Biomedical subjects

G J Gleich

Publications and source records attributed to G J Gleich.

At least 289 records · Page 16Linked to original sources

PubMed1987

The injurious effect of eosinophil peroxidase, hydrogen peroxide, and halides on pneumocytes in vitro.

Recent data suggest that eosinophils may cause lung injury. To determine if the eosinophil peroxidase (EPO)-hydrogen peroxide (H2O2)-halide system could mediate this injury, we added human EPO, H2O2 (or glucose and glucose oxidase as a continuous source of H2O2), and various halides to monolayers of 51Cr-labeled human A549 and rat type II pneumocytes. Cell lysis was measured as soluble 51Cr release. In initial experiments, EPO in solution did not induce lysis under these conditions. Therefore, in subsequent experiments, pneumocytes were preincubated with EPO for 15 minutes, washed to remove unbound enzyme, and then glucose, glucose oxidase, and the halides were added. EPO alone was not injurious, nor was the addition of glucose and glucose oxidase in the absence of EPO. In contrast, the combined addition of EPO, glucose, glucose oxidase, and chloride produced marked target-cell lysis. This effect was time and EPO dose dependent and was enhanced by the addition of iodide. Catalase and azide substantially inhibited the lysis produced by the EPO-H2O2-halide system, suggesting that EPO-catalyzed products of halide oxidation mediated this form of injury. Finally, the addition of eosinophil major basic protein at 10(-5) mol/L to EPO-coated pneumocytes incubated with glucose, glucose oxidase, and halides failed to enhance or inhibit lysis. We hypothesize that the EPO-H2O2-halide system may injure the lung in asthma and eosinophilic pulmonary syndromes.

Animals↗

PubMed1987

Extracellular deposition of eosinophil granule major basic protein in pressure urticaria.

Utilizing affinity chromatography-purified antibody to the eosinophil granule major basic protein and formalin-fixed paraffin embedded tissue, we investigated the localization of major basic protein by immunofluorescence in twenty-four skin biopsy specimens from ten patients with pressure urticaria. Fourteen of twenty-four biopsy specimens were obtained from spontaneously occurring urticarial lesions of 4 to 48 hours' duration, and ten of the twenty-four specimens were from dermographometer-induced lesions that had been present from 40 minutes to 24 hours. Twenty-one of twenty-four biopsy specimens showed extracellular fluorescence of eosinophil granule major basic protein within the dermis. The extent and intensity of extracellular staining were not related to the presence or degree of tissue eosinophilia. Serial section controls from each block were stained with protein A purified rabbit IgG and were negative. Previous immunofluorescence studies have demonstrated deposition of major basic protein in lesions of chronic idiopathic urticaria, episodic angioedema, and facial edema. Major basic protein causes release of histamine from human basophils and induces wheal-and-flare reactions on intradermal injection. The present observations add further evidence to support a role for eosinophil mediators, particularly major basic protein, in the pathogenesis of cutaneous disease characterized by edema.

Adult↗

PubMed1987

Increases of plasma eosinophil major basic protein levels late in pregnancy predict onset of labor.

Major basic protein (MBP), the core of the eosinophil granule, is a potent toxin for parasites and mammalian cells; it also causes histamine release from mast cells and basophils. Plasma levels of MBP are increased in all pregnant women, and MBP has been localized by immunofluorescence to placental X cells and placental-site giant cells. Analyses of plasma MBP levels during pregnancy showed a striking correlation between increases late in the third trimester and spontaneous onset of labor. In one case with premature labor, the plasma MBP level also was increased coincidentally with labor. In contrast, in cases of prolonged gestation in which labor was induced, there were no late increases in MBP levels. Also, uterine contraction during spontaneous labor or oxytocin-induced labor did not cause elevations of plasma MBP. Measurement of plasma MBP levels in pregnancy may be useful for the prediction of labor and for the development of new means of intervention for premature labor and prolonged gestation.

