of histamine and the subsequent abatement of the cardiac response to antigen can be accounted for by the inactivation by histaminase of the released histamine and by a stimulation of endogenous nitric oxide production.

Topics: Amine Oxidase (Copper-Containing); Anaphylaxis; Animals; Calcium; Computers; Cyclic GMP; Densitometry; Female; Guinea Pigs; Histamine; Male; Nitric Oxide; Nitric Oxide Synthase; Pisum sativum; Plant Extracts; Time Factors

Clinical and experimental observations prove that heparin-neutralizing doses of protamine increase pulmonary artery pressures and decrease systemic BP. Protamine also increases myocardial oxygen consumption, cardiac output, and heart rate, and decreases systemic vascular resistance. These cardiovascular effects have clinical consequences that have justified studies in this area. Protamine adverse reactions usually have three different categories: systemic hypotension, anaphylactoid reactions, and catastrophic pulmonary vasoconstriction. The precise mechanism that explains protamine-mediated systemic hypotension is unknown. Four experimental protocols performed at Mayo Clinic, Rochester, MN, studied the intrinsic mechanism of protamine vasodilation. The first study reported in vitro systemic and coronary vasodilation after protamine infusion. The second in vitro study suggested that the pulmonary circulation is extensively involved in the protamine-mediated effects on endothelial function. The third study, carried out in anesthetized dogs, reported the methylene blue and nitric oxide synthase blockers neutralization of the protamine vasodilatatory effects. The fourth study suggested that protamine also causes endothelium-dependent vasodilation in heart microvessels and conductance arteries by different mechanisms including hyperpolarization. Reviewing these experimental results and our clinical experience, we suggest methylene blue as a novel approach to prevent and treat hemodynamic complications caused by the use of protamine after cardiopulmonary bypass. In the absence of prospective clinical trials, a growing body of cumulative clinical evidence suggests that methylene blue may be strongly considered as a therapeutic approach in the treatment of distributive shock.

Topics: Anaphylaxis; Animals; Cyclic GMP; Endothelium, Vascular; Hemodynamics; Humans; Methylene Blue; Nitric Oxide; Protamines; Shock

1 Relaxin (RLX) is a multifunctional hormone best known for its role in pregnancy and parturition, that has been also shown to influence coronary perfusion and mast cell activation through the generation of endogenous nitric oxide (NO). In this study we report on the effects of RLX on the biochemical and mechanical changes of ex vivo perfused hearts isolated from ovalbumin-sensitized guinea-pigs induced by challenge with the specific antigen. The possible involvement of NO in the RLX action has been also investigated. 2 A 30-min perfusion with RLX (30 ng ml(-1)) before ovalbumin challenge fully abated the positive chronotropic and inotropic effects evoked by anaphylactic reaction to the antigen. RLX also blunted the short-term coronary constriction following to antigen challenge. Conversely, perfusion with chemically inactivated RLX had no effect. 3 The release of histamine in the perfusate and the accumulation of calcium in heart tissue induced by antigen challenge were significantly decreased by RLX, while the amounts of nitrites in the perfusate were significantly increased, as were NO synthase activity and expression and cGMP levels in heart tissue. 4 These findings indicate that RLX has a protective effect in cardiac anaphylaxis which involves an up-regulation of the NO biosynthetic pathway.

Topics: Anaphylaxis; Animals; Blotting, Western; Calcium; Cell Degranulation; Cyclic GMP; Densitometry; Guinea Pigs; Histamine Release; In Vitro Techniques; Male; Mast Cells; Myocardium; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Ovalbumin; Relaxin; Up-Regulation

