Antibody determination against Aspergillus fumigatus by means of the enzyme-linked immunosorbent assay. I. Liberation of polystyrene binding components during cultivation.

The liberation of polystyrene binding antigenic components (PBC) of Aspergillus fumigatus involved in ELISA IgG measurements was studied during cultivation in the culture filtrates. PBC were rapidly liberated into the culture medium during the logarithmic phase of growth (phase I), were in part degraded or reconsumed during the stationary phase (phase II) and again secreted (other PBC) during the lytic phase (phase III). ELISA IgG titers and numbers of precipitating components are not always related with one another, may show an inverse relationship; maximal ELISA titers are reached during phase I when precipitating components are limited.

Reduced adenylate cyclase responsiveness to histamine in lymphocyte membranes of allergic asthmatic patients after allergen challenge.

The adenylate cyclase response to histamine was measured in lymphocyte membranes of allergic asthmatic patients and healthy control subjects, just before and 24 h after inhalation challenge with house dust mite allergen. In the non-acute phase before allergen challenge, the histamine response in the membranes of the patients was not significantly different from that in membranes of the control subjects. After the house dust mite-induced asthmatic reaction, however, the adenylate cyclase response to histamine in the membranes of the patients was significantly reduced by 47%, whereas allergen inhalation had no effect on the histamine response of the control membranes.

Standardization of allergenic extracts of Aspergillus fumigatus. Liberation of IgE-binding components during cultivation.

During cultivation of Aspergillus fumigatus a rapid liberation of IgE-binding components was found reaching maximum values during the logarithmic phase of growth (phase I). After a fall in IgE-binding titers during phase II, appearance of additional IgE-binding components was noted during the period of lysis of the microorganism (phase III). These latter allergenic components are different from the phase I IgE-binding components, as was shown by crossed-inhibition studies.

A volumetric-aerobiologic study of seasonal fungus prevalence inside and outside dwellings of asthmatic patients living in northeast Netherlands.

Air sampling with an Andersen sampler was carried out over a period of 21 mo, in 28 houses of asthmatic patients with an allergy for moulds. Inside, the livingroom, bedroom, and kitchen were studied, and an additional outside sample was always taken. Indoors, Penicillium showed the highest concentration, especially in the autumn and winter months, while Aspergillus and yeasts were also found frequently.

Immunologic studies in bronchoalveolar fluid in a patient with allergic bronchopulmonary aspergillosis.

Bronchoalveolar lavage was performed in a patient during an acute stage of allergic bronchopulmonary aspergillosis (ABPA) and repeated 17 mo later during a remission stage and cessation of corticosteroid therapy. Measurement of protein concentrations (albumin, IgG, IgA, and IgM) by laser nephelometry and crossed immunoelectrophoresis indicates that transudation of proteins occurs from the circulation into the lung compartment during the acute stage of ABPA. Furthermore, comparing total Ig concentrations and titers of ELISA IgG, IgA, and IgM antibodies against Aspergillus fumigatus with the corresponding values of serum measurements indicates a preferential local production of IgA antibodies and IgM to a lesser extent.

Immunologic observations in sera of a patient with allergic bronchopulmonary aspergillosis by means of the enzyme-linked immunosorbent assay.

Titers of IgG, IgA, and IgM antibodies against Aspergillus fumigatus were measured by ELISA in sera of a patient with allergic bronchopulmonary aspergillosis during an acute stage, remission, and exacerbation of the disease. Total IgE, specific IgE against A. fumigatus, and number of precipitation lines were also measured.

Urinary N tau-methylhistamine during early and late allergen-induced bronchial-obstructive reactions.

Urinary N tau-methylhistamine, a unique histamine metabolite, was determined in 29 patients with bronchial asthma before and after inhalation of a control solution on 1 day and before and after allergen inhalation on another day. Twenty-eight patients had an early bronchial-obstructive reaction, defined by a decrease in FEV1 of 20% or more within 1 hr after allergen inhalation. A late bronchial-obstructive reaction, 3 to 8 hr after allergen inhalation, was found in 16 patients.

Measurement of plasma histamine by stable isotope dilution gas chromatography-mass spectrometry: methodology and normal values.

A new method for the determination of histamine by stable isotope dilution mass fragmentography is described. The method is specific, sensitive, and accurate, resulting in a within-day coefficient of variation of 4.1% and a day-to-day variation of 7.

Measurement of specific IgE and IgG antibodies against Aspergillus fumigatus antigen in patient sera by use of enzyme immunoassays: influence of different procedures of antigen immobilization.

IgG and IgE antibody titers against Aspergillus fumigatus were measured in patient sera by enzyme immunoassays by use of antigens or allergens immobilized to different carriers. Specific-IgG antibodies were measured by a double antibody-layer enzyme immunoassay; specific IgE was determined by Phadezym -RAST (Pharmacia Diagnostics). In both cases antigens and allergens were immobilized in two ways: first by covalent binding to CNBr-activated paper discs and second by spontaneous binding to polystyrene surface of microtiter plates.

Effect of primary renal disease in patients with renovascular insufficiency.

