Automated histamine analysis for in vitro allergy testing. II. Correlation of skin test results with in vitro whole blood histamine release in 82 patients.

The recently developed sensitive, automated histamine assay system was applied for in vitro allergy testing. The simplified method for histamine release from whole heparinized blood was used. Aliquots of blood and allergen were incubated for one hour at 37 degrees C, and each supernatant was then analyzed for histamine release. Nine common pollen and environmental allergens were used at three 10-fold dilutions for in vitro testing with the use of 20 ml of blood. Intradermal skin tests were correlated with the whole blood histamine release in 82 patients who had received no immunotherapy. A scoring system for the histamine results was developed to take into consideration the results with multiple allergen concentrations. When the skin test was strongly positive (greater than or equal to 3 + at 100 protein nitrogen units [PNU]/ml), the whole blood histamine release was positive in 89% of the tests. In contrast, when the skin test was negative ( less than 1 + at 100 PNU/ml), the histamine release was also negative in 99.8% of the cases. When the skin test was 1 +, the histamine release from whole blood was positive in 6% of the tests; and when the skin test was 2+, the whole blood results were positive in 32%. The accuracy, precision, and sensitivity of the automated histamine assay allow its application for the clinical study of allergic patients.     

Competent cellular immunity in allergic rhinitis patients with elevated IgE.

Parameters of cell-mediated immunity (CMI) were studied in 17 allergic rhinitis patients selected for markedly elevated total serum IgE levels (greater than 300 IU/ml) and 14 normal controls. Mean serum IgE levels were 1,421 IU/ml and 101 IU/ml in the allergic and control groups, respectively (p less than 0.001). There were no significant differences between the allergic patients and the normal controls in delayed cutaneous hypersensitivity, in mitogen and antigen lymphocyte transformation in heterologous or autologous plasma, or in percentage of sheep erythrocyte rosettes. The allergic patient group had a significantly higher percentage of sheep erythrocyte-antibody-complement rosettes (p less than 0.05). Markedly elevated total serum IgE levels in allergic rhinitis patients were not associated with any detectable impairment of CMI.     

The direct demonstration of histamine release in allergic reactions in the skin using a skin chamber technique.

We have adapted a skin chamber technique to permit sampling of fluid at the skin window sites of pollen antigen-induced allergic reactions. Low background levels of histamines are formed in control chambers, whereas significantly increased (p less than 0.01) amounts are found within 30 min following ragweed application in sensitized subjects.     

Human lung tissue and anaphylaxis. I. The role of cyclic GMP as a modulator of the immunologically induced secretory process.

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.     

Diagnostic tests in ragweed-allergic asthma. A comparison of direct skin tests, leukocyte histamine release, and quantitative bronchial challenge

Thirty-six patients with a history of seasonal ragweed asthma were tested. Direct intradermal skin tests, leukocyte histamine release, and quantitative inhalation bronchial challenge correlated significantly. Data reported suggest that quantitative skin tests are as diagnostically valuable as quantitative inhalation bronchial challenge or leukocyte histamine release. Data also suggest that lung and skin mast cells and circulating basophils respond as a single population of cells to ragweed antigens.     

Enhancement of antigen-induced leukocyte histamine release by a mononuclear cell--derived factor.

Supernatant fluids of phytohemagglutinin-stimulated mononuclear cells from ragweed-sensitive patients significantly enhanced the release of histamine from antigen-triggered leukocytes of ragweed-sensitive as well as control individuals. Supernatants of mononuclear cells from control individuals did not reveal this enhancing effect, nor was it found with the use of supernatants of unstimulated mononuclear cells of ragweed-sensitive patients or culture media with PHA alone. Supernatant fluids of phytohemagglutinin-stimulated mononuclear cells of patients sensitive to trees and grass also revealed this enhancing effect. The factor(s) responsible for the enhancement of antigen-induced histamine is heat labile and has a molecular weight of less than 10,000 daltons. The mechanism and site of action of the enhancing factor could involve initiating and/or modulating steps of the leukocyte histamine release reaction. This factor, presumably a lymphokine or a monokine, may constitute a regulating link between cell-mediated immunity and histamine-mediated hypersensitivity reactions in allergic patients.     

