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PubMed1999

The effect of corticosteroid therapy on blood eosinophils and eosinophilic cationic protein in patients with acute and chronic asthma.

There is evidence that eosinophils are involved in inflammation in asthma, a correlation having been observed between blood eosinophil (B-EOS) count and pulmonary function. It has been suggested that eosinophils, and its product, eosinophil cationic protein (ECP), can serve as markers of disease activity. This paper examines this hypothesis. B-EOS count, serum ECP level, and peak expiratory flow (PEF) were estimated in two groups of asthmatics and controls at three visits in 4 weeks. The mean B-EOS count in acute and stable asthmatic groups was higher than in controls at presentation; the difference was statistically significant (p<0.02). Similarly, mean ECP was higher in the two groups than in controls, but with no statistically significant difference. The B-EOS count and serum ECP level within the groups fell between week 0 and week 4 because of treatment. There was positive correlation between ECP and PEF and also between B-EOS and ECP and PEF. The findings reveal that blood eosinophils reflect some degree of activity in asthmatic patients in the acute and chronic state.

Acute Disease↗

PubMed1987

Degranulated eosinophils, eosinophil granule basic proteins and humoral factors in Nigerians with endomyocardial fibrosis.

The frequency of eosinophilia, degranulated eosinophils, and raised serum levels of eosinophil granule basic proteins was determined in ten Nigerians with EMF and fifteen normal controls matched for age and sex, and from the same environment and social background as the patients. Sera from the patients and control subjects were also examined for auto-antibodies, immune complexes, filaria antibodies and immunoglobulins. A mild eosinophilia was observed in four of the patients. The mean direct eosinophil count in the patients (0.407 +/- 0.14 X 10(9)/l) was, however, not significantly different from that of the control subjects (0.399 +/- 0.07 x 10(9)/l). Degranulated eosinophils were absent in the blood and bone marrow aspirates of the patients, and peripheral blood of the controls. It was also found that the mean concentration of eosinophil granule basic proteins (ECP/EPX) in the patients (0.278 +/- 0.1 micrograms/ml) was not significantly different from the mean value for the control subjects (0.371 +/- 0.1 micrograms/ml, P greater than 0.5). Serological studies using Brugia pahangi antigens failed to demonstrate filaria antibodies in the patients and the control subjects. Microfilaria were also absent in their peripheral blood films. Using the indirect immunofluorescent technique, anti-heart antibodies were not demonstrated in the sera of the EMF patients and control subjects. Five (50%) of the patients and four (30.7%) of the normal controls had immune complexes levels far in excess of the upper limit of normal. However, the mean concentration of immune complexes in the patients (16.35%) was not significantly different from the value for control subjects (12.13%, P greater than 0.5).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1984

Surface proteins of the human eosinophil. II. Effects of Schistosoma mansoni larvae on eosinophil surface proteins.

Incubation of eosinophils with schistosoma of Schistosoma mansoni in vitro induces changes in the eosinophil membrane. Cell surface proteins were blocked with unlabelled iodide and then, after incubation with schistosomula, newly accessible proteins were detected by lactoperoxidase catalysed iodination with radioactive iodide. Normal turnover restores several proteins to the eosinophil surface, but the schistosomula specifically induced the appearance of a protein of mol. wt 18K. This protein has previously been detected when eosinophils interact with antibody coated agar layers. Soluble factors released by schistosomula induced the appearance of enhanced levels of 16 and 18K proteins. It has previously been shown that proteins involved in binding of eosinophils to IgG coated antigen coupled cellulose have mol. wts of 16K and 18K.

Antigen-Antibody Complex↗

PubMed1993

[Review: recent studies on a human eosinophilic leukemia cell line, EoL as an experimental model of eosinophils].

Recently, a human eosinophilic leukemia cell line, EoL was established from an eosinophilic leukemia patient. EoL-1 cells have the cytohistologic features of myeloblasts under normal culture conditions, and they can be induced to differentiate into eosinophilic granule-containing cells but not into other lineage cells under several culture conditions and are therefore considered to be committed precursors of eosinophils. Furthermore, EoL-1 cells can also be induced to differentiate functionally to show PAF-induced Ca2+ influx and actin polymerization. On the other hand, EoL-3 cells show constitutive expression of Fc epsilon RII, Fc gamma RII, LFA-1 and ICAM-1 on their cell surface. The EoL cells may provide new information on some aspects of the signal transduction mechanisms involved in the proliferation, differentiation and activation of eosinophils.

Actins↗

PubMed1993

[Inhibition of eosinophil peroxidase release by chymotrypsin type protease inhibitors from activated human eosinophils].

