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[Serum eosinophil cationic protein, urinary eosinophil protein X and urinary N-methylhistamine during acute asthma in children].

Serum eosinophil cationic protein (ECP), urinary eosinophil protein X (EPX) and urinary N-methylhistamine were investigated in 24 asthmatic children during acute asthma to determine the sequential variation in the parameters. Serum ECP levels and urinary EPX levels were high during acute asthma, and decreased gradually with improvement of acute asthma, however, these levels of eosinophil drived proteins were significantly higher than those in non-atopic children. There was no significant variation of urinary NMH levels. Significant inverse correlation was found between lung function and urinary EPX level. The present study suggests that serum ECP and urinary EPX may be useful for monitoring airway inflammation during acute asthma, and activation of eosinophils may persist after improvement of acute asthma.

Acute Disease↗

Human eosinophil-derived neurotoxin and eosinophil protein X are likely the same protein.

The human eosinophil granule contains a series of cationic proteins. Two of these, eosinophil-derived neurotoxin (EDN) and eosinophil protein X (EPX), are reported to have similar m.w. and both possess neurotoxic and helminthotoxic activities. Therefore, the properties of these molecules were analyzed to determine whether they differ. EDN was purified from eosinophils of patients with the hypereosinophilic syndrome and EPX from the buffy coat cells of normal individuals. By SDS-PAGE, both proteins showed a major band at 18.7 kDa and a minor band at 21.4 kDa. By two-dimensional non-equilibrium gel electrophoresis the proteins migrated identically. With radiolabeled proteins in reverse phase HPLC, both proteins eluted at the same concentration of acetonitrile and showed identical tryptic maps. Both proteins possessed comparable ribonuclease activity and both were comparably neurotoxic in the rabbit. By immunodiffusion the two proteins showed a reaction of identity; by RIA, with both polyclonal and monoclonal antibodies, the proteins had very similar inhibitory activities. These results indicate that EDN and EPX have virtually identical properties and are probably the same protein.

Animals↗

Eosinophilic vasculitis syndrome: recurrent cutaneous eosinophilic necrotizing vasculitis.

We recently identified a syndrome of recurrent cutaneous eosinophilic vasculitis in three patients. These patients had in common widespread pruritic, erythematous, purpuric papules and angioedema of face and hands associated with peripheral blood eosinophilia. Eight skin biopsies from these three patients all showed necrotizing vasculitis of the small vessels of the skin, with exclusively eosinophilic infiltration and minimal or no leukocytoclasis. The disease followed a chronic course, with recurrent, itchy, swelling skin lesions and without evidence of systemic involvement over observation periods of 3, 17, and 23 years. The skin lesions responded promptly to systemic steroid treatment, but two patients required maintenance doses for control of the disease. Immunofluorescence studies showed marked deposition of the cytotoxic eosinophil granule major basic protein in the affected vessel walls. Eosinophil-active cytokine IL-5 was detected in the serum of one patient. Expression of the vascular cell adhesion molecule-1 for eosinophil adherence was detected on the endothelium of the affected vessels. Because this disease showed distinctive clinical manifestations and characteristic histopathological features, we believe it is a distinct entity and should be distinguished from other types of vasculitis.

Adolescent↗

Bullous eosinophilic cellulitis succession with eosinophilic pustular folliculitis without eosinophilia.

Eosinophilic cellulitis is characterized clinically by an acute dermatitis resembling cellulitis with unknown etiology. Eosinophilic pustular folliculitis is also a rare inflammatory dermatosis characterized by recurrent crops of erythematous follicular papulopustules that coalesce to form annular plaques with unclear etiopathogenesis. We describe a 20-year-old white male who had vesiculobullous and plaque-like lesions on the hands and feet and was diagnosed with bullous eosinophilic cellulitis clinically and histologically without any etiological agents. Following therapy with oral corticosteroid and oral tetracycline capsules, the lesions disappeared. After a 2-month asymptomatic period, the patient developed pruritic follicular papules and pustules on the lower and upper extremities and upper back. Stool examination revealed Gierdia intestinalis eggs. The patient had complete clearance with treatment of ornidazol for 2 weeks and indomethacin for 2 months. This is the first report of bullous eosinophilic cellulitis coexisting with eosinophilic pustular folliculitis without eosinophilia in the English published work.