Blood Proteins↗

PubMed1987

Bronchoalveolar cell profiles of asthmatic and nonasthmatic subjects.

Asthma is associated with increased airway responsiveness to pharmacologic agents such as methacholine. Increases in airway responsiveness after exposure to allergen or ozone are associated with increased inflammatory cells in bronchoalveolar lavage both in human and in animal studies. We studied the total and differential cell counts in 10 stable atopic asthmatics who had airway hyperresponsiveness but no clinical features of airway inflammation and 10 nonasthmatic subjects, using a conventional 100-ml lavage and a 20-ml washing. Metachromatic cell numbers and eosinophils were increased in both the lavage (p less than 0.01 for metachromatic cells; p = 0.05 for eosinophils) and washing (p less than 0.025 for metachromatic cells and p = 0.03 for eosinophils) compared with those in nonasthmatics. In asthmatics, metachromatic cell numbers in the lavage and washing, and total cell count and, to a lesser extent, eosinophils in lavage were significantly correlated with measurements of airway responsiveness. Major basic protein in lavage and blood did not differ between asthmatics and nonasthmatics. The washing, although it revealed a different cell profile from the lavage in both normal subjects and asthmatics, did not show differences between asthmatics and nonasthmatics undetected by the lavage specimen. We conclude that there is evidence of cellular inflammation in the airway of stable asthmatics and that small volume washings do not add to the information concerning the cell profile of asthmatics and nonasthmatics provided by conventional lavage.

Asthma↗

PubMed1987

The effect of purified human eosinophil major basic protein on mammalian ciliary activity.

Eosinophil granulocyte infiltration in a variety of lung disorders may directly damage local tissue by release of granule contents. A principal constituent of eosinophil granules is the major basic protein (MBP). Previous light and electron microscopic observations have indicated that guinea pig and human MBP produce detachment of tracheal epithelial cells and cessation of ciliary activity. To quantitate the damage, selected regions of the epithelial surface of rabbit tracheal explants were videotaped before and after treatment with human MBP. Tapes were analyzed for ciliary beat frequency and the extent of zones along the epithelial surface displaying ciliary activity. The MBP at 0.1 mg/ml reduced beat frequency and significantly reduced the measured zones of ciliary activity. Also, MBP at 0.7 mg/ml significantly reduced beat frequency and the measured zone of ciliary activity on the epithelial surface. Beat frequency was lowered by 27% within 10 min, with only 1% further decrease by 60 min. The zones of ciliary activity on the epithelium were continuously decreased throughout the 60 min to 29% of the initially active zone. To examine whether MBP was capable of direct inhibition of ciliary activity, isolated porcine tracheal ciliary axonemes, the structural organelles of individual cilia, were treated with human MBP. Concentrations above 67 micrograms/ml of MBP were completely inhibitory to reactivated isolated axonemes, 67 micrograms/ml stopped activity within 10 min, and 27 micrograms/ml stopped activity within 15 min. Pretreatment of isolated axonemes with increasing concentrations of MBP resulted in decreasing ATPase activity. These effects were not attributable to pH alteration.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗

PubMed1987

Comparative toxicity of purified human eosinophil granule proteins for newborn larvae of Trichinella spiralis.