Polistes wasp hypersensitivity: diagnosis by venom-induced release of histamine in vitro.

Polistes wasps cause a majority of Hymenoptera-induced anaphylactic reactions in Texas. Using the in vitro release of histamine from basophils of patients allergic to Polistes stings, we have studied the cross-reactivity of venoms from three species of Polistes wasps as well as the cross-reactivity among Polistes, honeybee, and Vespula maculifrons (yellow jacket) venoms. Venom collected by an extrusion technique from Pollistes exclamans, Pollistes apachus, and Pollistes carolina caused release of histamine in seven Polistes-sensitive individuals. The dose-response curves from all three Polistes species were quite similar, suggesting extensive cross-reactivity among these species. None of these patients showed significant release of histamine from leukocytes exposed to yellow jacket or honeybee venom. We conclude that a source of Polistes venom is available for further study and possibly for therapy. It appears that any of three local common species of Polistes wasps could be used. Our studies confirmed earlier reports that Hymenoptera sensitivity if often genus-specific.     

Anaphylaxis following psyllium ingestion

Psyllium is a hydrophilic agent found in many bulk laxative preparations. We report the occurrence of an anaphylactic reaction in a patient after ingestion of a psyllium-containing laxative. IgE mediation of the reaction was suggested by a positive immediate skin test to psyllium, positive passive transfer skin test, lack of skin response during passive transfer with heat treated serum, and an elevated IgE (RAST) to psyllium seed.     

Plasma histamine changes during provoked bronchospasm in asthmatic patients

Seven patients with bronchial asthma underwent bronchial inhalation challenge with aerosolized allergen extracts and methacholine. Simultaneously, venous blood samples were collected and histamine was measured. Each patient was challenged on successive days with an allergen extract to which he had no skin-sensitizing antibody (skin test-negative allergen), followed by methacholine and skin test-positive allergen. Bronchospasm was not induced by inhalation of skin test-negative allergens but was observed in all patients after methacholine and in the majority of patients after skin test-positive allergens. No changes in plasma histamine were detected after challenges with methacholine and skin test-negative allergens. After challenge with skin test-positive allergens, significant rises in plasma histamine were detected in 5 of 7 patients. Plasma histamine was elevated within the first 5 min after inhalation of aerosolized allergen, and elevations persisted as long as 30 min. These studies showing that histamine increases significantly in the plasma during allergen-induced asthma in man suggest that histamine should be considered as at least one of the mediators of bronchospasm in allergic asthma. Bronchospasm induced by the cholinergic drug methacholine, unlike allergen-induced bronchospasm, is not associated with changes in plasma histamine.     

Effects of infused histamine on asthmatic and normal subjects: comparison of skin test responses

The effect of histamine infused intravenously at sequentially increasing concentrations (0.05, 0.1, 0.25, 0.5, and 1 μ/kg/min) on the wheat responses to intradermal histamine and compound 48/80 in eight normal and five asthmatic subjects and to allergen skin tests in five asthmatic subjects was measured. These measurements were repeated following pretreatment with the H-1 antagonist hydroxyzine or the H-2 antagonist cimetidine, either alone or in combination. Histamine infused in progressively increasing concentrations had no effect on histamine, compound 48/80, or allergen skin tests either before or after H-1 or H-2 antihistamine treatment. No significant differene was found in the concentration of histamine or compound 48/80 required to elicit a 10-mm wheat in normal or asthmatic patients. Pretreatment with the H-2 antagonist alone had no effect on histamine or compound 48/80 skin tests in either group. However, the H-1 antagonist significantly reduced the wheat response to histamine (p < 0.05 normal; p < 0.025 asthmatics) and compound 48/80 (p < 0.05 normal; p < 0.025 asthmatics) in both groups. The combination of H-1 and H-2 histamine antagonists was not significantly different from the H-1 antagonist alone. Antigen skin testing was suppressed 82% by the hydroxyzine alone; no significant suppression was induced by cimetidine alone, and the combination of hydroxyzine plus cimetidine was only slightly more effective than hydroxyzine alone. The results indicate that blockade of histamine H-2 receptors with cimetidine has little or no additive effect on H-1 antagonist-suppressed skin test responses to histamine, compound , or antigen. Furthermore, the capacity of histamine to suppress histamine release in vitro from basophils was not demonstrated in vivo assessing skin mast cell responses. This observation combined with earlier studies on the human lung mast cell, which also failed to demonstrate that histamine had an inhibitory action, suggests that the human mast cell may not respond to histamine like the basophil and that this discrepancy may represent a fundamental difference in the cell types.     