Adult↗

Transient wheeze. Eosinophilic bronchobronchiolitis in acute eosinophilic pneumonia.

The clinicopathologic features of five patients with acute eosinophilic pneumonia who presented with transient wheeze as well as acute onset of high fever, severe hypoxemia, and diffuse pulmonary infiltrates are described. Eosinophilic pneumonia was diagnosed by bronchoalveolar lavage and transbronchial lung biopsy. The illness resolved rapidly with or without corticosteroid therapy. No relapse occurred. To characterize the transient wheeze, a transbronchoscopic bronchial biopsy and pulmonary function tests were performed. Specimens of bronchial wall revealed eosinophil infiltration into the bronchial mucosa. Pulmonary function tests demonstrated reduced diffusing capacity and small airway dysfunction. These findings suggested that eosinophil infiltration into the bronchial mucosa might temporarily cause transient wheeze, different from bronchial asthma, due to small airway dysfunction in acute eosinophilic pneumonia.

Acute Disease↗

[Eosinophilic gastroenteritis: an example of gastrointestinal toxicity of eosinophils].

Toxicity of polynuclear eosinophils in the digestive tract results from a cascade of immune responses involving various mediators including mastocytes and T-lymphocyte helpers. Polynuclear eosinophils may be implicated in many digestive disorders. This involvement is well established in eosinophilic gastroenteritis which has become the model for study of eosinophil toxicity on the digestive tract. Considered to be of uncertain etiology up until 1998, this mechanism is well illustrated by the case described in this report involving infiltration of all layers of the small intestinal mucosa in a 23-year-old patients who responded well to corticosteroid treatment. This case provides the opportunity to update physiopathologic and therapeutic data concerning this unpredictable syndrome. Current evidence of transition forms from idiopathic hypereosinophilic syndrome casts doubt on the conventional classification of eosinophilic gastroenteritis and underlines the need for careful diagnostic work-up and prolonged follow-up.

Abdominal Pain↗

Atopic aspect of eosinophilic nasal polyposis and a possible mechanism of eosinophil accumulation.

Eosinophil-infiltrated nasal polyps (eosinophilic polyps) have been increasing in Japan. This increase is parallel with an increase in cases of nasal allergy, and is an interesting contrast to the decrease in chronic infectious nasal diseases. Thus IgE-mediated hypersensitivity could be considered an etiologic factor of eosinophilic polyposis. However only 29.7% of patients with eosinophilic polyposis were defined as being atopic. In this study, high levels of IgG-immune complex were found in eosinophilic polyp fluid. It is conceivable that the IgG-immune complex may play some part in tissue eosinophilia of polyps.

Antigen-Antibody Complex↗

Serum levels of eosinophil cationic protein and eosinophils in asthmatic children during a course of prednisolone therapy.

Eosinophils play an important role in the inflammatory events of allergic asthma. Serum eosinophil cationic protein (ECP) is a marker of disease activity and of treatment efficacy in bronchial asthma. To understand the role of ECP concentrations in disease activity of acute asthma, we determined changes in serum concentrations of ECP elaborated by activated eosinophil before and after prednisolone therapy. Circulating levels of ECP in 15 normal control subjects, and in sera of 20 asthmatic children who were allergic to house dust mites, were measured during an acute exacerbation and when the children were in stable condition, using commercially available assay kits. The mean concentrations of serum ECP were significantly higher during an acute asthma exacerbation than when the children were stable (26.41 +/- 21.66 microg/L vs 15.74 +/- 11.36 microg/L P < 0.01) or when compared to control subjects (7.50 +/- 1.42 microg/L; P < 0.001). The mean eosinophil counts (EC) during acute asthma attacks (575 +/- 286/mm3) and when stable (467 +/- 204/mm3) were higher than in the control group (181 +/- 164/mm3). The differences were statistically significant among the three groups (P < 0.05). A significant correlation was found between serum levels of ECP and EC (r = 0.788, P = 0.001) in asthmatic children; there were also significant correlations between ECP and EC in nonallergic normal control subjects (r = 0.662; P = 0.007). In conclusion, this study provides further evidence that changes in serum ECP may serve as an objective indicator for clinical activity and results of treatment in allergic asthmatics.

