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PubMed1981

Tetrapeptides of the eosinophil chemotactic factor of anaphylaxis (ECF-A) enhance eosinophil Fc receptor.

Certain mast cell products, including the tetrapeptides Ala-Gly-Ser-Glu (Ala4) and Val-Gly-Ser-Glu (Val4) of the eosinophil chemotactic factor of anaphylaxis (ECF-A) and histamine, are preferentially chemotactic for eosinophils in vitro. Amino acid substitutions or modifications of the ECF-A tetrapeptides may substantially alter the chemotactic activity of human eosinophils. The cellular mechanism whereby ECF-A tetrapeptides enhance IgG-dependent killing of schistosomula by rat eosinophils is, however, unclear. We now report that the parent tetrapeptides and the substituted analogue Val-Pro-Ser-Glu (Pro3), but not histamine, increase the number of rat and human eosinophils that form rosettes with erythrocytes bearing IgG antibodies. Moreover, we find a correlation between the effect of these substances on the expression of rat eosinophil IgG Fc receptors and their capacity to increase the IgG-dependent cytotoxicity of rat eosinophils for Schistosoma mansoni schistosomula.

Animals↗
PubMed2002

Accuracy of eosinophils and eosinophil cationic protein to predict steroid improvement in asthma.

BACKGROUND: There is a large variability in clinical response to corticosteroid treatment in patients with asthma. Several markers of inflammation like eosinophils and eosinophil cationic protein (ECP), as well as exhaled nitric oxide (NO), are good candidates to predict clinical response. AIM: We wanted to determine whether we could actually predict a favourable response to inhaled corticosteroids in individual patients. METHODS: One hundred and twenty patients with unstable asthma were treated with either prednisolone 30 mg/day, fluticasone propionate 1000 microg/day b.i.d. or fluticasone propionate 250 microg/day b.i.d., both via Diskhaler. They were treated during 2 weeks, in a double-blind, parallel group, double dummy design. We measured eosinophils and ECP in blood and sputum, and exhaled nitric oxide as inflammatory parameters before and after 2 weeks in order to predict the changes in forced expiratory volume in 1 s (FEV1), provocative concentration of methacholine causing a 20% fall in FEV1 (PC20 Mch), and asthma quality of life (QOL). Secondly, to test whether these results were applicable in clinical practice we determined the individual prediction of corticosteroid response. RESULTS: We found that changes in FEV1, PC20 Mch and QOL with corticosteroids were predominantly predicted by their respective baseline value and to a smaller extent by eosinophils in blood or sputum. ECP, measured in blood or sputum, was certainly not better than eosinophils in predicting clinical response to corticosteroids. Smoking status was an additional predictor for change in FEV1, but not for change in PC20 Mch or QOL. Prediction of a good clinical response was poor. For instance, high sputum eosinophils (> or = 3%) correctly predicted an improvement in PC20 Mch in only 65% of the patients. CONCLUSION: Our findings show that baseline values of the clinical parameters used as outcome parameters are the major predictors of clinical response to corticosteroids. Eosinophil percentage in blood or sputum adds to this, whereas ECP provides no additional information. Correct prediction of clinical response in an individual patient, however, remains poor with our currently used clinical and inflammatory parameters.

Adolescent↗
PubMed1997

Production of IL-8 and release of eosinophil-derived neurotoxin by normal peripheral blood eosinophils.

To study the regulation of IL-8 production and release of eosinophil-derived neurotoxin (EDN) by normal blood eosinophils, we isolated eosinophils from healthy individuals and stimulated them with immobilized secretory IgA with or without exogenous IL-5 for 3, 12, and 24 h, and with different concentrations of ionomycin for 24 h. Eosinophils cultured with secretory IgA for 3 and 12 h showed strong expression of mRNA for IL-8 by reverse transcription polymerase chain reaction. Exogenous IL-5 enhanced mRNA for IL-8 expression by eosinophils after 24 h of incubation. IL-8 secretion increased in a time-dependent manner throughout the 24 h of observation; in contrast, EDN release reached a plateau value after 12 h. Furthermore, at least 2 microM ionomycin was necessary for induction of IL-8 secretion, whereas 0.5 microM induced detectable EDN release by eosinophils. These results suggest that the mechanism of eosinophil degranulation may be different from those of cytokine secretion.