Eosinophils contain many cytotoxic mediators including eosinophil peroxidase (EPO) in their granules and these mediators are released by various pathophysiological stimuli, resulting in severe damage to various epithelia. However, little is known about the intracellular mechanism of the degranulation. Here we report that eosinophils isolated from patients with bronchial asthma who were not taking corticosteroid hormone had significant amidolytic activities on Suc-Ala-Ala-Pro-Phe-MCA, and also that the activity was completely inhibited by chymostatin (1 x 10(-4) M), eglin C (1 x 10(-4) M), and peptide boronic acid (1 x 10(-4) M), indicating that eosinophils contain a chymotrypsin-like serine protease in the fraction eluted in 50 mM potassium phosphate buffer, pH 8.0 containing 2 M NaCl. Among the various types of protease inhibitors examined, one of the chymotrypsin-type proteases chymostatin (1 x 10(-4) M), but none of the other types of proteases such as leupeptin and E-64, markedly inhibited the EPO release (c.a. 60%) from eosinophils stimulated by immunoglobulin-G (2.5 mg protein/ml beads) in the presence of rhIL-5 (10 ng/ml) or platelet-activating factor (1 x 10(-7) M), although it had no effect on the release by calcium ionophore A23187 (1 x 10(-7) M).

Adult↗

PubMed1993

[The measurement of superoxide generation and eosinophil peroxidase (EPO) from eosinophils].

Eosinophils play an important role in the field of allergy, and release several kinds of granule proteins, leukotrien and superoxide. The most popular method to measure superoxide is utilizing SOD-inhibitable reduction of ferricytochrome c. And recently, the microplate reader enables us to measure superoxide generation from eosinophils more easily using this method. To measure superoxide, we must pay attention to obtain a high purity of eosinophils and not to damage cells. It is also possible to measure eosinophil peroxidase (EPO) using the microplate reader. We are measuring EPO with colorimetric assay using OPD (o-phenylenediamine). OPD is specific to EPO and has no cross reaction to myeloperoxidase. We, now have numerous methods to evaluate eosinophil function, so a selection must be made to select the most correct method.

Cells, Cultured↗

PubMed1995

[Changes in serum eosinophil cation protein level and blood eosinophil density in patients with asthma].

To study the functional state as well as the relation-ship between eosinophil and T lymphocyte in the asthmatics, the authors measured the eosinophil density of blood with percoll, the level of serum eosinophil cation protein (ECP) with radioimmunoassay and soluble IL-2 receptor (sIL-2R) concentration with ELISA. The results revealed that the proportion of hypodense eosinophil and the levels of serum ECP and sIL-2R were 36.3% +/- 14.80%, 10.74 +/- 7.22 mg/L and 0.33% +/- 10 U/L respectively in asthmatics, being significantly higher than those in normal subjects (14.71% +/- 8.20%, 6.93 +/- 4.91 mg/L, 0.13 +/- 0.03 U/L, respectively, P < 0.05 in all). There is remarkable correlation between any two of the three parameters (r = 0.40, 0.37, 0.39, respectively, P < 0.05). It is suggested that eosinophil activation is associated with T lymphocyte activation, both may be involved in the pathogenesis of asthma.

Adult↗

PubMed1997

[The effect of salmeterol on the number of hypodense eosinophils, eosinophilic cationic protein levels ECP in serum, bronchial hyperresponsiveness and lung function test in patients with bronchial asthma].

UNLABELLED: The aim of the study was to compare the influence of salmeterol xinofoate on blood hypodense eosinophil numbers ECP level in serum and sputum, and on lung function tests (VC, FEV1), bronchial hyperresponsiveness (PC20) Lung function tests (FEV1, VC), bronchial histamine challenge (PC20) and measurements of hypodense eosinophils number and ECP level in sputum and serum were done before two and six weeks after treatment with 50 micrograms salmeterol twice daily. Hypodense eosinophils were measured by Kuo et all, ECP by radioimmunosorbent assay using ECP RIA kit Pharmacia. Normal range of ECP is 2.3-15 micrograms/L Hyperreactivity by standard bronchial provocation techniques. The lung function tests (VC, FEV1, FEV1%VC) were measured with Pneumoscreen. RESULT: Eosinophil number decrease after treatment: 42.7 +/- 18.3 before vs 23.4 +/- 8.7 after. ECP level had decreased in serum 53 +/- 24 micrograms/L before and 32 +/- 12 micrograms/L after 2 weeks and 36 +/- 13 micrograms/L after 6 weeks: in sputum 2345 +/- 489 micrograms/g before and 1342 +/- 894 micrograms/g after 2 weeks 1741 micrograms/g after 6 weeks. Lung function tests had improved during the observation and hyperresponsiveness had decreased (0.08 +/- 0.1 microgram/ml before 0.24 +/- 0.2 microgram/ml after 2 week sand 0.4 +/- 0.16 after 6 weeks. CONCLUSION: ECP in serum and sputum in asthmatic patients is higher then in normal patients. Salmeterol improved lung function tests, decreased bronchial responsiveness, decreased hypodense eosinophils number and ECP concentration in serum and sputum. CLINICAL IMPLICATIONS: This date suggested that salmeterol modified bronchial responsiveness and protected the bronchial inflammation.