Adolescent↗

C/EBP regulates the promoter of the eosinophil-derived neurotoxin/RNS2 gene in human eosinophilic cells.

The eosinophil-derived neurotoxin (EDN), a member of the mammalian ribonuclease family, is found in the large specific granules of human eosinophilic leukocytes. We have investigated the role of the C/EBP transcription factor family in the regulation of EDN promoter activity. Here we show that the C/EBP family is involved in intrinsic regulation of EDN promoter activity. We have identified a C/EBP binding site located at -124 in the proximal promoter of the EDN gene. Mutation of this C/EBP site results in a decrease of promoter activity in HL-60-eos cells as well as in eosinophils differentiated in vitro from CD34+ cells. Different C/EBP proteins are able to bind to the C/EBP site as shown by gel shift assay. Our results indicate the importance of the C/EBP family in the regulation of the EDN gene in eosinophils.

Animals↗

In vitro release of eosinophil cationic protein from peripheral eosinophils reflects disease activity in childhood Crohn disease and ulcerative colitis.

UNLABELLED: The aim of this study was to compare in vitro release of eosinophil cationic protein (ECP) from peripheral blood eosinophils during active phases of childhood Crohn disease (CD) and ulcerative colitis with phases of remission. Ten children with CD and nine children with ulcerative colitis were investigated during 55 and 56 clinical visits, respectively. Each patient was investigated during at least one phase of clinically active disease and one phase of remission. Disease activity was assessed by means of the Paediatric Crohn Disease Activity Index (PCDAI) in Crohn disease and according to the clinical activity index of Rachmilewitz in ulcerative colitis. On an intra-individual basis, in vitro ECP release was significantly higher (P < 0.001) during active phases of CD and ulcerative colitis than in phases of remission (CD:median: 24.5 microg/l, range 16.0-61.2 versus median: 5.7 microg/l, range 2.0-16.7; ulcerative colitis: 14.8 microg/l, 8.3-39.8 versus 4.9 microg/l, 2.0-9.9). On an inte-individual basis, in CD and ulcerative colitis a strong and highly significant correlation was observed between disease activity indices and in vitro release of ECP from peripheral eosinophils (r = 0.89, P < 0.0001 and r = 0.82, P < 0.0001, respectively). CONCLUSION: These results show that in vitro ECP release from peripheral eosinophils reflects disease activity in both CD and ulcerative colitis and therefore can be used as a new appropriate laboratory parameter for assessment of disease activity in chronic inflammatory bowel disease.

Adolescent↗

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↗

Measurement of eosinophil cationic protein (ECP) and eosinophil protein X/eosinophil derived neurotoxin (EPX/EDN). Time and temperature dependent spontaneous release in vitro demands standardized sample processing.

The measurement of eosinophil derived proteins such as eosinophil cationic protein (ECP) and eosinophil protein X/eosinophil derived neurotoxin (EPX/EDN) in biological fluids may be a useful indicator of eosinophil activity in ongoing inflammatory processes. This study was performed on blood samples and illustrates that serum values of ECP in particular, but also of EPX, are mainly a result of spontaneous release during the processing of blood samples. In the presence of divalent cations (Ca2+ and Mg2+), the amount of released ECP and EPX is dependent upon the incubation temperature and the time before centrifugation and recovery of serum. Moreover, the utensils used for blood sampling may influence the serum levels of ECP and EPX. Thus, standardized sample processing is of paramount importance if the results are to have optimal diagnostic or clinical value.

Blood Proteins↗

Activated eosinophils in familial eosinophilic gastroenteritis.