1. Haeme oxygenase (HO) is an enzyme mainly localized in the smooth endoplasmic reticulum and involved in haeme degradation and in the generation of carbon monoxide (CO). Here we investigate (1) whether the inducible isoform of HO (HO-1) is expressed in the isolated heart of the guinea-pig and (2) the functional significance of HO-1 on the response to antigen in isolated hearts taken from actively sensitized guinea-pigs. 2. Both the HO-1 expression and activity are consistently increased in hearts from guinea-pigs pretreated with hemin, an HO-1 inducer (4 mg kg(-1) i.p., 18 h before antigen challenge). The administration of the HO-1 inhibitor zinc-protoporphyrin IX (ZnPP-IX, 50 micromol kg(-1), i.p., 6 h before hemin) abolished the increase of both the HO-1 expression and activity. 3. In vitro challenge with the specific antigen of hearts from actively sensitized animals evokes a positive inotropic and chronotropic effect, a coronary constriction followed by dilation and an increase in the amount of histamine in the perfusates. In hearts from hemin-pretreated animals, antigen challenge did not modify the heart rate and the force of contraction; the coronary outflow was significantly increased and a diminution of the release of histamine was observed. The patterns of cardiac anaphylaxis were fully restored in hearts from animals treated with ZnPP-IX 6 h before hemin. 4. In isolated hearts perfused with a Tyrode solution gassed with 100% CO for 5 min and successively reoxygenated, the response to antigen was similar to that observed in hearts from hemin-pretreated animals. 5. Pretreatment with hemin or the exposure to exogenous CO were linked to an increase in cardiac cyclic GMP levels and to a decrease of tissue Ca(2+) levels. 6. The study demonstrates that overexpression of HO-1 inhibits cardiac anaphylaxis through the generation of CO which, in turn, decreases the release of histamine through a cyclic GMP- and Ca(2+)-dependent mechanism.

Topics: Anaphylaxis; Animals; Blotting, Western; Calcium; Carbon Monoxide; Coronary Circulation; Cyclic GMP; Enzyme Inhibitors; Guinea Pigs; Heart; Heart Rate; Heme Oxygenase (Decyclizing); Heme Oxygenase-1; Hemin; Histamine Release; Immunization, Passive; Injections, Intraperitoneal; Male; Myocardial Contraction; Myocardium; Organ Culture Techniques; Perfusion; Protoporphyrins

Cardiac anaphylaxis, an acute ischemic dysfunction comprising coronary vasoconstriction and arrhythmias, is a model of clinically recognized immediate hypersensitivity reactions affecting the heart. Bradykinin, a mediator of hypersensitivity, is also a potent coronary vasodilator, acting via nitric oxide and prostacyclin production. Because ischemia increases bradykinin outflow from the heart, we questioned whether bradykinin might mitigate anaphylactic coronary vasoconstriction. Antigen challenge of hearts isolated from presensitized guinea pigs was associated with an approximately 30% increase in bradykinin overflow. Furthermore, (1) when the half-life of bradykinin was prolonged with the kininase II/angiotensin-converting enzyme inhibitors captopril and enalaprilat, anaphylactic coronary vasoconstriction was attenuated and reversed, and arrhythmias were alleviated; (2) the bradykinin B2-receptor antagonist HOE 140 prevented these effects; and (3) HOE 140 exacerbated both anaphylactic coronary vasoconstriction and arrhythmias. During cardiac anaphylaxis, the coronary overflow of cGMP, a marker of nitric oxide production, and 6-ketoprostaglandin F1 alpha, a stable prostacyclin metabolite, increased two-fold and fourfold, respectively. Because neither enalaprilat nor HOE 140 affected these changes, the enhanced overflow of cGMP and 6-ketoprostaglandin F1 alpha is likely to reflect the actions of other hypersensitivity mediators (eg, histamine and leukotrienes). We postulate that bradykinin plays a protective role in cardiac anaphylaxis by accumulating at the luminal surface of the coronary endothelium and promoting, in an autocrine mode, a B2-receptor-mediated production of nitric oxide and prostacyclin in concentrations sufficient to elicit a paracrine effect on coronary vascular smooth muscle, thus opposing the vasoconstricting effects of other anaphylactic mediators.

Topics: Anaphylaxis; Animals; Arrhythmias, Cardiac; Bradykinin; Captopril; Coronary Vessels; Cyclic GMP; Epoprostenol; Guinea Pigs; Male; Nitric Oxide; Vasodilation