Assessment of tissue fluid histamine levels in patients with urticaria

Tissue fluid histamine in lesions and normal skin sites of patients with various forms of physically induced urticaria and chronic idiopathic urticaria was assessed utilizing suction-induced blisters. Histamine levels were elevated over challenged sites in cold urticaria, solar urticaria, and delayed pressure urticaria. Histamine levels were elevated in both challenged and “control” sites in patients with immediate-pressure urticaria and local heat urticaria in whom the apparatus provoked lesions. In 5 of 13 patients with chronic idiopathic urticaria, the blister fluid histamine levels were elevated in lesions but not in normal-appearing skin, while in 7 patients the blister fluid histamine levels were elevated in lesions as well as in normal-appearing skin. Although the pathogenesis of chronic idiopathic urticaria is unknown, a clear abnormality related to skin histamine has been identified.     

Effect of histamine and methacholine on nasal airway resistance in atopic and nonatopic subjects: Comparison with bronchial challenge and skin test responses

Serial nasal, intracutaneous, or bronchial challenges were carried out with solutions containing 2- or 3-fold increments in histamine (H) or methacholine (Meth) concentration until nasal airway resistance (NAR) increased by more than 100%, a large intracutaneous reaction was elicited, or FEV₁ decreased by 20% or more. Thirty nonatopic and 48 asymptomatic atopic subjects were studied, the latter group divided into rhinitic patients with and without asthma. Several types of data analysis demonstrated there was no significant difference in the nasal or cutaneous effects of H or Meth between the atopic and nonatopic groups. Comparable results were obtained in a subgroup of 39 subjects (13 normal, 13 atopic, and 13 atopic with asthma) who underwent all six test sequences (i.e., nasal, cutaneous, and bronchial with both drugs). As expected, the asthmatics showed significantly increased bronchial reactivity to both agents. In comparison with Meth, H had a much greater effect on the nasal mucosa and skin than on the bronchi. It is concluded that, contrary to bronchial responses, but in accord with cutaneous reactivity, the nasal responses of nonatopic subjects, atopic persons with allergic rhinitis alone, and subjects with both allergic rhinitis and asthma show no intergroup differences on testing with H or Meth.     

Effect of a selective beta 2 adrenergic agonist and theophylline on skin test reactivity and cardiovascular parameters.

Fenoterol, a selective beta 2-adrenergic agent, and aminophylline in the "therapeutic range" were compared with placebo for their inhibitory effect on skin test reactivity to allergens and histamine. Cardiovascular parameters were also assessed. A new, inexpensive micrometer adaptor to a tuberculin syringe was used to deliver allergens and histamine more accurately. No inhibition of skin test reactivity to antigens or histamine was found after a loading dose or after 1 wk of round-the-clock therapy with these bronchodilators. Although there was increased heart rate after 1 and 2 hr with fenoterol, there was no patient preference for one bronchodilator over the other. The results of this study point out some of the difficulties in trying to extrapolate in vitro findings to in vivo correlates since neither fenoterol nor therapeutic doses of theophylline interfere with immediate skin test reactivity.     

IgE antipolymyxin B antibody formation in a T cell-depleted bone marrow transplant patient

The production of IgE-class antibody specific for polymyxin B is documented in an 18-year-old white female acute myelocytic leukemic patient in relapse. The patient was rendered T cell-deficient by total body X-irradiation and antihuman thymocyte globulin for the purpose of bone marrow transplantation. Thereafter, symptoms of nasal congestion, rhinorrhea, and perinasal urtication produced by topical application of a polymyxin solution were noted. Reaginic activity mediated by an IgE antibody against polymyxin is documented by Prausnitz-Küstner-type passive transfer reactions and by an indirect hemagglutination technique developed for these studies. The occurrence of type I hypersensitivity to this topical antibiotic is rare. It is speculated that pharmaceuticals normally having a low sensitizing potential might demonstrate increased reaginic immunogenicity in a spontaneously or iatrogenically T cell-depleted patient.     