Dose-Response Relationship, Drug↗
PubMed1994

Eosinophil and eosinophil cationic protein in allergic rhinitis.

To evaluate the roles of eosinophils in allergic rhinitis, eosinophil cationic protein (ECP) levels in sera and nasal secretions were measured in 28 patients with allergic rhinitis. In patients, blood eosinophil counts and serum ECP levels were significantly higher than in controls (p < 0.05). However, there was no statistically significant correlation between blood eosinophil count and serum ECP level (r = 0.025). The number of eosinophils and the ECP level in nasal secretion were also measured. In patients, the ECP level increased with the number of eosinophils, and the ECP level in nasal secretion was significantly higher than in controls (p < 0.05). These results suggest that eosinophils and ECP play important roles in allergic rhinitis.

Adolescent↗
PubMed1994

Effect of clotting temperature and eosinophil concentration on the eosinophil cationic protein concentration in serum.

Earlier investigations have indicated the need for detailed instructions about the incubation conditions of blood samples before establishment of eosinophil cationic protein concentration in serum. Therefore, the effect of different clotting temperatures and eosinophil concentrations on the serum eosinophil cationic protein concentration was quantified in 40 subjects. Our results show that serum eosinophil cationic protein concentrations strongly depend on the clotting temperature. Blood samples clotted for 1 h at 37 degrees C had 5-10 times higher serum eosinophil cationic protein concentrations than blood samples of the same subject clotted for 1 h at 0 degrees C. Higher eosinophil counts resulted also in increased serum eosinophil cationic protein concentrations.

Blood Coagulation↗
PubMed2001

The serum levels of eosinophil cationic protein (ECP) are related to the infiltration of eosinophils in the tumours of patients with Hodgkin's disease.

We have previously described a relation between abundance of eosinophilic granulocytes in Hodgkin's disease (HD) tumours and poor prognosis. In order to further explore the importance of the eosinophilic infiltration, we immunohistochemically examined the presence of eosinophils, using the monoclonal antibodies EG 1 and EG 2, in the tumours of 54 newly diagnosed patients with HD and related the degree of infiltration to clinical characteristics and the serum levels of eosinophil cationic protein (S-ECP). S-ECP levels (upper normal value 16 micrograms/l) varied between 2.2 and 71.7 micrograms/l, mean 25.4 micrograms/l. There was an association (p = 0.01) between the number of eosinophils in the tumour tissue and S-ECP. S-ECP levels were also associated to high erythrocyte sedimentation rate (ESR, p < 0.01) and nodular sclerosis (NS) histology (p < 0.05), and there was a tendency of a correlation to bulky disease (p = 0.06). The number of eosinophils stained with EG 2 correlated to high ESR (p < 0.05), and to high leukocyte count (p = 0.02). A follow-up value of S-ECP after treatment was, in most of the cases measured, lower than the initial value. The high values of S-ECP in several patients with HD probably originates from eosinophils infiltrating the tumours. The same patients had a higher ESR and tended to have a more advanced stage and bulky disease. There are no significant correlations with disease-free and overall survival, as the follow-up time is short, and prognosis favourable.

Adolescent↗
PubMed2003

Nitric oxide metabolites, eosinophils, and eosinophilic cationic protein in patients with asthma: sputum versus blood.