The degree to which pulmonary angiography may contribute to serious complications in patients with pulmonary hypertension has not been clarified and remains a matter of debate. Accordingly, this study was designed (1) to detect the potential release of vasoactive peptides and (2) to investigate the hemodynamic response after administration of a nonionic contrast medium in patients with pulmonary hypertension undergoing pulmonary angiography. Allergy-mediating substances also were measured to monitor for possible anaphylactoid reactions.. Pulmonary digital subtraction angiography was performed in 20 patients with pulmonary hypertension (mean pulmonary arterial pressure more than 20 mm Hg). Iopromide was administered as a total of 100 mL via a 7F catheter inserted from the right femoral vein. The injected volume and duration of injection (15 to 20 mL/sec) were kept constant. Hemodynamic parameters were continuously monitored, including electrocardiogram, heart rate, phasic and mean pulmonary arterial and peripheral arterial pressures. Blood samples were obtained before and after administration of contrast media to assay for the concentration of the following vasoactive peptides using radioimmunoassay techniques: renin, angiotensin-I-converting enzyme, angiotensin II, aldosterone, atrial natriuretic peptide, antidiuretic hormone, cyclic-guanosine monophosphate, and myoglobin, as well as allergy-mediating substances such as tryptase, eosinophil protein X, and eosinophil cationic protein.. Administration of iopromide caused significant increases in atrial natriuretic peptide (from 61.3 +/- 11.8 to 94.0 +/- 16.7) and antidiuretic hormone (from 6.6 +/- 1.9 to 12.3 +/- 3.1), whereas renin significantly decreased (from 3.0 +/- 0.6 to 1.3 +/- 0.5). After administration of contrast media, there were no significant changes in the other measured vasoactive peptides, allergy-mediating substances, and monitored cardiovascular parameters.. Administration of iopromide for pulmonary angiography in patients with pulmonary hypertension resulted in no appreciable hemodynamic alterations associated with the observed changes in atrial natriuretic peptide, antidiuretic hormone, and renin. No allergy-mediated reactions were observed in these patients.

Topics: Adult; Aged; Aldosterone; Anaphylaxis; Angiography, Digital Subtraction; Angiotensin II; Atrial Natriuretic Factor; Blood Pressure; Blood Proteins; Chymases; Contrast Media; Cyclic GMP; Electrocardiography; Eosinophil Granule Proteins; Eosinophil-Derived Neurotoxin; Female; Heart Rate; Humans; Hypertension, Pulmonary; Inflammation Mediators; Iohexol; Lung; Male; Middle Aged; Myoglobin; Peptides; Peptidyl-Dipeptidase A; Renin; Ribonucleases; Serine Endopeptidases; Tryptases; Vasopressins

The effect of several anti-atherogenic drugs (ticlopidine, nicotinic acid and etofibrate) on immune responses and immune complex anaphylaxis has been studied in mice. All the drugs enhanced the activation by concanavalin A, phytohemagglutinin, and lipopolysaccharide of lymphocytes taken from treated animals. Contact hypersensitivity to trinitrochlorobenzene was inhibited by similar treatments with the same drugs, possibly through inhibition of the efferent phase of the reaction. Nicotinic acid produced a slight enhancement of antibody responses to sheep erythrocytes, whereas etofibrate inhibited the response at the highest dose studied. In addition, treatment with these drugs variably protected the mice from anaphylactic shocks induced by immune complexes. Marked protection was also observed using the antiserotoninic, cyproheptadine. These results indicate that drugs used to prevent atherogenic processes modulate different proliferative and effector immunological reactions.

Topics: Anaphylaxis; Animals; Antibody Formation; Antigen-Antibody Complex; Arteriosclerosis; Clofibrate; Clofibric Acid; Cyclic AMP; Cyclic GMP; Female; Hypersensitivity, Delayed; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Niacin; Thiophenes; Ticlopidine

The effects of atropine, 2 mg/kg i.v., on anaphylactic shock were studied in guinea-pigs sensitized to ovalbumin. Atropine only moderately reduced (--31%) the increase in pulmonary resistance observed and slightly prolonged (+26%) the survival time in pretreated animals compared with controls. These effects, however, were no statistically significant. The drug temporarily improved ventilation but had no influence on haematosis. On the other hand, atropine significantly reduced the amount of histamine released (--60%) and of GMPc synthetized in the lung (--21%). The levels of AMPc and prostaglandins E1, E2 and F2 alpha remained comparable to those of control animals. These results suggest that the reflex-induced action of the cholinergic system during anaphylaxis primarily affects large-calibre airways and that the role of acetylcholine in severe reactions is moderate when compared with the direct action of other mediators.