Automated histamine analysis for in vitro allergy testing : I. A method utilizing allergen-induced histamine release from whole blood

A sensitive, automated, histamine assay system has been developed and applied for in vitro allergy testing. Nine common pollen and environmental allergens were used at three log dilutions for in vivo studies utilizing small volumes of blood (15-20 ml). The clinical evaluation was correlated with the results of the histamine release. two different procedures were utilized. The first is the commonly used histamine release from washed leukocytes. There was excellent correlation between the clinical evaluation and the results of histamine release from washed leukocytes in 17 different individuals. The second and simpler method utilized whole heparinized blood which might better reflect the immunologic reaction which occurs in vivo. Aliquots of blood and allergen were incubated for 1 hr at 37 degrees C and each supernatant was then analyzed for histamine release. There was excellent correlation between the two tests in 29 patients tested by both the whole blood and washed leukocyte methods. There was also good correlation between the clinical evaluation of the patients amd the intro tests. The precision, accuracy, and sensitivity of the automated histamine assay make feasible its routine application in the clinical study of allergic patients.     

A comparison of methacholine and histamine inhalations in asthmatics.

Bronchial provocation tests using aerosolized serially diluted histamine and methacholine were given to nearly 200 asthmatics. Results were usually reproducible for a given patient and corticosteroids did not influence the procedures. Those patients who could tolerate high doses of methacholine were statistically the least severe asthmatics as measured by their discharge dose of corticosteroids. Reaginmediated asthmatics could not tolerate the higher doses of histamine. These tests help delineate subgroups of asthmatics and may have clinical usefulness since, when combined with other data, they differentiate pathogenetic mechanisms in some patients and suggest therapeutic approaches in others.     

Complement-mediated release of histamine from human basophils. III. Possible regulatory role of microtubules and microfilaments.

The release of histamine by normal human leukocytes (basophils) following in vitro challenge with activated complement (zymosan-treated serum) was previously reported. In this study, the effects of various pharmacologic agents on this release mechanism were compared with allergen-induced release of histamine. Colchicine and vinblastine antagonize the polymerization of tubulin to form microtubules, and both agents inhibited complement-and allergen-triggered release of histamine from basophils. Finally, treatment with cytochalasin B, a fungal product known to interfere with microfilament formatin, resulted in enhanced release of histamine from complement-treated basophils but no significant change in the percentage of histamine released from allergen-treated basophils. These findings suggest that microtubules and/or microfilaments are involved in complement-induced secretion of histamine by human basophils.     

A controlled study of the effect of corticosteroids on immediate skin test reactivity

Limited uncontrolled studies in the past have yielded conflicting results as to the ability of corticosteroids to suppress the immediate wheal-and-flare test. This double-blind controlled study, using 15 atopic volunteers, demonstrated that a one-week course of steroids exerted no significant effect, compared to placebo, on reactivity to ragweed wheal size and the threshold dilution measurements. Reactivity to compound 48–80 and histamine was also unaffected. In vivo effects of the steroid in these subjects was demonstrated by significant eosinopenia.     

Evaluation of the adverse effects of long-term hyposensitization.

This study was undertaken to determine if long-term hyposensitization causes late sequelae, particularly those reflecting aberrant immunologic responses. Atopic individuals receiving five or more years of hyposensitization with allergenic extracts showed no increased autoimmune, collagen vascular, or lymphoproliferative disease. In addition, chronic hyposensitization did not have adverse effects on immunologic reactivity as assessed by a number of immune parameters. Particularly noteworthy was the absence of immune complexes in the serum of patients undergoing long-term hyposensitization. This study represents the first systematic investigation of potential adverse effects of long-term hyposensitization.