Two siblings with eosinophilic gastroenteritis who presented with severe iron deficiency anemia and hypoalbuminemia associated with varying degrees of mucosal damage are described. Using a monoclonal antibody to the secreted form of eosinophil cationic protein, we demonstrated activated degranulating eosinophils in the gastrointestinal mucosa that correlated with the degree of histologic damage. This finding suggests that eosinophils may have a primary role in the tissue damage in this condition. Oral disodium cromoglycate (Nalcrom) failed to alter the clinical, radiologic, and histologic abnormalities. Prednisolone, however, was beneficial in 1 case but did not completely reverse the abnormalities.

Adolescent↗

Increase of mast cells may be associated with infiltration of eosinophils and proliferation of microvessels in gastric eosinophilic granuloma.

BACKGROUND: Gastric eosinophilic granuloma (GEG) is a rare disease. Recently this disease has begun to increase in China. In the present study, the function and the role of mast cells (MC) in the pathogenesis of GEG were investigated. METHODS: Paraffin-embedded tissue sections from 23 GEG patients and 15 gastric ulcer (GU) patients, were stained with antihuman mast cell tryptase for counting the MC and degranulated MC. Antihuman CD34 antibody was used for detecting the microvessel density (MVD) with immunohistochemical technique. Mast cell degranulation was also studied using electron microscopy. RESULTS: The quantity of both MC and degranulated MC were higher in both GEG and GU than in normal gastric mucosa. The proportion of degranulated MC was higher in the GEG but in GU it was similar to normal mucosa. The MVD was higher in both GU and GEG than that in the normal gastric mucosa and it was higher in the high-MC group than in the low-MC group in GEG. The positive correlation between eosinophil and MC was present only in GEG, not in GU. CONCLUSIONS: The infiltration of eosinophils and MVD may be associated with the increase of MC in GEG. This suggests that in addition to eosinophils, MC might be the important cells in the pathogenesis of GEG.

Adult↗

Activation of eosinophils and fibroblasts assessed by eosinophil cationic protein and hyaluronan in BAL. Association with acute rejection in lung transplant recipients.

Lung transplantation has become an accepted therapy for end-stage lung disease. Acute rejection of the transplanted hung still remains a major clinical problem since it decreases graft survival. Eosinophil cationic protein (ECP) from activated eosinophils, hyaluronan (HYA) from fibroblasts, and circulating intercellular adhesion molecule 1 (1CAM-1) have been associated with acute rejection in kidney and liver grafts. We investigated whether these, as well as other molecules, were increased in acute rejection of lung allografts. Serum and BAL fluid from 38 bronchoscopies performed in 9 single lung, 2 bilateral lung, and 4 heart-lung transplant patients were studied. Differential cell counts were made from the BAL fluid. Levels of ECP, myeloperoxidase (MPO), and HYA were used as indirect markers for activation of eosinophils, neutrophils, and fibroblasts, respectively. In addition, levels of circulating ICAM-1, cVCAM-1, and cE-selectin were analyzed. Twenty-two episodes with acute rejection were diagnosed. Of these, 7 were minimal, 13 were mild, and 2 were of moderate character. We found increased levels of ECP and HYA in BAL fluid during mild acute rejection of the allograft. Numbers of eosinophils were also increased. Activation of neutrophils or neutrophil numbers were not significantly increased. Levels of circulating ICAM-1, cVCAM-1, and cE-selectin did not differ between the groups. This retrospective study shows that measurements of ECP and HYA can give information about the inflammatory process present during acute rejection in patients who have undergone lung transplants. Analysis of cCAMS, however, appears to be of limited value as markers for acute rejection.

Acute Disease↗

Serum level of eosinophil cationic protein in patients with chronic cough: relationship to blood eosinophils and airway hyperresponsiveness.

We measured the serum level of eosinophil cationic protein (ECP), determined the blood eosinophil count, and assessed pulmonary function by spirometry and airway responsiveness to methacholine in 80 patients with a cough lasting longer than 3 weeks without an obvious cause. The serum level of ECP was above the cutoff value of 15.7 ng/mL (mean + 2 SD in 105 healthy control adults) in 30 (37.5%) of 80 patients (high-serum-ECP group). The blood eosinophil count was significantly higher in the high-serum-ECP group than in the normal-serum-ECP group (p < 0.01). The cumulative dose of methacholine causing a 35% decrease in respiratory conductance (PD35Grs) was significantly lower in the high-serum-ECP group than in the normal-serum-ECP group (p < 0.001). The serum concentration of ECP was correlated with the blood eosinophil count and the PD35Grs (r = 0.59, p < 0.001 and r = -0.48, p < 0.001, respectively). These findings suggest a possible role for serum level of ECP in management of patients with chronic cough.