Eosinophils have been implicated in both in vivo and in vitro destruction of helminths. One approach toward elucidating the role of the eosinophil in parasite killing has been to test the toxicity of purified eosinophil granule proteins for parasites in vitro. Previously, major basic protein (MBP) and eosinophil cationic protein (ECP) were shown to be toxic for schistosomules of Schistosoma mansoni, while eosinophil-derived neurotoxin (EDN) was only marginally so. We tested the toxicity of MBP, ECP, and EDN over a range of concentrations (0.006-5 X 10(-4) M) for newborn larvae of Trichinella spiralis. Our observations confirm previous reports of toxicity of mildly reduced and alkylated (R & A) MBP. At concentrations of 5 X 10(-5) M and above, R & A MBP killed 75% or more of the larvae within the first hour of culture. ECP was an effective toxin for these larvae after 3 hr of culture, and by 12 hr, dose-related toxicity was evident. After 24 hr, 100% of the larvae were killed at 5 X 10(-5) M ECP. EDN was much less toxic; after 12 hr, 90% of the larvae survived at concentrations of 1 X 10(4) M, while 5 X 10(-4) M EDN killed all the larvae. At the optimal toxic concentrations of 5 X 10(-5) M ECP and 5 X 10(-4) M EDN, kinetics of killing by these 2 proteins were essentially the same. Thus, on a molecular basis, both MBP and ECP appear to be potent helminthotoxins whereas EDN is much less so.

Animals↗

PubMed1986

Ribonuclease activity associated with human eosinophil-derived neurotoxin and eosinophil cationic protein.

The eosinophil granule contains a series of basic proteins, including major basic protein, eosinophil peroxidase, eosinophil-derived neurotoxin (EDN), and eosinophil cationic protein (ECP). Both EDN and ECP are neurotoxins and helminthotoxins. Comparison of the partial N-terminal amino acid sequences of EDN and ECP showed 67% identity; surprisingly, they also showed structural homology to pancreatic ribonuclease (RNase). Therefore, we determined whether EDN and ECP possess RNase enzymatic activity. By spectrophotometric assay of acid soluble nucleotides formed from yeast RNA, purified EDN showed RNase activity similar to bovine pancreatic RNase, whereas ECP was 50 to 100 times less active. The RNase activity associated with ECP was not significantly inhibited after exposure of ECP to polyclonal or monoclonal antibody to EDN. These results indicate that EDN and ECP both possess RNase activity, the RNase activity of EDN and ECP is specific, and EDN and ECP have maintained not only structural but also functional homology to pancreatic RNase.

Blood Proteins↗

PubMed1986

Differences between basophils and mast cells: failure to detect Charcot-Leyden crystal protein (lysophospholipase) and eosinophil granule major basic protein in human mast cells.

Human eosinophils contain several distinctive proteins including eosinophil granule MBP and the membrane-associated CLC protein (lysophospholipase). Human basophils also contain these proteins, indicating biochemical similarities between eosinophils and basophils. To determine whether MBP or CLC protein is present in connective tissue mast cells, we studied human lung and cutaneous mast cells by immunofluorescence by utilizing specific antibodies to CLC and MBP. Cytocentrifuge slides of enriched lung mast cells and mast cells in sections of formalin-fixed, paraffin-embedded cutaneous tissue from urticaria pigmentosa lesions were stained for CLC and MBP. Neither pulmonary nor cutaneous mast cells stained for CLC protein or MBP. In contrast, lung and cutaneous eosinophils in the same preparations showed bright staining for both proteins. The failure to find CLC protein and MBP in mast cells provides additional evidence of dissimilarity between mast cells and basophils, and an immunochemical means to distinguish between them.

Basophils↗

PubMed1986

Chronic eosinophilic pneumonia. Evidence for eosinophil degranulation and release of major basic protein.

A patient with chronic eosinophilic pneumonia associated with ulcerative colitis and asthma is described. Exacerbations and remissions of these disorders paralleled each other. Pleural fluid and lung tissue contained high levels of eosinophil granule major basic protein, a substance with toxic effects on cells and tissues. The results indicate that eosinophils degranulate in the lung in eosinophilic pneumonia and point to the possibility that granule constituents damage lung tissue in the course of the disease.

Adult↗

PubMed1986

Effects of glucocorticoids on eosinophil colony growth.

