Eosinophilic esophagitis auxiliary diagnosis based on a peptide ligand to eosinophil cationic protein in esophageal mucus of pediatric patients.

Eosinophilic esophagitis (EoE) is a chronic inflammatory condition of the esophagus characterized by increased number of eosinophils. Currently, EoE diagnosis is based on endoscopic procedures for histopathological examination, eosinophils' counting and, often, in clinical practice, the challenge is the differentiation between EoE and gastroesophageal reflux disease (GERD). Our aim was to develop novel peptide ligand to Eosinophil cationic protein (ECP) present in EoE biopsies of patients with potential to be used for detection. We performed a comparative proteomic analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) of esophageal biopsies from pediatric patients with eosinophilic esophagitis, gastroesophageal reflux disease and control individuals. Then, phage display technology was used to select peptides against specific up-regulated protein from EoE patients. Twelve phage clones were selected after three biopanning rounds, and the best phage clone reactivity was evaluated by phage-ELISA assay using esophageal mucus samples from 94 pediatric patients. Mass spectrometry showed that eosinophil cationic protein (ECP) was one of the most up-regulated proteins in EoE patients, which is an eosinophil granule protein usually deposited on tissues to mediate remodeling, but in excess may cause fibrosis and hypertrophy, especially in allergic responses. A highly reactive ECP-ligand peptide (E5) was able to distinguish reactive mucus of EoE patients from GERD and the control individuals by Phage-ELISA, achieving a sensitivity of 84.62%, and a specificity of 82.72%. This is the first study that successfully demonstrated an antibody-like peptide targeting ECP at the esophagus mucus as a useful auxilliary tool for EoE diagnosis with a significant association with atopic disorders and dysphagia.ClinicalTrials.gov no.: NCT03069573.

Eosinophilic Esophagitis Through the Flexible Endoscope and the Lens of a Photographer in the Amazon.

Eosinophilic esophagitis (EoE) is a disorder that has been identified recently, thus knowledge about it, its pathogenesis, and potential etiologies has spread in an era where the medical community and the public are receiving the information and discussing it as it appears in the medical literature. Because physiology, pathology, and pathophysiology are difficult to explain in layman terms, the author has used photographs taken in remote areas of the Amazon to create visual similes within a narrative that brings the scientific and medical concepts of the knowledge on EoE to a level that allows both medical and non-medical persons to grasp and discuss their significance. This set of photographs when presented to audiences has generated interest in the disorder as well as in the Amazon and its natural flora and fauna. The author hopes that this pictorial introduction sets the stage for the multiple novel topics reviewed and presented in this issue.

Cytotoxic Activities of Eosinophil Cationic Protein and Eosinophil-derived Neurotoxin: In Silico Analysis.

BACKGROUND: Eosinophil cationic protein (ECP) and eosinophil derived-neurotoxin (EDN) are homologous ribonuclease (RNAse) A family proteins. The objective of the present study was to in silico characterize ECP and EDN with respect to their cytotoxic activities. MATERIALS AND METHODS: Structural, physicochemical, and conserved domain characterizations were carried-out using open-source software, such as InterProScan, NetOGlyc, NetPhos and Discovery Studio 3.1. RESULTS: The proteins did not have atypical conserved domains. EDN had a greater number of glutamine amino acid residues, whereas ECP had a predominance of arginine. ECP had four possible N-glycosylation, three O-glycosylation and four phosphorylation sites. EDN had five putative N-glycosylation, three phosphorylation and no O-glycosylation sites. CONCLUSION: The greater cationicity of ECP may be related to its higher cytotoxicity and to the fact that the varying post-translational modification profiles can generate functional differences from structural alteration. In vivo and in vitro studies need to be performed in order to confirm these predictions.

Eosinophil cationic protein mRNA expression in children with bronchial asthma.