Adult↗

Major interference from leukocytes in reverse transcription-PCR identified as neurotoxin ribonuclease from eosinophils: detection of residual chronic myelogenous leukemia from cell lysates by use of an eosinophil-depleted cell preparation.

BACKGROUND: The extraction of RNA from leukocytes for reverse transcription-PCR (RT-PCR) is time-consuming and contributes to variation in analysis of the Philadelphia (Ph1) chromosome of chronic myelogenous leukemia (CML) by RT-PCR. To detect residual CML after bone marrow transplantation, mRNA from at least 10(5) leukocytes should be analyzed, but the RNase activity of the cells precludes simple leukocytes lysis as an alternative to RNA extraction. We sought to identify the main source of RNase activity of leukocytes. METHODS: We used a three-step chromatographic process and amino acid sequence analysis. We selected eosinophil-free granulocytes by using a biotinylated CD16 antibody and selected mononuclear cells by fractionating the leukocytes with a Ficoll-Paque(R) density gradient. RESULTS: Chromatography and amino acid sequencing identified eosinophil-derived neurotoxin (EDN) as the main source of leukocyte RNase. Depletion of eosinophils reduced the EDN content of cell lysates by approximately 90%, allowing a signal from a lysate of 50 K562 Ph1-positive cells mixed with 10(5) CD16(+) granulocytes that was equivalent to 77% of the signal in the absence of leukocytes. A similar lysate with mononuclear cells gave a signal equivalent to 53% of that without mononuclear cells. RNA extraction gave a signal equivalent to only 24% of the leukocyte-free control. CONCLUSION: The depletion of eosinophils during the preparation of leukocyte samples for RT-PCR efficiently reduces the risk of mRNA degradation by ribonucleases, enabling RT-PCR analysis directly from cell lysates with a better signal than can be obtained by RNA extraction.

Amino Acid Sequence↗

Eosinophil counts and urinary eosinophil protein X in children hospitalized for wheezing during the first year of life: prediction of recurrent wheezing.

UNLABELLED: Early identification of wheezing children with an increased risk of recurrent wheezing or subsequent asthma is important. The aim of the study was to determine the role of markers of eosinophil activation, along with other parameters, in the prediction of recurrent wheezing and allergic sensitization in children with early and severe wheezing. We examined 105 children without atopic dermatitis, hospitalized for wheezing during the first year of life. At a 20-mo follow-up, 101 of the children were assessed for the occurrence of recurrent wheezing (at least 3 episodes, including 1 in the previous 6 mo) and allergic sensitization (positive skin-prick test). By univariate analysis, levels of eosinophil counts at the time of hospitalization (p = 0.005, OR = 18.9), age in months (p < 0.0001, OR = 1.5), respiratory syncytial virus (RSV)-negative disease (p < 0.0001, OR = 8.8), parental atopy (p = 0.006, OR = 3.3) and male sex (0.02, OR = 2.7) were all predictive factors for recurrent wheezing at follow-up. With all parameters included in a multiple regression analysis, RSV-negative disease was not a predictive factor for recurrent wheezing. A simple model including eosinophil counts > or = 0.1 x 10(9)/L and age had a predictive accuracy of 79%, with only a 6% chance of a child being wrongly predicted as symptomatic. Urinary protein X (U-EPX) was not a predictive factor for recurrent wheezing. When included in a multiple logistic regression analysis, a level of U-EPX > or = 100 microg/mmol creatinine was the only parameter with a positive predictive value for allergic sensitization (p = 0.007, OR = 18.9), whereas age, parental allergy or parental asthma were not. CONCLUSION: Children with severe wheezing during the first year of life and subsequent recurrent wheezing are characterized by a normal or high eosinophil count in response to viral infections.

Asthma↗