The in vivo and in vitro effects of glucocorticoids on eosinophilopoiesis were examined with soft agar cultures of bone marrow and peripheral blood cells. Prednisone, 10 mg four times daily for three days, administered to normal volunteers, caused a significant drop in circulating eosinophil numbers (p less than 0.005) but did not decrease eosinophil colony numbers in cultures of bone marrow or peripheral blood. A patient with peripheral blood eosinophilia demonstrated a larger percentage decrease in mean eosinophil colony numbers (50%) after prednisone than did any normal volunteer. Incubation of bone marrow cells for 1 hour with either high concentrations of hydrocortisone, up to 3.3 X 10(-4) mol/L, or with postinfusion plasma from volunteers administered intravenous hydrocortisone, plasma levels to 1.6 X 10(-5) mol/L, did not decrease the numbers of eosinophil colonies. At a concentration of 3.3 X 10(-6) mol/L of hydrocortisone, there was a slight but statistically significant stimulation of eosinophil colony numbers. In contrast, incubation with high concentrations of dexamethasone, 3.3 X 10(-4) mol/L or 3.3 X 10(-6) mol/L, significantly reduced eosinophil colony numbers. Prednisone caused a significant reduction in plasma levels of Charcot-Leyden crystal protein but not of eosinophil granule major basic protein. The results indicate that soft agar assay of eosinophil colony growth by blood or bone marrow cells cannot be used to model the in vivo eosinopenic effect of glucocorticoids, levels of dexamethasone in excess of levels commonly administered in clinical practice are required to inhibit eosinophilopoiesis in vitro, and patients with peripheral blood eosinophilia may be more susceptible to the eosinopenic effects of glucocorticoids than normal subjects.

Blood Proteins↗

PubMed1986

Microbiologic analyses of nasal polyp tissue.

Nasal polyps from 40 patients were cultured within 2 1/2 hours after surgical removal to determine whether microorganisms were present. The first 20 polyps were cultured for aerobic and anaerobic bacteria, viruses, fungi, mycoplasmas, and mycobacteria. Of these 20 polyps, eight were sterile by all tests, one grew Cryptococcus albidus, one grew Sporobolomyces, one had large numbers of Peptostreptococcus micros and Propionibacterium acnes, greater than 10(6) colony-forming units per gram (cfu/gm), and nine had aerobic bacteria including 10 different species at levels less than 10(5) cfu/gm. The second 20 polyps were cultured for aerobic bacteria only; 11 polyps were positive. Overall, 14 of 26 polyps from patients with asthma and two of 14 polyps from patients without asthma were positive for aerobic bacteria at levels greater than 10(3) cfu/gm (p less than 0.05). Multiple aerobic bacterial species tended to occur in polyps from patients with asthma (11 of 26) more frequently than in those from patients without asthma (one of 14) (p less than 0.01). There was a highly significant positive correlation between tissue neutrophilia and bacterial count (r = +0.9; p less than 0.001). The results indicate that patients with asthma have a significantly higher number and a tendency to a greater variety of aerobic bacteria in nasal polyp tissue than patients without asthma and that the number of infiltrating neutrophils is directly related to the number of bacteria.

Adolescent↗

PubMed1986

Detection of antibodies to proteases used in laundry detergents by the radioallergosorbent test.

Two proteases, Esperase and Alcalase, derived from Bacillus licheniformis and B. subtilis, respectively, are used in laundry products. In testing for the prevalence of IgE antibodies to these enzymes in sera among 300 laundry product workers, we experienced two problems in the establishment of a reliable RAST for these antigens. The first problem was the propensity of the allergen, Esperase, to undergo autolysis, suggesting that solid-phase Esperase might also lose reactivity through degradation. Treatment of Esperase with phenylmethylsulfonyl fluoride stabilized the enzyme and permitted the synthesis of a stable solid-phase antigen. The second problem was the finding that sera reactive with Esperase in the RAST were also reactive with Savinase, an enzyme from B. licheniformis to which the workers were not exposed. Immunochemical analyses of the three enzymes with specific rabbit antisera by gel diffusion and by two-site immunoradiometric assay demonstrated that they were not cross contaminated to any appreciable extent. RAST inhibition demonstrated that solid-phase Esperase possessed unique allergenic determinants in that the reactivity of IgE antibodies was inhibited by low concentrations of Esperase and only by very high concentrations of Alcalase and Savinase. In contrast, the reactivity of solid-phase Alcalase was occasionally inhibited equally well by Esperase and Alcalase. Most strikingly, the reaction of IgE antibodies with solid-phase Savinase was always inhibited by comparable quantities of Esperase, Alcalase, and Savinase. Thus, the establishment of the RAST for these proteases appears to require the use of phenylmethylsulfonyl fluoride to retard autolysis, and the results must be interpreted with caution because IgE antibodies in certain sera demonstrate cross-reactivity with Alcalase and Savinase.