Adult↗

PubMed1998

Regulation and processing of a precursor form of eosinophil granule major basic protein (ProMBP) in differentiating eosinophils.

The cDNA for eosinophil granule major basic protein (MBP) encodes a prepromolecule with a total length of 222 amino acids (preproMBP). PreproMBP includes a secretory leader of 15 amino acids, an acidic propiece of 90 amino acids, and a basic MBP portion of 117 amino acids. The function of the propiece, which has a predicted pI of 3.9, is unknown, but it gives proMBP an overall acidic charge. Because proMBP is not found in mature eosinophils, we analyzed eosinophil differentiation in interleukin-5 (IL-5)-stimulated umbilical cord stem cells cultured for 24 days. By immunofluorescence, proMBP appeared by day 6 and peaked on day 18, whereas MBP was prominent at days 12 to 24. By day 6, Western blots detected heterogeneous glycosylated 33-kD proMBP; its peak expression occurred on day 12. Western blots showed sequential processing of 33-kD proMBP to an 18-kD intermediate form and finally to 14-kD MBP. By dual label immunoelectron microscopy, proMBP was localized primarily to large uncondensed eosinophil granules, whereas MBP was localized to granules containing a condensed central area. Thus, proMBP is likely expressed and processed as the granule condenses in a multistep process to 14-kD MBP in differentiating eosinophils.

Amino Acid Sequence↗

PubMed1993

[Eosinophil granule proteins (MBP, ECP, EPX/EDN, EPO)--a possible process of eosinophil activation and degranulation].

From current information, a brief review was made on the basic properties of a possible process of eosinophil activation and degranulation. The "activated" eosinophils show the following characteristics: diminished cell density, morphologic alterations, increased surface receptors, heightened parasite killing, increased metabolic activity and prolonged survival. Immune complexes (secretory IgA, IgG, IgE) are known as potent triggering stimuli of eosinophil degranulation as well as complement fragments (C3b, C3bi). Cytokines (IL-5, GM-CSF), PAF and peptides (substance P) act both as weak degranulation inducer and degranulation enhancer. Synergism between the two pathways, Ca2+ and protein kinase C, is now recognized as a common feature of control of secretion in eosinophils.

Animals↗

PubMed1990

Eosinophil-eicosanoid interactions: inhibition of eosinophil chemotaxis in vivo by a LTD4-receptor antagonist.

Chemotaxis of guinea-pig eosinophils in vivo has been studied in the guinea-pig conjunctiva. Guinea-pig eosinophils were labelled with 111In oxine and injected i.v. into recipient animals. Circulating radioactivity remained constant over a period of 4 h but dropped transiently in response to a bolus injection (1 nmol) of platelet activating factor (PAF), suggesting that the 111In-labelled eosinophils had retained their responsiveness to PAF. Leukotriene D4 (LTD4, 1 nmol/eye) and PAF (10 nmol/eye), but not histamine (5 nmol/eye), induced a significant 2.5-fold increase in conjunctival radioactivity, a measure of eosinophil chemotaxis in vivo, after 17 h. Antigen challenge (250 micrograms/eye) in ovalbumin-sensitized animals produced larger responses (8-fold increases) at the same time point. A specific LTD4 receptor antagonist MK-571 (10 micrograms/eye) completely inhibited in vivo chemotactic responses to LTD4, and partially inhibited (54%) the responses to ovalbumin (observations subsequently confirmed by histological studies). As eosinophils may play an important role in allergic diseases, the results with MK-571 indicate that selective and potent LTD4 receptor antagonists may provide a novel therapy for allergic conjunctivitis, rhinitis and asthma.

Animals↗

PubMed1982

The Gordon phenomenon induced by the eosinophil cationic protein and eosinophil protein X.