Topics: Airway Resistance; Anaphylaxis; Animals; Atropine; Cyclic AMP; Cyclic GMP; Guinea Pigs; Histamine; Male; Parasympathetic Nervous System; Plethysmography, Whole Body; Prostaglandins; Respiratory Function Tests

Anaphylactic shock was induced with ovalbumin in sensitized rats and the relationship between PGE2 and cyclic nucleotides in lung tissue and plasma histamine during anaphylactic shock was studied. PGE2 level and cyclic AMP/cyclic GMP ratio decreased with this ovalbumin-challenge, and the former reached a minimum value 40 sec after the challenge while the latter reached a minimum value 20 sec later. The plasma histamine level was elevated and reached a maximum value concomitant with the minimum value in the cyclic AMP/cyclic GMP ratio. Dibutyryl cyclic AMP elevated the PGE2 level significantly and inhibited the ovalbumin-induced elevation of plasma histamine, however, this effect was abolished by the administration of indomethacin. PGE2 infusion elevated the cyclic AMP level as well as the cyclic AMP/cyclic GMP ratio, in a time-dependent manner, and inhibited the ovalbumin-induced elevation of plasma histamine during 10 min infusion. There was a significant correlation between the cyclic AMP level and the cyclic AMP/cyclic GMP ratio, both elevated by PGE2 infusion. Thus, anaphylactic elevation of the plasma histamine level results from a decrease in the levels of PGE2 in lung tissue rather than a decrease in the cyclic AMP/cyclic GMP ratio, albeit these decreases being coincident during anaphylactic shock.

Topics: Anaphylaxis; Animals; Cyclic AMP; Cyclic GMP; Histamine; Indomethacin; Lung; Male; Ovalbumin; Prostaglandins E; Rats

Topics: Anaphylaxis; Cell Communication; Cyclic AMP; Cyclic GMP; Diphenhydramine; Histamine; Histamine Release; Humans; Lung; Prostaglandins; Prostaglandins F; Pyrilamine; Time Factors

We studied the effects of Ascaris suum antigen (iv) in 32 natively allergic, anesthetized dogs. After thoracotomy, bilateral cervical vagotomy, and propranolol, samples of peripheral lung frozen in situ were obtained for measurement of cyclic nucleotides and histamine. Following Ascaris, lung histamine decreased 20.4 +/- 3.7% (mean +/- SE), cAMP increased 391 +/- 122%, and cGMP increased 110 +/- 20% with increased plasma histamine and physiological changes of anaphylaxis. No significant changes occurred in 10 control dogs. Release of histamine, reflecting immunological degranulation of mast cells, correlated closely with the physiological effects of anaphylaxis. beta-Adrenergic stimulation with isoproterenol prevented these physiological effects of anaphylaxis in three dogs studied. Furthermore, the level of cAMP induced in lung tissue by beta-adrenergic stimulation in these dogs correlated with the degree of inhibition of immunologically induced histamine release. These results illustrate the suitability of this experimental preparation to study the biochemical and physiological mechanisms of anaphylaxis in vivo.

Topics: Anaphylaxis; Animals; Antigens; Ascaris; Cyclic AMP; Cyclic GMP; Dogs; Histamine; Lung; Lung Compliance

The effect of the antigen-induced, immunoglobulin (Ig)E-dependent release of mediators from human lung tissue was analyzed for coincident changes in the tissue levels of cyclic nucleotides. Simultaneously with the appearance of mediators, lung cyclic guanosine 3',5'-monophosphate (GMP) increased from 0.9+/-0.2 to 12.63+/-4.5 pmol/mg protein and cyclic AMP increased threefold from the initial levels of 5.1+/-1.4 pmol/mg protein. The release of histamine and prostaglandin (PG)F(2alpha), as well as the associated increases in cyclic nucleotides, peaked within 10 min of anaphylaxis. Antagonists of histamine's H-1 receptor prevented anaphylaxis-associated increases in cyclic GMP, whereas H-2 antagonists prevented the cyclic AMP response. Neither of these antagonists influenced the pattern or quantity of histamine or slow-reacting substance of anaphylaxis release. Prevention of PGF(2alpha) synthesis with acetylsalicylic acid failed to influence histamine or slow-reacting substance of anaphylaxis release or the concomitant increases in cyclic nucleotides. Histamine, added exogenously, produced a prompt increase in the cyclic AMP and cyclic GMP levels of human lung. As was seen after anaphylaxis, H-1 anatagonists prevented the cyclic GMP response to histamine, whereas H-2 antagonists prevented the cyclic AMP response.H-1 antagonists prevented 50% of the PGF(2alpha) synthesis accompanying anaphylaxis; H-2 antagonists had no effect. Exogenous histamine induced PGF(2alpha) synthesis; this synthesis was prevented by H-1 but not H-2 antagonists, and was reproduced by 2-methylhistamine (H-1 agonist) but not by dimaprit (H-2 agonist). Arachidonic acid generation of PGF(2alpha) was not influenced by antihistamines. Therefore, histamine interactions with human lung result in the synthesis of both PGF(2alpha) and cyclic GMP in response to H-1 stimulation, and of cyclic AMP through H-2 stimulation.