Antibodies↗

PubMed1986

The eosinophil and the pathophysiology of asthma.

Eosinophilia of lung and blood associated with injury to the mucociliary escalator and excessive shedding of bronchial epithelium are hallmarks of both allergic and nonallergic asthma. In vitro, the eosinophil granule major basic protein (MBP) is toxic to helminths and to mammalian cells, including human respiratory epithelium. The MBP-mediated damage to the respiratory epithelium consists of desquamation and frank destruction of ciliated cells. Increased sputum MBP concentration is a good marker for asthma, and patients treated for acute asthma have high levels of MBP in their sputa, which decrease after treatment. Peak sputum MBP levels approximate concentrations toxic to respiratory epithelium in vitro. In the lungs of patients who had died of asthma, MBP has been localized outside of the eosinophil in association with damage to the epithelium. Overall, these and other findings suggest the hypothesis that the eosinophil mediates damage to the respiratory epithelium and is the prime effector cell in the pathophysiology of asthma.

Animals↗

PubMed1986

Variability of IgE protein measurement in cell-culture supernatants: results from a multicenter collaborative study.

The sensitivity, specificity, and precision of immunoassays for quantitation of IgE in cell-culture supernatants were tested in a multicenter trial involving 22 laboratories. Fourteen coded test samples included cell-culture medium alone, culture medium with varying concentrations of polyclonal or myeloma IgE, and medium from unstimulated or pokeweed mitogen (PWM)-stimulated peripheral blood mononuclear cell (PBMC)-culture supernatants. Two laboratories reported measurable IgE in non-IgE-containing control samples. Although the IgE content of a single 0.50 ng/ml polyclonal IgE sample should have been measured easily by the claims of all laboratories, only 13 laboratories measured IgE in this sample in 22 of the 48 assays. Most laboratories could measure both polyclonal and myeloma IgE at 5.0 ng/ml; however, the IgE determinations for the myeloma proteins were nearer the predicted value. Although 15 laboratories found measurable IgE in the PWM-stimulated PBMC-culture supernatants from a single nonatopic donor, the levels did not differ significantly from that measured in unstimulated PBMC-culture supernatants. Three laboratories reported considerably higher IgE levels in the PWM-stimulated PBMC-culture supernatants than in other PBMC-culture supernatants. Only 13 laboratories could quantitate IgE in each of coded duplicate samples containing 0.50 ng/ml polyclonal IgE. These findings indicate a wide variability in the sensitivity, specificity, and precision of assays used to quantitate low levels of IgE protein. Investigators should be encouraged to make greater use of existing national and international reference materials in the standardization and performance of their IgE immunoassays.

Double-Blind Method↗

PubMed1986

The eosinophilic leukocyte: structure and function.

The evidence reviewed here indicates that the eosinophil has the ability to kill many species of helminths and likely does so during worm infection. This toxic ability appears to be regulated by several other cells including mast cells, monocytes, and T lymphocytes. Eosinophils kill helminths through their ability to generate potent oxidants and through their content of cationic proteins, which likely achieve high concentrations at points of granule deposition. Eosinophils also participate in inflammation in human disease especially asthma, skin diseases, and heart disease. Though present concepts hold that the mast cell is the cornerstone of the allergic inflammatory response (450), the findings that eosinophils bind IgE and are activated by antigen-IgE complexes and that the eosinophil can elaborate many inflammatory mediators raise the possibility that the eosinophil might also be involved in the initiation of inflammatory responses. Finally, an eosinophil-related protein appears to play an undefined role in human reproduction.