Studies have shown that eosinophils are closely related to pathogenesis of bronchial asthma. Eosinophils release eosinophil cationic protein (ECP), which plays an important role in infection and allergic reactions. Serum ECP mRNA expression in children with bronchial asthma has not been adequately investigated. We analyzed serum ECP mRNA expression in 63 children with bronchial asthma and 21 healthy children by using reverse-transcriptase polymerase chain reaction to understand the role of ECP in children with bronchial asthma. The children with bronchial asthma were segregated into acute-phase and stable-phase groups, based on the severity of the illness. Serum ECP mRNA expression in children with bronchial asthma (0.375 ± 0.04) was significantly higher than that in healthy controls (0.20 ± 0.02; P < 0.05). Additionally, children in the acute-phase group showed higher ECP mRNA expression level (0.44 ± 0.06) than those in the stable-phase (0.31 ± 0.03) and healthy control groups (0.20 ± 0.02; P < 0.05), while the level in the stable-phase (0.31 ± 0.03) was markedly higher than that in the healthy control group (0.20 ± 0.02; P < 0.05). Detection of serum ECP mRNA expression level has possible applications in the diagnosis and treatment of children with bronchial asthma.

Synergistic interactions of begomoviruses with Sweet potato chlorotic stunt virus (genus Crinivirus) in sweet potato (Ipomoea batatas†L.).

Three hundred and ninety-four sweet potato accessions from Latin America and East Africa were screened by polymerase chain reaction (PCR) for the presence of begomoviruses, and 46 were found to be positive. All were symptomless in sweet potato and generated leaf curling and/or chlorosis in Ipomoea setosa. The five most divergent isolates, based on complete genome sequences, were used to study interactions with Sweet potato chlorotic stunt virus (SPCSV), known to cause synergistic diseases with other viruses. Co-infections led to increased titres of begomoviruses and decreased titres of SPCSV in all cases, although the extent of the changes varied notably between begomovirus isolates. Symptoms of leaf curling only developed temporarily in combination with isolate StV1 and coincided with the presence of the highest begomovirus concentrations in the plant. Small interfering RNA (siRNA) sequence analysis revealed that co-infection of SPCSV with isolate StV1 led to relatively increased siRNA targeting of the central part of the SPCSV genome and a reduction in targeting of the genomic ends, but no changes to the targeting of StV1 relative to single infection of either virus. These changes were not observed in the interaction between SPCSV and the RNA virus Sweet potato feathery mottle virus (genus Potyvirus), implying specific effects of begomoviruses on RNA silencing of SPCSV in dually infected plants. Infection in RNase3-expressing transgenic plants showed that this protein was sufficient to mediate this synergistic interaction with DNA viruses, similar to RNA viruses, but exposed distinct effects on RNA silencing when RNase3 was expressed from its native virus, or constitutively from a transgene, despite a similar pathogenic outcome.

Eosinophil cationic protein: overview of biological and genetic features.

The eosinophil cationic protein (ECP) is a small polypeptide that originates from activated eosinophil granulocytes. A wide range of stimuli has been shown to induce the secretion of ECP. The gene that encodes the human ECP is located on chromosome 14, and the protein shares the overall three-dimensional structure and the RNase active-site residues with other proteins in the RNase A superfamily. Several single-nucleotide polymorphisms in the human ECP gene have been currently described. ECP has many biological functions, including an immunoregulatory function, the regulation of fibroblast activity, and the induction of mucus secretion in the airway. Additionally, the protein is a potent cytotoxic molecule and has the capacity to kill mammalian and nonmammalian cells. The purpose of this article was to review the known biological and genetic characteristics of ECP that contribute to the understanding of this protein's role in the development and progression of a wide variety of diseases.

The role of eosinophils and eosinophil cationic protein in oral cancer: a review.