Topics: Anaphylaxis; Arachidonic Acids; Aspirin; Cyclic AMP; Cyclic GMP; Histamine H1 Antagonists; Histamine H2 Antagonists; Histamine Release; Humans; Immunoglobulin E; In Vitro Techniques; Lung; Prostaglandins F; SRS-A

Topics: Anaphylaxis; Asthma; Cyclic AMP; Cyclic GMP; Desensitization, Immunologic; Epinephrine; Homeostasis; Humans; Lung; Mast Cells; Respiratory Therapy; Vagus Nerve

Topics: Anaphylaxis; Animals; Cyclic AMP; Cyclic GMP; Histamine Release; Humans; Lung; Microsomes; Passive Cutaneous Anaphylaxis

A method is described for the dissociation of guinea pig lung and mesentery into their component cells. The method comprises incubation of the tissues with the enzyme pronase in Ca++- and Mg++-free saline followed by mechanical dispersion and yields on average 3-8% free mast cells. These cells are morphologically intact and viable. They retain an active sensitization or can be passively sensitized. The levels of cAMP and cGMP in the whole cell suspension respond to catecholamine, histamine or cholinergic stimulation in a way similar to that observed in the corresponding undispersed tissue. Although the separation of guinea pig mast cells from other cells proved to be more difficult than the purification of rat mast cells, an inrichment of greater than 50% has been achieved. The mast cells isolated by this method proliferate in vitro and have been cultured successfully for several months.

Topics: Anaphylaxis; Animals; Cells, Cultured; Cyclic AMP; Cyclic GMP; Female; Guinea Pigs; Histamine Release; Immunization, Passive; Lung; Male; Mast Cells; Mesentery; Microscopy, Electron; Organ Specificity; Pronase

A close relationship between increased concentrations of cyclic GMP in human lung tissue and the capacity for acetylcholine to enhance the immunologic secretion of histamine and SRS-A has been found. Acetylcholine (10(-7) to 10(-11) M) produced parallel increases in both cyclic GMP and the immunologic release of mediators; the muscarinic blocking agent atropine prevented both responses. The increase in cyclic GMP in human lung after acetylcholine stimulation was apparent within 30 sec, peaked by 120 sec, and abruptly returned to control levels thereafter. The ability of acetylcholine to enhance the antigen-stimulated secretion of mediators followed the same time-course. PGF2alpha (3.3 X 10(-4) M to 3.3 X 10(-7) M) increased the cyclic GMP content of human lung tissue in a dose-related fashion. Pretreatment of IgE-sensitized lung tissue with acetylsalicylic acid (10 microgram/ml) had no effect on baseline cyclic nucleotide levels, the capacity for antigen to induce mediator release, or the increase in cyclic GMP and facilitation of the immunologic release of mediators produced by acetylcholine.