Animals↗

PubMed1986

The eosinophil and cutaneous edema.

Although eosinophils are readily identified in skin tissue, their role in cutaneous disease has been obscure. Recent studies have elucidated the structure, content, and several activities of the eosinophil. The eosinophil is a potent parasite-killer cell and probably mediates damage to respiratory epithelium in bronchial asthma. We review information showing an association between cutaneous edema and eosinophil degranulation in tissue. These studies show that eosinophils release and deposit toxic granule proteins extensively in the skin despite the existence of few intact eosinophils in tissue. The evidence suggests that the eosinophil functions not only as a parasite-killer cell but also as a proinflammatory cell that may be pathophysiologically related to the development of cutaneous edema.

Adult↗

PubMed1986

Biochemical and functional similarities between human eosinophil-derived neurotoxin and eosinophil cationic protein: homology with ribonuclease.

Eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) were isolated from lysates of human eosinophil granules by gel filtration and ion exchange chromatography on heparin-Sepharose. Radioimmunoassay, using monoclonal antibodies, of fractions from the heparin-Sepharose chromatography showed one peak of EDN activity and two peaks of ECP activity (termed ECP-1 and ECP-2). EDN, ECP-1, and ECP-2 each exhibited heterogeneity in charge and molecular weight when analyzed by two-dimensional nonequilibrium pH gradient electrophoresis and NaDodSO4/PAGE. Digestion of EDN with endoglycosidase F (endo F) decreased its molecular weight and charge heterogeneity. Thus, END likely contains a single complex oligosaccharide. Endo F digestion of ECP-1 and ECP-2 decreased the molecular weight of both polypeptides, indicating that both likely contain at least one complex oligosaccharide. Amino acid sequence analyses showed that ECP-1 and ECP-2 are identical from residue 1 through residue 59 and that the sequences of EDN and ECP are highly homologous (37 of 55 residues identical). Both EDN and ECP NH2-terminal sequences showed significant homology to RNase, especially in regions of the RNase molecule involved in ligand binding. EDN, ECP-1, and ECP-2 had neurotoxic activity, causing the Gordon phenomenon at doses down to 0.15 micrograms when injected into the cisterna magna; the proteins were comparable in their activities. These results indicate that EDN and ECP are related proteins and suggest that they derived from genes associated with the RNase family.

Amino Acid Sequence↗

PubMed1986

Immunochemical quantitation of an airborne proteolytic enzyme, Esperase, in a consumer products factory.

Occupational exposure of dry bleach factory workers to proteolytic enzymes poses a potential health hazard to susceptible workers. A sensitive immunochemical technique has been developed for quantitating exposure of factory workers to Esperase by sampling airborne particles on a fiberglass filter sheet followed by elution and assay for enzyme in a two-site immunoradiometric assay. By this method, 0.05 to 0.1 ng of enzyme (activity 50 KNPU/g) can be quantitated. With this sensitivity, as little as 2 to 4 ng/m3 airborne Esperase could be detected by personal monitors (flow rate of 0.002 m3/min; sampling time of 8 hr) and 0.05-0.1 ng/m3 of airborne Esperase with high volume samplers (flow rate of 1.13 m3/min;sampling time of 8 hr). There were significant day-to-day and site-to-site variations in the airborne Esperase content in the factory. Significant variations were also noted in the quantities of Esperase/m3 in the air inhaled by the workers in the same area on the same day. With this sensitive immunochemical technique and the use of personal monitors, the actual exposure of the worker as he moves about the factory could be measured.

Air Pollutants, Occupational↗