Eosinophils are multifunctional leukocytes implicated in the pathogenesis of numerous inflammatory processes, such as allergies and parasitic infections. Increased number of these cells has been described in many human cancers, including oral squamous cell carcinoma, and its presence related to favourable as well as unfavourable prognosis. Although the exact role of eosinophils in tumours is not yet defined, the anti-tumour activity of these rare granulocytes has been associated with the release of their cytotoxic proteins, especially the eosinophil cationic protein, a single polypeptide chain with a molecular mass ranging from 15 to 22kDa encoded by the RNSE3 gene located on chromosome 14q11.2. Functional studies have implicated ECP in numerous processes, such as tissue remodelling in allergic inflammation; however its most striking function is the cytotoxic activity. The aim of this review is to summarise the role and functions of eosinophils and their granule-derived products in oral malignant tumours and the clinical value of the tumour-associated tissue eosinophilia for patients' prognosis.

The 434(G>C) polymorphism in the eosinophil cationic protein gene and its association with tissue eosinophilia in oral squamous cell carcinomas.

OBJECTIVE: The aim of this study was to investigate the prevalence of the Eosinophil cationic protein (ECP)-gene polymorphism 434(G>C) in oral squamous cell carcinoma (OSCC) patients and its association with tumor-associated tissue eosinophilia (TATE), demographic, clinical, and microscopic variables. METHODS: The ECP genotypes of 165 healthy individuals and 157 OSCC patients were detected by PCR-RFLP analysis after cleavage of the amplified DNA sequence with enzyme PstI. TATE was obtained by morphometric analysis. Chi-square test or Fisher's exact test was used to analyze the association of ECP-gene polymorphism 434(G>C) with TATE, demographic, clinical, and microscopic variables in OSCC patients. Disease-free survival and overall survival were calculated by the Kaplan-Meier product-limit actuarial method and the comparison of the survival curves were performed using log rank test. RESULTS: Most of healthy individuals (53.33%) and OSCC patients (57.97%) were heterozygous for the ECP 434(G>C) polymorphism. Based on numerical differences, our results showed that OSCC patients with intense TATE and at least one C allele had a higher frequency of bilateral neck dissection, local recurrence, vascular embolization, involved resection margins, and postoperative radiotherapy. No statistically significant differences on survival rates were found in OSCC patients presenting different ECP 434(G>C) genotypes. CONCLUSIONS: These results suggest a tendency towards a poor clinical outcome in OSCC patients with intense TATE and 434GC/CC genotypes, probably due to an ECP genetic variant with altered cytotoxic activity.

Eosinófilos e proteína catiônica eosinofílica em bronquiolite viral aguda / Eosinophil counts and eosinophil cationic protein in acute viral bronchiolitis

Introdução: A bronquiolite viral aguda é uma doença freqüente na infância. Tem sido estudado o papel de marcadores imunológicos e inflamatórios como determinantes da sua gravidade. O objetivo deste estudo foi determinar a contagem de eosinófilos no sangue periférico e a concentração sérica de proteína catiônica eosinofílica em pacientes internados com bronquiolite viral aguda e comparar com diferentes graus de gravidade. Metodologia: A contagem de eosinófilos e a concentração da proteína catiônica eosinofílica foram determinadas em lactentes menores de 1 ano, hospitalizados com quadro clínico de bronquiolite. Foram realizadas comparações do número de eosinófilos e da concentração de proteína catiônica,de acordo com gravidade, sexo e faixa etária. Foram utilizados como critérios de gravidade a saturação em ar ambiente e a prevalência de ventilação mecânica. Resultados: Foram estudados 58 pacientes, com idade média de 2 meses. Dezenove pacientes foram classificados como graves, de acordo com a saturação da hemoglobina, inferior a 90%. Sete pacientes necessitaram ventilação mecânica (12%). A contagem de eosinófilos variou de 0 a 1104, com mediana de 100. O número de eosinófilos foi significativamente menor nos pacientes com saturação mais baixa (p<0.05). A concentração da proteína catiônica eosinofílica variou de 2 a 114 mg/litro, com mediana em 6 mg/l. Não houve correlação com a saturação. Conclusão: Houve associação entre baixo número de eosinófilos e gravidade da bronquiolite medida pela satHb. Estudos adicionais são necessários para elucidar melhor o papel de eosinófilos e seus derivados na determinação da gravidade da bronquiolite.

