C. A. Hirshman

Comparison of Histamine Release in Human Skin Mast Cells Induced by Morphine, Fentanyl, and Oxymorphone

Human leukocyte and skin mast cell preparations were incubated with morphine sulfate in concentrations ranging from 1.5 × 10−5. M to 4.5 × 10−5 M. Skin mast cells also were incubated with oxymorphone and fentanyl in the same concentrations. Human leukocytes did not release histamine in response to any concentration of morphine. In skin mast cells, histamine release by morphine first was detected at 1.5 × 10−4 M. Histamine release further increased at 5.0 × 10−4 M with no incremental increase at higher concentrations. Oxymorphone and fentanyl failed to release histamine at any concentration. Histamine release by morphine required calcium but was not influenced by changes in the 1–4 nM range. Skin mast cell preparations were pretreated for 30 min in naloxone 5 × 10−4 M and then morphine 5 × 10−4 M was added for 30 min without removing naloxone. Naloxone neither released histamine nor inhibited morphine-induced histamine release. The release of histamine by morphine but not equimolar concentrations of fentanyl and oxymorphone indicates that histamine release by narcotics is not a nonspecific effect of high drug concentration. The failure of naloxone to inhibit morphine-induced histamine release suggests that histamine release by morphine is not dependent on opiate receptor binding or activation. These results indicate that this human mast cell preparation will be useful in further understanding the mechanism of histamine release induced by morphine and other agents.

Pruritic dermatitis in asthmatic basenji-greyhound dogs: A model for human atopic dermatitis

A recurrent, nonseasonal pruritic dermatitis manifested as lichenified plaques and as inflammatory nodules and papules in each of fourteen basenji-greyhound (B-G) crossbreed dogs, all of which demonstrated both nonspecific and Ascaris-specific airway hyperreactivity, with changes in pulmonary mechanics of the same order as those seen in symptomatic human asthma. Other features analogous to human atopic disease included blunted cyclic adenosine monophosphate (cAMP) responsiveness to beta adrenergic agents and in vivo release of histamine and slow-reacting substance of anaphylaxis in response to antigen aerosol challenge. The dermatitis appears to be a manifestation of the atopic state in these dogs and may provide an animal model for the study of human atopic dermatitis.

Thiobarbiturate-induced Histamine Release in Human Skin Mast Cells

Human skin mast cell preparations were incubated with thiopental, thiamylal, methohexital, and pentobarbital in concentrations ranging from 10−5 M to 10−3 M. Both thiopental- and thiamylal-induced dose-related histamine release, with thiamylal having a significantly greater effect than thiopental (P < 0.05). In contrast, incubation of skin mast cell preparations with the same concentrations of pentobarbital and methohexital failed to increase histamine release above spontaneous levels at any concentration. The release of histamine by thiopental and thiamylal was not accompanied by the leakage of lactic dehydrogenase (LDH). Although a demonstration of histamine release in vitro is not proof that clinical symptoms are causally related to histamine release in vivo, methohexital may be preferred as the induction agent in patients showing extreme sensitivity to histamine (asthmatics) or increased histamine releasability (atopics).

Functional differences between human cutaneous mast cells and basophils: a comparison of morphine-induced histamine release

Intravenous administration of morphine sulfate often produces urticarial and hypotensive reactions associated with elevations in plasma histamine. The source of this histamine and mechanisms controlling its release are poorly understood. Previous studies of morphine-induced histamine release compared human leukocytes to rat peritoneal mast cells. The effects of morphine on human cutaneous mast cells has not been examined. We studiedin vitro histamine release from human basophils and human skin preparations containing cutaneous mast cells to evaluate their relative, contribution to the pharmacologic effects of morphine.Human skin mast cell preparations showed dosedependent histamine release over a morphine concentration range of 1.5×10−5 to 4.5×10−3M, with peak release occurring at 5×10−4M, with peak release occurring at 5×10−4M. Clinically, morphine sulfate is usually injected as a 1.5×10−2M solution. Histamine release was calcium dependent and equivalent to that obtained with 3 and 10 mM strontium. Morphologic examination revealed degranulation and exocytosis occurring in morphine-stimulated tissue but not in specimens exposed to buffer alone. Lactate dehydrogenase levels did not increase following morphine incubation, thus supporting a noncytolytic mechanism of histamine release.Basophils, in contrast, showed no significant histamine release from exposure to morphine up to 10−2M. Concanavalin A, as a positive control in these same preparations, produced a mean histamine release of 21.0%.Our studies indicate distinct functional differences between human skin mast cell and human blood basophil responses to morphine sulfate. We conclude that the cutaneous and systemic reactions to morphine sulfate probably result from the release of histamine from mast cells rather than from basophils.