Cerebellar Purkinje cell degeneration after intracerebral injection of eosinophil granulocytes or extracts thereof is known as the Gordon phenomenon. The reaction is said to be highly selective. An eosinophil-derived neurotoxin (EDN) has recently been reported to induce the Gordon phenomenon. However, we report here that two eosinophil-derived proteins, eosinophil cationic protein (ECP) and eosinophil protein X (EPX), may induce the Gordon phenomenon after intraventricular injection. The potency of ECP is far greater than that of EPX and the latter is possibly identical to EDN. The Fink-Heimer staining for degenerating nerve fibers and boutons, however, indicated that the selectivity of the Gordon phenomenon is not as specific as was previously thought, since this method revealed degeneration of all brain areas in proximity to the ventricular system.

Animals↗

PubMed1987

Ultrastructural and immunohistochemical evidence for release of eosinophilic granules in vivo: cytotoxic potential in chronic eosinophilic pneumonia.

We studied a patient with relapsing idiopathic chronic eosinophilic pneumonia. Each one of four attacks observed during a 4-year period was characterized by dyspnea, wheezing, peripheral blood eosinophilia, elevated serum IgE levels, and pulmonary infiltrates. Glucocorticoid therapy caused prompt resolution of symptoms and disappearance of blood eosinophilia and pulmonary shadowing. Electron microscopy and tissue immunofluorescence studies for major basic protein documented extensive eosinophil degranulation and the presence of free granules within the pulmonary microvasculature. In addition, exfoliation of the alveolar lining cell was observed in association with a clustering of free intact and disintegrating extracellular eosinophil granules against a denuded basement membrane. These findings suggest a cytotoxic potential of the eosinophil at the level of the pulmonary parenchyma in a case of idiopathic chronic eosinophilic pneumonia.

Cytoplasmic Granules↗

PubMed2005

Total eosinophil cationic protein levels in induced sputum as a marker of changes in eosinophilic inflammation in a patient with allergic asthma.

BACKGROUND: Counting eosinophils in induced sputum seems to be a better way to survey disease activity than indirect clinical assessments of airway inflammation, such as pulmonary function measurement. The conventional analysis of induced sputum, however, is time-consuming and requires skilled personnel, which have restricted its use. OBJECTIVE: To determine whether measuring eosinophil cationic protein (ECP) levels in the entire sputum sample could replace the method of counting eosinophils and measuring ECP levels in the supernatant only to establish a clinically more applicable method of studying the intraindividual changes in eosinophilic activity in induced sputum. METHODS: In 13 patients with mild, nonsymptomatic allergic asthma, sputum was induced before and 24 hours after allergen challenge. The entire sputum sample was diluted with 0.1% dithiothreitol, incubated, and divided into 2 parts. One part was processed according to the conventional method, and released ECP levels in the supernatant were measured. The second part was treated with a lysing reagent. Cell debris was separated, and total (intracellular and extracellular) ECP levels in the solution were measured. RESULTS: We found good correlation between total ECP levels in the entire sputum sample and released ECP levels in the supernatant before (r = 0.97) and 24 hours after (r = 0.99) allergen challenge (P < .01 for both). We also found a good correlation between the changes in total and released ECP levels (r = 0.99; P < .01). CONCLUSIONS: Total ECP concentration seems to reflect the eosinophilic inflammatory changes in asthma and might be a useful tool in clinical practice.

Adult↗

PubMed1997

Eosinophilic interleukin 5 (IL-5) transgenic mice: eosinophil activity and impaired clearance of Schistosoma mansoni.

Eosinophilia is a feature common to many invasive helminth infections and eosinophils are often considered to be effector cells in immunity to helminths. This study examined the possible influence of constitutive eosinophilia on the clearance of Schistosoma mansoni infections in mice. Eosinophils from interleukin-5 transgenic mice exhibit normal ultrastructure and function with regard to phagocytosis and killing of bacteria and responses to chemotactic stimuli. IL-5 transgenics and non-transgenic littermates were immunized once or four (hyperimmunization) times with irradiated cercariae of S. mansoni. Animals were challenged percutaneously with unirradiated cercariae one month after their last exposure to irradiated parasites. One month after challenge transgenic animals, whether unimmunized, vaccinated or hypervaccinated, carried significantly more liver-stage parasites than non-transgenic animals. These results suggest that although eosinophils from IL-5 transgenic mice are functional for a number of key parameters, large numbers of eosinophils and/or high levels of IL-5 may in some way impair clearance of S. mansoni. A re-evaluation of the roles of eosinophils and IL-5 in infections with this and other parasites may therefore be warranted.

Animals↗

PubMed1978

High resolution autoradiographic studies of RNA, protein and DNA synthesis during human eosinophil granulocytopoiesis: evidence for the presence of RNA on or within eosinophil granules.