The monitoring of airway inflammation has assessed in bronchial asthma directly by sputum examination, and indirectly by measurements in peripheral blood. To investigate the diagnostic value of these two methods, we compared nitric oxide (NO) metabolites, eosinophils, and eosinophil cationic protein (ECP) in sputum and blood in patients with asthma and control subjects. Sputum and serum were obtained from fifteen patients with asthma, and then were examined before anti-asthma treatment, including steroid preparations. ECP was measured by fluoroimmunoassay. NO metabolites were assayed by using modified Griess reaction. Asthmatic patients, compared with control subjects, had significantly higher level of NO metabolites, higher proportion of eosinophils, and higher levels of ECP in sputum. Asthmatic patients, compared with control subjects, however, had significantly higher number of eosinophils, and were at higher levels of ECP in blood. FEV1, FEV1/FVC was negatively correlated with sputum eosinophils. The area under receiver operating characteristic (ROC) curve showed that eosinophils in sputum are significantly accurate markers than NO metabolites in sputum and blood. These findings suggest that the proportion of eosinophils in sputum have more accurate diagnostic marker of asthmatic airway inflammation than NO metabolites in sputum in differentiating asthmatic patients from control subjects.

Adult↗
PubMed1998

[Eosinophilic activation based on measurements of eosinophil cationic protein in patients with acute myocardial infarction and patients with angina pectoris].

Increased activation of polymorphonuclear neutrophils in the wideness of irreversible myocardial injury was described by many authors. Released proteolytic enzymes may cause deliquescence of necrotic muscle tissue and attenuate collagenic structure of myocardium, lead to endothelium damage and generate free oxygen radicals. Eosinophilic leucocytes reveal also enhanced proinflammatory activation. They take participation in inflammatory and immunologic reactions, among others, by producing and releasing many biologic mediators. One of the most important and powerful mediators is eosinophil cationic protein (ECP), the specific marker of eosinophil activation in vivo. We studied 17 patients (pts) with acute myocardial infarction (MI) and 21 pts with angina pectoris (AP). The plasma concentrations of ECP and the number of eosinophils in peripheral blood were measured 3 times-near before beginning of the treatment, and on the 4-th and 8-th day of MI. During the first 4 days measurements of CK-MB every 6 hours were made. We observed the significant increase of eosinophils and ECP correspondingly on the 8-th and 4-th day, when compared to the first day of MI (p < 0.05) and the patients with AP (p < 0.01). Despite tendency, the significant correlation of eosinophils and ECP values was not obtained (r = 0.36). In the group of patients with AP the eosinophil and ECP values were significantly higher at the pts with unstable angina, when compared to pts with stable angina (p < 0.001).

Angina Pectoris↗
PubMed1986

Monoclonal antibodies to human eosinophil plasma membrane antigens enhance the secretion of eosinophil cationic protein.

This study was done to examine the nature of the membrane constituents involved in the secretion of eosinophil cationic protein (ECP) from human blood eosinophils. Three mouse monoclonal antibodies were used, which showed greater binding to membrane antigens on activated, and light density eosinophils from patients with an eosinophilia, than on nonactivated or normal density eosinophils. All three antibodies (EoN4, EoN5 & EoN6) stimulated normal density human eosinophils to secrete ECP, either alone or in association with sepharose-C3b. The antibodies bound to at least two separate sites on the membrane, which were distinct from the receptors for immunoglobulins, C3b, and eosinophil activating factor. One combination of antibodies increased the amount of ECP which was secreted. The membrane antigen recognized by antibody EoN4 was a glycoprotein, molecular weight 75 kD. These findings showed that ECP secretion may be induced by a wider range of stimuli than has been previously recognized, and that the antigens recognized by these monoclonal antibodies may play an important role in the induction of eosinophil degranulation.

Antibodies, Monoclonal↗
PubMed1985

Eosinophil involvement in rheumatoid arthritis as reflected by elevated serum levels of eosinophil cationic protein.