Anaphylactic reaction during anaesthesia associated with positive intradermal skin test to fentanyl

A 29 year-old female patient suffered vascular collapse which became apparent immediately after general anaesthesia. Resuscitation was prolonged and difficult, and complicated by the need for reoperation. Based on the time history, fentanyl was suspected as the causative agent. Fentanyl allergy was confirmed by skin testing one month later. The case is discussed, and the possible reasons for the delay in appearance of symptoms and signs are considered.RésuméUn patient a démontré un choc cardiovasculaire ayant apparu immédiatement après l’anesthésie générale. La réanimation était prolongée et difficile et compliquée par une réopération. En se basant sur l’histoire chronologique, le fentanyl a été suspecté comme étant l’agent causal. L’allergie au fentanyl a été confirmée par des tests cutanés un mois plus tard. Le cas est discuté et les raisons possibles pour le délai dans l’apparition des symptômes et des signes est discuté.

Airway constrictor effects of leukotriene D4 in dogs with hyperreactive airways

Leukotriene D4 (5 micrograms/ml) aerosol constricts airways of dogs with nonspecific airway hyperreactivity but not of mongrel dogs which lack nonspecific airway hyperreactivity. RL increased 200 +/- 25% and Cdyn decreased to 77 +/- 5% of the prechallenge value. LTD4 (10 micrograms/ml) produced no further increase. Atropine (0.2 mg/kg) prevented the increase in RL and decrease in Cdyn, suggesting that part of the effect of LTD4 on airways is neurally mediated.

Elevated mononuclear leukocyte phosphodiesterase in allergic dogs with and without airway hyperresponsiveness

To investigate if mononuclear leukocyte beta-adrenergic hyporesponsiveness of Basenji greyhound (BG) dogs is associated with atopy or nonspecific airway hyperresponsiveness, we examined the relationship between mononuclear leukocyte cAMP phosphodiesterase levels, airway responsiveness to methacholine, and intradermal allergen responses in 17 BG dogs, five unrelated purebred Basenjis, and five greyhounds. BG dogs were hyperresponsive to aerosols of methacholine compared to Basenjis and greyhounds. Both BG dogs and Basenjis were allergic and had increased leukocyte cAMP phosphodiesterase activity compared to greyhounds. We concluded that the leukocyte abnormality is not associated with airway hyperresponsiveness. The leukocyte abnormality is either associated with the allergic state, with some hereditary trait that BG dogs acquired from the Basenji ancestry, or the leukocyte abnormality is necessary but not sufficient for the development of airway hyperresponsiveness.

Enhanced bronchoalveolar lavage mast cells histamine releasability in allergic dogs with and without airway hyperresponsiveness.