Topics: Acetylcholine; Anaphylaxis; Antigens; Aspirin; Atropine; Carbachol; Cyclic AMP; Cyclic GMP; Histamine Release; Humans; In Vitro Techniques; Kinetics; Lung; Prostaglandins F; SRS-A

Topics: Anaphylaxis; Animals; Cyclic AMP; Cyclic GMP; Guinea Pigs; Histamine Release; In Vitro Techniques; Kinetics; Lung; Male; Mast Cells; Metiamide; Perfusion; Prostaglandins F

The addition of specific antigen to IgE-sensitized human lung tissue causes the secretion of the mediators histamine and slow-reacting substance of anaphylaxis. The mechanisms by which increased levels of cyclic AMP suppress and increased levels of cyclic GMP enhance this secretory process were studied. Colchicine, an agent which disrupts many secretory reactions by binding to microtubules in their disassembled 6S form, was a relatively ineffective inhibitor of the antigen-induced release of mediators unless lung fragments were incubated at 4 degrees C for 60 min to induce microtubular disassembly. As colchicine appeared to inhibit the immunologic secretion of mediators from human lung tissue most effectively after microtubular disassembly, the capacity of colchicine to modulate the release reaction indicated the state of microtubular assembly; inhibition by colchicine signaled a shift to the colchicine-sensitive 6S subunits whereas failure to inhibit suggested maintenance in the colchicine-resistant polymerized state.Exogenously added 8-Bromo-cyclic GMP prevented low temperature-facilitated colchicine suppression of mediator release suggesting that increased levels of cyclic GMP stabilize polymerized microtubules. Transiently increased cyclic AMP concentrations, either exogenously added as 8-Bromo-cyclic AMP or endogenously produced by isoproterenol, promoted colchicine suppression of mediator release suggesting that microtubular disassembly was produced. Direct measurement of cyclic AMP levels revealed parallel kinetics after isoproterenol stimulation between control and colchicine-treated lung fragments. The requirement for functional microtubules in the release reaction may occur after the antigen IgE-stimulated activation of a serine esterase, energy utilization, and an intracellular calcium requirement. The mechanism by which cyclic nucleotides influence microtubular assembly is postulated to involve the degree of phosphorylation-dephosphorylation of microtubules.

Topics: Anaphylaxis; Colchicine; Cold Temperature; Cyclic AMP; Cyclic GMP; Edetic Acid; Histamine Release; Humans; Isoproterenol; Lung; Microtubules; Nucleotides, Cyclic; SRS-A

The effects of exogenous nucleotides on the histamine hypersensitivity of pharmacologically beta-blocked mice were investigated. Female HLA-SW (ICR) mice, 27-29 gm, were injected intraperitoneally with 20 to 100 mug of propranolol 45 min before intraperitoneal challenge with 1 mg histamine. These animals had a mortality which averaged approximately 80%. At various time intervals before histamine, doses of from 0.5 to 12 mumoles of nucleotides were administered intravenously. Noncyclic nucleotides, adenosine, adenosine 5'-monophosphate (AMP), and guanosine 5'-monophosphate (GMP) showed clear, dose-response protection against histamine death of propranolol-treated mice when they were given 45 to 90 min before histamine. Cyclic AMP showed significant protection only when it was given at a dose of 8 mumoles 45 to 90 min before histamine, and lower or higher doses gave equivocal or no protection. Cyclic GMP WAS Not protective at any dose tested. Propranolol treatment also produced enhanced sensitivity to passive systemic anaphylaxis. Mice were passively sensitized by intraperitoneal injection of mouse anti-egg albumin antibody 6 hr before intravenous challenge with 0.5 mg egg albumin. The mortality from anaphylaxis in the group treated with 20 mug propranolol 45 min before antigen challenge increased to 83%, while that of the group not given propranolol was only 10%. Nucleotides were given intravenously 45 min before antigen challenge. The nucleotides that protected mice from death due to histamine challenge also protected them from death due to systemic anaphylaxis. These protective nucleotides were the same nucleotides that had been reported previously to be protective against Bordetella pertussis-induced hypersensitivity to histamine and anaphylaxis.

Topics: Adenosine; Adenosine Monophosphate; Anaphylaxis; Animals; Cyclic AMP; Cyclic GMP; Dose-Response Relationship, Drug; Drug Hypersensitivity; Female; Guanine Nucleotides; Histamine; Lethal Dose 50; Mice; Mice, Inbred ICR; Nucleotides; Propranolol

Topics: Anaphylaxis; Antigens; Carbachol; Chemotaxis; Cyclic AMP; Cyclic GMP; Eosinophils; Esterases; Glycolysis; Histamine Release; Humans; In Vitro Techniques; Isoflurophate; Lung; Receptors, Adrenergic; Receptors, Cholinergic; SRS-A