Circulating levels of eosinophil cationic protein (ECP), an eosinophil specific granule protein, and numbers of peripheral eosinophils were determined in 42 patients with rheumatoid arthritis. At the time of the investigation the patients were without drug treatment. They had normal blood counts of eosinophils but on average a five-fold increase of the serum ECP values compared with healthy subjects. The intracellular content of ECP in eosinophils isolated from 14 patients was normal. High serum levels of ECP were particularly observed in patients with a disease of rather short duration but with a more aggressive course. Other factors associated with high ECP values were blood eosinophil counts in the upper normal range, high rheumatoid factor titre and increased inflammatory activity as defined by elevated serum haptoglobin and blood platelet counts. No relation was found between serum ECP and circulating immune complexes or serum total IgE. Synovial fluids obtained from 14 patients with rheumatoid arthritis contained very high concentration of ECP; on average nine times higher than those in the circulation of the patients. During corticosteroid but not NSAID therapy serum ECP decreased on average about 50% compared with pre-treatment values. Although eosinophils are not a notable feature of the synovial membrane infiltrate or cellular joint exudate, data obtained indirectly indicates their participation in the inflammatory reaction in RA.

Anti-Inflammatory Agents↗
PubMed1994

Eosinophil peroxidase-dependent hydroxyl radical generation by human eosinophils.

Eosinophil production of superoxide (O2-.) and hydrogen peroxide (H2O2) is important in host defense. The present study assessed the potential of eosinophils to generate another potent cytotoxic species, the hydroxyl radical (.OH). .OH formation by phorbol myristate acetate (PMA)-stimulated eosinophils was demonstrated using an alpha-(4-pyridyl-1-oxide)-N-tert-butyl nitrone/ethanol spin trapping system. Additionally, .OH was spin trapped following the addition of purified eosinophil peroxidase (EPO) to a cell-free O2-./H2O2 generating systems. Effects of superoxide dismutase, catalase, azide, aminotriazole, chloride-depleted buffer, and extensive metal chelation were consistent with .OH formation via the reaction of O2-. and EPO-generated hypohalous acid. Under chloride-depleted conditions, physiologic concentrations of Br- increased .OH formation by both PMA-stimulated eosinophils and the cell-free EPO system. Physiologic concentrations of SCN-, however, did not increase .OH formation, and in the presence of both Br- and SCN-, .OH formation was similar to SCN- only. Eosinophils appear to form .OH via an EPO-dependent mechanism, the magnitude of which varies with the availability of various EPO substrates. Given the highly reactive nature of this radical and the ability of EPO to adhere to cell membranes, even small amounts of .OH formed at such sites could contribute to eosinophil-mediated cytotoxicity.

Bromides↗
PubMed1997

The eosinophil granule proteins in serum, but not the oxidative metabolism of the blood eosinophils, are increased in cancer.

The eosinophil activity in patients with renal cell adenocarcinoma during treatment with interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) was reduced when measured as zymosan-induced lucigenin-enhanced chemiluminescence (CL). Eosinophil cationic protein (ECP), eosinophil peroxidase (EPO) and eosinophil protein-X (EPX) were significantly elevated before and during treatment (P<0.001) when compared with the controls. ECP and EPO were unaffected by the treatment whereas it induced an increased EPX level compared with values measured before treatment (P<0.05). The propensity of eosinophils to secrete their granule proteins may reflect the fact that eosinophils in cancer patients have an enhanced capacity to kill cancer cells.

Adult↗
PubMed1996

Different effects of cyclic AMP and butyrate on eosinophilic differentiation, apoptosis and bcl-2 expression of a human eosinophilic leukemia cell line, EoL-1.