To determine if mast cells from the airway lumen of Basenji-greyhound (BG) dogs differ functionally from mast cells of control dogs, we compared spontaneous release and A 23187-induced and C5-induced histamine release from bronchoalveolar lavage (BAL) fluid of 14 BG and five allergic and five nonallergic control dogs. No dog received antigen, agonist, or therapeutic aerosols for 4 weeks before BAL. The fluid recovered was centrifuged, and the number of mast cells was quantitated. Aliquots containing equal numbers of mast cells were incubated with A 23187 or C5 for 30 minutes, and histamine release was measured by an automated fluorometric method. Spontaneous release, A 23187-induced release, C5-induced release, and total histamine content per mast cell were calculated. The total amount of histamine per mast cell was not significantly different in BGs and allergic and nonallergic control dogs. Mast cells obtained by BAL released histamine to A 23187 and C5 in a dose-related manner. Spontaneous histamine release and A 23187-induced histamine release was significantly greater in BGs and allergic control dogs compared to nonallergic control dogs. C5-induced histamine release was significantly greater in BGs than in allergic as well as nonallergic control dogs. These data suggest that BAL mast cell histamine releasability must be defined with respect to each stimulus and that mast cells obtained from BAL from control dogs differ with respect to histamine releasability from mast cells of allergic dogs with and without airway hyperresponsiveness. This study suggests that mast cells obtained from BAL can be used to study mast cell function in the control and the allergic state.

Inhibition of d-Turbocurarine-induced Histamine Release by Halothane

To determine whether anesthetics modify mediator release, the authors measured the amount of histamine released by d-tubocurarine (dTC) in human foreskin preparations in the presence of high (2.0%) and low (0.5%) halothane concentrations and nitrous oxide (10%). Freshly excised human foreskins were divided into four matched pieces. Two matched pieces were aerated with oxygen, and the other two with an oxygen-anesthetic gas mixture. One chamber of each served as a control, while the other was stimulated with 3 X 10(-5) M d-tubocurarine for 30 min. Supernatant histamine concentrations were measured by automated fluorometry. Percent histamine release was determined by dividing the experimentally released histamine concentration by the total histamine released after the tissue was sonicated and boiled. Neither halothane nor N2O alone altered spontaneous histamine release. Histamine release by d-tubocurarine was significantly reduced by 2% halothane compared to d-tubocurarine alone (2.8% +/- 0.9 vs. 13.9% +/- 3.7, mean + SEM) (P less than 0.05) in the in vitro preparation. Histamine release was reduced in the preparations pretreated with 0.5% halothane group, but this was not statistically significant (P greater than 0.05) when compared to d-tubocurarine alone. N2O (10%) did not reduce d-tubocurarine-induced histamine release. The authors conclude that halothane, in clinically used concentrations, significantly impairs histamine release from human neonatal foreskin preparations.

The effect of calcium on cardiac anaphylaxis in guinea-pig Langendorff heart preparations

This study was designed to determine the effects of different calcium concentrations on the perfused isolated guinea-pig heart preparation subjected to cardiac anaphylaxis. Following challenge both physiological and biochemical effects were determined on hearts from guinea-pigs previously sensitized to ovalbumin. Perfusion media containing either 1, 2.54 or 5 mM of calcium was used.In comparison to nonsensitized controls challenged to ovalbumin, challenged sensitized hearts (CSH) perfused with 1 mM Ca2+ showed an initial increase indF/dt, a prolonged rise in H.R. and depressed coronary flow. Raising the calcium concentration to either 2.54 or 5 mM in CSH preparations resulted in a marked increase in the release of lactate dehydrogenase (LDH) into the coronary effluent and depressed coronary flow. Perfusing CSH preparations with increasingly higher calcium concentrations more often produced severe tachyarrhythmias and fibrillation. The highest level of histamine released into the coronary effluent occurred immediately following challenge and then declined exponentially over the next 20 min. Both challenge and the administration of histamine induced an immediate but transient increase in H.R., a rise indF/dt, and LDH release. The infusion of histamine produced an increase in coronary flow, but on porcine tubular coronary arterial segments only a direct constricting effect was obtained. The prior administration of cimetidine (10−5M) attenuated the rise in LDH anddF/dt in CSH and nonsensitized preparations infused with histamine (3 μg). However, although cimetidine did not affect the decreased coronary flow in CSH preparations, it initially attenuated the rise in coronary flow in preparations infused with histamine.These results suggest that calcium enhances the liklihood of tachyarrhythmias in cardiac anaphylaxis. The release of LDH in histamine-infused preparations and those CSH preparations perfused with 2.54 and 5 mM calcium-containing medias also suggests the possibility that calcium enhances the damaging effects on the myocardial cell in cardiac anaphylaxis.