Human bone marrow cells which had been incubated with [3H]uridine or [3H]leucine for I h were studied using the technique of electron microscope-autoradiography. The autoradiographs revealed the presence of newly-synthesized RNA and protein molecules within or on a proportion of (I) the primary and secondary granules in all classes of eosinophil precursors and (2) the secondary granules in eosinophil granulocytes. It is suggested that the granule-associated RNA molecules may be concerned with the synthesis of at least some of the new protein molecules which were incorporated into the limiting membrane or substance of eosinophil granules long after the immature primary granule stage. Studies of eosinophil precursors which had been incubated with [3H]thymidine for I h showed that the eosinophil granules do not label with this DNA precursor.

Autoradiography↗

PubMed2006

Eosinophils in the esophagus--peptic or allergic eosinophilic esophagitis? Case series of three patients with esophageal eosinophilia.

OBJECTIVES: Scattered eosinophils in the distal esophagus traditionally provide the hallmark for peptic esophagitis, but the upper limit of eosinophils and the longitudinal extent of peptic inflammation along the esophagus are unknown. Recently, adults and children with upper intestinal symptoms and >20 eosinophils/high-power field (eos/HPF) have been given the diagnosis of allergic esophagitis. Standardized diagnostic criteria for allergic esophagitis are lacking and the isolated finding of large numbers of eosinophils in the squamous epithelium has been used as the defining feature. We cared for three patients with symptoms and endoscopic features of esophagitis with >20 eos/HPF in their esophageal mucosa. Symptoms, endoscopic features, and histologic findings resolved after 2 months of proton pump inhibitor (PPI) treatment. The aim of this case series is to demonstrate that features thought to be consistent with a diagnosis of allergic esophagitis are also observed in peptic esophagitis. METHODS: A retrospective chart review of three patients with esophagitis (>20 eos/HPF) whose symptoms and eosinophilia resolved with PPI treatment was performed. Esophageal biopsies were reviewed in a blinded manner by one pathologist. RESULTS: Patients (aged 14, 25, and 5 yr) presented with dysphagia, food impaction, and vomiting. Endoscopic features included white exudates and linear furrows. None of the patients received antiallergic treatments or dietary eliminations prior to endoscopy. Following treatment with PPIs alone, all patients became asymptomatic and endoscopic findings reverted to normal. In all three patients, pre- and post-PPI treatment eosinophil numbers/HPF decreased to normal/near normal (37 to 1, 21 to 3, and 52 to 0 eosinophils/HPF in patients 1, 2, and 3, respectively). CONCLUSION: Large numbers of eosinophils can be seen in peptic esophagitis. This histologic finding must be interpreted in the context of the clinical setting in which it is obtained.

Adolescent↗

PubMed2002

Occupational asthma due to low molecular weight agents: eosinophilic and non-eosinophilic variants.

BACKGROUND: Despite having a work related deterioration in peak expiratory flow (PEF), many workers with occupational asthma show a low degree of within day diurnal variability atypical of non-occupational asthma. It was hypothesised that these workers would have a neutrophilic rather than an eosinophilic airway inflammatory response. METHODS: Thirty eight consecutive workers with occupational asthma induced by low molecular weight agents underwent sputum induction and assessment of airway physiology while still exposed at work. RESULTS: Only 14 (36.8%) of the 38 workers had sputum eosinophilia (>2.2%). Both eosinophilic and non-eosinophilic groups had sputum neutrophilia (mean (SD) 59.5 (19.6)% and 55.1 (18.8)%, respectively). The diurnal variation and magnitude of fall in PEF during work periods was not significantly different between workers with and without sputum eosinophilia. Those with eosinophilia had a lower forced expiratory volume in 1 second (FEV1; 61.4% v 83% predicted, mean difference 21.6, 95% confidence interval (CI) 9.2 to 34.1, p=0.001) and greater methacholine reactivity (geometric mean PD20 253 microg v 1401 microg, p=0.007). They also had greater bronchodilator reversibility (397 ml v 161 ml, mean difference 236, 95% CI of difference 84 to 389, p=0.003) which was unrelated to differences in baseline FEV(1). The presence of sputum eosinophilia did not relate to the causative agent, duration of exposure, atopy, or lack of treatment. CONCLUSIONS: Asthma caused by low molecular weight agents can be separated into eosinophilic and non-eosinophilic pathophysiological variants with the latter predominating. Both groups had evidence of sputum neutrophilia. Sputum eosinophilia was associated with more severe disease and greater bronchodilator reversibility but no difference in PEF response to work exposure.

Asthma↗