Topics: Anaphylaxis; Animals; Bucladesine; Cyclic AMP; Cyclic GMP; Depression, Chemical; Dose-Response Relationship, Drug; Female; Fluorometry; Histamine Release; Mast Cells; Ovalbumin; Rats

Topics: Analysis of Variance; Anaphylaxis; Animals; Antigen-Antibody Reactions; Benzyl Compounds; Carbon Radioisotopes; Cyclic AMP; Cyclic GMP; Ethylenediamines; Guinea Pigs; Histamine; Histamine H1 Antagonists; Humans; Imidazoles; In Vitro Techniques; Iodine Radioisotopes; Lung; Ovalbumin; Pyridines; Rabbits; Radioimmunoassay; Thiourea; Tritium

Topics: Adenosine Diphosphate; Adenosine Monophosphate; Adenosine Triphosphate; Anaphylaxis; Animals; Bordetella pertussis; Bucladesine; Corticosterone; Cyclic AMP; Cyclic GMP; Deoxyribonucleotides; Female; Histamine; Hypersensitivity, Immediate; Hypoglycemia; Inosine Nucleotides; Mice; Nucleotides; Prostaglandins; Spleen; Thymine Nucleotides; Thymus Gland; Time Factors

The capacity to extract slow-reacting substance of anaphylaxis (SRS-A) from human lung tissue or cells after immunologic activation, together with the measurement of SRS-A in both the extract and the surrounding fluid, permits study of total SRS-A generation. That the material extracted is SRS-A was established by both differential bioassay and purification. SRS-A accumulation was entirely intracellular after limited IgE-dependent direct or reversed anaphylactic activation. Intracellular accumulation also generally preceded release, with generation of SRS-A continuing well beyond a plateau in the cellular SRS-A level and the release of preformed mediators. The quantity of SRS-A generated after immunologic activation was modulated by the introduction of exogenous cyclic nucleotides, revealing a site of cyclic nucleotide action distinct from that on mediator release. The capacity to determine not only the release of preformed mediators but also the generation of a newly formed mediator, the sum of SRS-A in cells and supernate, adds an additional dimension to the analysis of the cellular events of immediate hypersensitivity.

Topics: Anaphylaxis; Animals; Antigens; Biological Assay; Bucladesine; Cyclic AMP; Cyclic GMP; Guinea Pigs; Humans; Immunization; Immunoglobulin E; Immunoglobulin G; Lung; Rats; SRS-A; Tissue Extracts

The immunologic release of histamine and slow reacting substance of anaphylaxis (SRS-A) from human lung tissue can be enhanced by stimulation with either alpha adrenergic agents (phenylephrine or norepinephrine in the presence of propranolol) or cholinergic agents (acetylcholine or Carbachol). The finding that atropine prevents cholinergic but not comparable alpha adrenergic enhancement is consistent with the view that cholinergic and alpha adrenergic agonists interact with separate receptor sites on the target cells involved in the immunologic release of chemical mediators. The consistent qualitative relationship between the antigen-induced release of mediators and the level of cyclic adenosine monophosphate (cyclic AMP) as measured by the isolation of (14)C-labeled cyclic AMP after incorporation of adenine-(14)C into the tissues or by the cyclic AMP binding protein assay suggests that changes in the level of this cyclic nucleotide mediate adrenergic modulation of the release of histamine and SRS-A. The addition of 8-bromo-cyclic guanosine monophosphate (cyclic GMP) produces an enhancement of the immunologic release of mediators while dibutyryl cyclic AMP is inhibitory. As cholinergic-induced enhancement was not associated with a measurable change in the levels of cyclic AMP, the possibility is suggested that cyclic GMP may be the intracellular mediator of cholinergic-induced enhancement of the immunologic release of histamine and SRS-A.

Topics: Acetylcholine; Adenine; Anaphylaxis; Atropine; Autacoids; Carbachol; Carbon Isotopes; Cyclic AMP; Cyclic GMP; Drug Antagonism; Histamine Release; Humans; In Vitro Techniques; Lipid Metabolism; Lung; Norepinephrine; Phenylephrine; Propranolol; Receptors, Adrenergic; Receptors, Cholinergic; Receptors, Drug; Stimulation, Chemical