The effect of laboratory evidence of renal parenchymal abnormality on the results of renal revascularization in 83 patients with renovascular hypertension was determined. Primary renal disease (PRD) was defined as an abnormal urinalysis (proteinuria, hematuria, or casts) in the absence of urinary infection, or decreased renal function (elevated serum creatinine level greater than 1.5 mg/dl and/or decreased creatinine clearance).

The allergenic and antigenic properties of spore extracts of Aspergillus fumigatus: a comparative study of spore extracts with mycelium and culture filtrate extracts.

The antigenic properties of conidial preparations of Aspergillus fumigatus were compared with mycelial and culture filtrate extracts by use of sera of patients with aspergillosis and hyperimmune rabbit serum. Biologic activities were tested by intracutaneous tests. The immunologic parameters used were: precipitating properties in double-diffusion, IgG binding by ELISA, IgE binding by enzyme inhibition assay, and by enzyme allergosorbent test.

Environmental aerobiological studies in allergic bronchopulmonary aspergillosis.

An aerobiological pilot study was undertaken to measure airborne concentrations of Aspergillus spores in and around the dwellings of two patients with allergic bronchopulmonary aspergillosis (ABPA). The aim of the investigation was 1) to find a specific source of Aspergillus antigen, and 2) to find out whether disease activity, or exacerbation, paralleled Aspergillus spore prevalence. In the environment of the first patient heavy spore contaminated air was found in a cow-shed, and the adjacent hayloft and scullery.

Dynamics of the lymphocyte beta-adrenoceptor system in patients with allergic bronchial asthma.

It has been proposed that reduced beta-adrenergic responsiveness plays an important role in the increased airway reactivity of asthmatic patients. This hypothesis has been supported by studies showing reduced beta-adrenergic responsiveness in lymphocytes of asthmatic patients, predominantly during the occurrence of active and severe symptoms. Little is known about the mechanism underlying this relationship and its clinical relevance with respect to bronchial hyperreactivity.

Comparison of antibody measurements against Aspergillus fumigatus by means of double-diffusion and enzyme-linked immunosorbent assay (ELISA).

IgG antibody titers against Aspergillus fumigatus from different sera were measured by means of a standardized ELISA and compared with precipitates measured by double diffusion (D.D.).

Plasma histamine concentrations and complement activation during house dust mite-provoked bronchial obstructive reactions.

Lung function and histamine levels in peripheral venous plasma samples were followed after challenge with house dust mite allergen. In eight patients the mean maximal histamine concentration showed a significant rise during the early obstructive reaction when compared with the concentrations at similar time intervals after inhalation of a control solution. In nine other patients treatment with disodium cromoglycate (DSCG) caused a significant fall in plasma histamine concentrations indicating that histamine liberation plays a role during the early obstructive reaction and was released from the lung compartment.

A single centrifugation step method for the simultaneous separation of different leukocytes with special reference to basophilic leukocytes.

A simple method of separating mononuclear leukocytes, basophils, neutrophils and an eosinophil fraction from human blood is described, using discontinuous Percoll gradient centrifugation. Mononuclear cells were found in the upper layer (band 1) at a density of less than or equal to 1.072.

Characterization of (-)-[3H]dihydroalprenolol binding to intact and broken cell preparations of human peripheral blood lymphocytes.

In this study we compared characteristics of (-)-[3H]dihydroalprenolol ([3H]DHA) binding sites in crude membrane preparations of human peripheral blood lymphocytes with those of intact, viable cells. A valid determination of specific beta-adrenergic receptor binding in both preparations was obtained by defining non-specific [3H]DHA binding with 10(-6) M l- or dl-propranolol or 10(-3) M l-isoproterenol. Higher concentrations of propranolol were used in prior reports on lymphocyte membranes.

Bronchial obstructive reactions after inhalation with endotoxin and precipitinogens of Haemophilus influenzae in patients with chronic non-specific lung disease.

Bronchial obstructive reactions occur after inhalation of Haemophilus influenzae in the absence of proteolytic activity in patients with CNLSD and a lowered histamine threshold in whom specific precipitins are demonstrated. It may be presumed, that the direct bronchial reaction is caused by its content of endotoxin, whereas the late bronchial reaction is the result of a toxic action and a type III allergic reaction. When proteolytic activity is present it lowers the histamine threshold and may influence the endotoxin effect in this way.

The beta-adrenergic system and allergic bronchial asthma: changes in lymphocyte beta-adrenergic receptor number and adenylate cyclase activity after an allergen-induced asthmatic attack.

Beta-adrenergic receptor characteristics and adenylate cyclase responses to different stimuli were measured in lymphocyte membrane preparations of six normal control subjects and five allergic asthmatic patients with mild airways disease and increased bronchial reactivity to histamine and acetylcholine. The determinations were performed just before and 24 hr after inhalation challenge with house-dust mite antigen to investigate the influence of an allergen-induced asthmatic attack on the beta-adrenergic receptor system. Before the house-dust mite challenge, the lymphocyte membranes of the patients showed a normal receptor density, dissociation constant for (-)3H-dihydroalprenolol, and adenylate cyclase response to isoproterenol, guanyl-5'-yl-imidodiphosphate, (GppNHp) and NaF.