A human eosinophilic leukemia cell line, EoL-1, stopped proliferating at the G1 phase, differentiated into eosinophilic granule-containing cells, and died by apoptosis when stimulated with dibutyryl cyclic AMP (dbcAMP). To clarify the effects of dbcAMP, the effects of butyrate and cAMP-increasing reagents, prostaglandin E2 (PGE2) and forskolin, on EoL-1 cellular differentiation and apoptosis were examined and compared. PGE2 and forskolin but not butyrate induced differentiation to eosinophilic granule-containing cells, suggesting that cAMP played a primary role in eosinophilic differentiation of EoL-1 cells. PGE2, forskolin and butyrate, when used alone, did not induce apoptosis of EoL-1 cells significantly at the concentrations used, but sequential stimulation of EoL-1 cells with the cAMP-increasing reagents and butyrate showed that butyrate induced further maturation and apoptosis of cAMP-induced eosinophilic granule-containing cells. These results showed that cAMP and butyrate have different effects on eosinophilic differentiation and apoptosis of EoL-1 cells. The cAMP-increasing reagents and butyrate also showed different effects on expression of members of the bcl-2 family; PGE2 decreased bcl-2 and bax levels, whereas butyrate increased the bcl-2 level. PGE2 or PGE2+butyrate, but not butyrate alone, induced bcl-XS expression. EoL-1 cells constitutively expressed Fas and anti-Fas antibody induced EoL-1 cell death, but the Fas/Fas ligand system was not involved in dbcAMP-induced EoL-1 cell apoptosis. The EoL-1 cell line is thus a useful model in which to examine differentiation and apoptosis of eosinophilic leukemia cells.

Apoptosis↗
PubMed2006

[Eosinophil granule cationic proteins: eosinophil activation markers].

INTRODUCTION: Eosinophils play a central role in the pathogenesis of allergic diseases and host protection against parasites, especially against helminths. EXEGESIS: Eosinophil granules contain cytotoxic proteins, including cationic proteins: major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil peroxidase (EPO) and eosinophil derived neurotoxin (EDN). CONCLUSION: Eosinophil granule proteins are implicated in the occurrence of allergic diseases and Churg-Strauss syndrome. These proteins are activation markers of eosinophils and may be useful in clinical practice.

Biomarkers↗
PubMed1998

Bronchoalveolar lavage cell findings in three types of eosinophilic pneumonia: acute, chronic and drug-induced eosinophilic pneumonia.

There are clinically different types of eosinophilic pneumonia (EP) but no study to date has compared pulmonary inflammatory cells between different types of EP, such as acute eosinophilic pneumonia (AEP), chronic eosinophilic pneumonia (CEP) and drug-induced eosinophilic pneumonia (drug-EP). The present study compared bronchoalveolar lavage fluid (BALF) cell findings to elucidate whether the profiles of the pulmonary inflammatory cells were different among the three types of EP. Clinical records of 28 patients with EP, consisting of eight AEP patients, 10 CEP patients and 10 drug-EP patients, were examined retrospectively. The differential cell counts, the CD4+/CD8+ ratio of lymphocytes, the percentage of HLA-DR+ in CD4+ and CD8+ lymphocytes, and the mean number of nuclear segmentations in cosinophils in BALF were compared among the three types of EP. The numbers of total cells, lymphocytes, neutrophils and eosinophils in BALF from patients with AEP were increased compared with those from normal subjects, and patients with CEP and drug-EP. The CD4+/CD8+ ratio of the BALF lymphocytes in patients with AEP, which exceeded 1.0 in all patients, was significantly higher than that in normal subjects. The percentages of HLA-DR+ cells in CD8+ lymphocytes in BALF from patients with CEP were significantly higher than those from patients with AEP and drug-EP. There was no significant difference in the mean number of nuclear segmentations in eosinophils in BALF among the three types of EP. The BALF cell findings in patients with EP showed some characteristics in accordance with type of EP. It is suggested that pulmonary neutrophils and lymphocytes, rather than eosinophils, may be related to the pathogenesis of the different types of EP.

Acute Disease↗
PubMed1999

Overexpression of CD44 on alveolar eosinophils with high concentrations of soluble CD44 in bronchoalveolar lavage fluid in patients with eosinophilic pneumonia.

BACKGROUND: High levels of interleukin-5 (IL-5) are found in the alveolar space of patients with eosinophilic pneumonia (EP). IL-5 promotes the growth and differentiation of eosinophils, as well as activating these cells. IL-5 also induces the expression of CD44 on eosinophils in vitro. To evaluate the contribution of CD44 to the pathogenesis of EP, we examined the expression of CD44 on eosinophils in bronchoalveolar lavage fluid (BALF) and measured the concentration of soluble CD44 (sCD44) in BALF from patients with EP. METHODS: The concentrations of IL-5, sCD44, and hyaluronic acid (HA) were measured in BALF. The expression levels of CD44 on eosinophils in BALF and peripheral blood in patients with EP were compared. RESULTS: The expression of CD44 on alveolar eosinophils and the concentration of sCD44 were increased in BALF of patients with EP. There was a significant correlation between IL-5 and sCD44 in BALF. A high concentration of HA was observed in BALF of EP patients. CONCLUSIONS: Our results suggest that the high expression of CD44 on eosinophils probably results from upregulation by IL-5 and could be important in the pathogenesis of EP.

Adult↗
PubMed2002

Immunohistochemical artifact for nitrotyrosine in eosinophils or eosinophil containing tissue.

Immunohistochemical artifacts for nitrotyrosine were investigated in eosinophils with regard to fixatives. Immunoreactivity for nitrotyrosine was revealed in separated eosinophils and in gastric mucosa fixed with periodate, lysine-paraformaldehyde (PLP). The increase in immunoreactivity by PLP was due to periodate itself, a component of PLP. Nitrotyrosine formed by peroxidase using NO2- and H2O2 or by peroxynitrite was not completely inhibited by 100 mM dithionite but the immunoreactivity for nitrotyrosine antibodies by PLP was completely inhibited by 5.7 mM dithionite. Although untreated eosinophils or ovalbumin (OVA) did not show protein tyrosine nitration in a standard Western blot, the treatment of the blotted membrane with PLP increased the reactivities of proteins from eosinophils with anti-nitrotyrosine antibodies. The increase in immunoreactivity of OVA with anti-nitrotyrosine antibodies by PLP did not change with pre-treatment with dithionite but was abolished by treatment with dithionite after PLP fixation. In HPLC assays, periodate did not generate nitrotyrosine from L-tyrosine and aminotyrosine. These results suggest that the treatment of eosinophils or eosinophil-containing tissues with PLP fixative augments the immunoreactivity of nitrotyrosine antibodies with eosinophils due to the formation of epitopes similar to nitrotyrosine by an oxidation reaction of periodate, which evokes an artifact in nitrotyrosine immunohistochemistry.

Artifacts↗
PubMed1988

Highly purified murine interleukin 5 (IL-5) stimulates eosinophil function and prolongs in vitro survival. IL-5 as an eosinophil chemotactic factor.

The recent molecular cloning of the complementary DNA encoding T cell--replacing factor (TRF) has demonstrated that a single molecule is responsible for B cell growth factor II (BCGF-II) activity and eosinophil differentiation activity. It has been proposed that this molecule be called interleukin 5 (IL-5). We previously reported that purified rIL-5 supports the terminal differentiation and proliferation of eosinophilic precursors. In this study, we examined the effects of IL-5 on functional activities of mature eosinophils. IL-5 maintained the viability of mature eosinophils obtained from peritoneal exudate cells of mice infected with parasites. It also induced superoxide anion production in a dose-dependent manner. The Boyden's chamber Millipore assay revealed that IL-5 had a marked chemokinetic effect on eosinophils in a dose-dependent manner. Moreover, IL-5 was found to be an eosinophil chemotactic factor by the checkerboard assay. In conclusion, IL-5 is suggested to play an important role in increasing the functional activities of eosinophils as well as their production in allergic and parasitic diseases.

Animals↗