Introduction: The aim of the present study is to determine peripheral blood eosinophil counts and eosinophil cationic protein (ECP) serum concentration in hospitalized patients with acute viral bronchiolitis and to compare with different degrees of severity. Methodology: Peripheral blood eosinophil counts and eosinophil cationic protein serum concentration was performed on infants under 12 months of age who were hospitalized presenting acute viral bronchiolitis. Associations of eosinophil counts and ECP concentration among groups of patients were performed according to severity, gender and age.Severity was defined by haemoglobin saturation in room air and prevalence of mechanical ventilation. Results: Fifty-eight patients were studied. The mean age was 2 months. Nineteen patients were classified as severe according to haemoglobin saturation, lower than 90%. Seven patients needed mechanical ventilation (12%). Eosinophil counts ranged from 0 to 1104 and the median value was 100. Eosinophil counts were lower in patients with lower saturation (p<0.05). Patients who needed mechanical ventilation also showed lower eosinophil counts. ECP serum concentration ranged from 2 to 114 mg/land the median value was 6 mg/l. ECP was not associated with neither saturation nor mechanical ventilation. Conclusion: A correlation was found between eosinophil counts and severity measured by haemoglobin saturation. There was no correlation between ECP and severity. Additional studies are necessary to better explain the role of eosinophils and its proteins to determine the severity of bronchiolitis.

[Distribution of eosinophils at different stages of hepatic granuloma evolution in mice infected with Schistosoma mansoni]. / A distribuição dos eosinófilos nas diferentes fases de evolução do granuloma hepático em camundongos infectados pelo Schistosoma mansoni.

In the present study, the distribution of eosinophils at different stages of the formation of hepatic granuloma in mice infected with Schistosoma mansoni was evaluated. From the results obtained, we suggest a new classification for the evolution of hepatic granuloma in mice, constructed from the phases described by other authors. In each phase, there is a different pattern of eosinophil distribution. In the exudative-necrotic phase, the eosinophils are concentrated in the periphery and center of the granuloma, and are scarce in the necrotic area; in the productive phase, the eosinophils are dispersed throughout the granuloma; and in the cure due to fibrosis phase, the eosinophils are concentrated in the periphery and center of the granuloma. Eosinophils were found in direct contact with the eggs at all stages of evolution of the granuloma. It was concluded that the dynamics of eosinophils have an important role in forming the granulomatous reaction of the host and in resolving the inflammatory process caused by the parasite egg, as well as adding new data regarding hepatic granuloma classification.

A distribuição dos eosinófilos nas diferentes fases de evolução do granuloma hepático em camundongos infectados pelo Schistosoma mansoni / Distribution of eosinophils at different stages of hepatic granuloma evolution in mice infected with Schistosoma mansoni

No presente estudo, avaliou-se a distribuição dos eosinófilos nas diferentes fases da formação do granuloma hepático de camundongos infectados pelo Schistosoma mansoni. A partir dos resultados obtidos sugerimos uma nova classificação para a evolução do granuloma hepático em camundongos montada a partir de fases descritas por outros autores. Em cada fase há um padrão diferente de distribuição dos eosinófilos. Na fase necrótico-exudativa os eosinófilos encontram-se concentrados na periferia e no centro do granuloma e na área de necrose eles são escassos; na "produtiva" os eosinófilos estão ainda distribuídos de maneira difusa por todo o granuloma; na de cura por fibrose se concentram na periferia e no centro do granuloma. Os eosinófilos estavam em contato direto com os ovos em todos os estágios de evolução dos granulomas. Conclui-se então que a dinâmica dos eosinófilos possui papel importante na formação da reação granulomatosa do hospedeiro e resolução do processo inflamatório causado pelo ovo do parasita, além de acrescentar novos dados na classificação dos granulomas hepáticos.

In the present study, the distribution of eosinophils at different stages of the formation of hepatic granuloma in mice infected with Schistosoma mansoni was evaluated. From the results obtained, we suggest a new classification for the evolution of hepatic granuloma in mice, constructed from the phases described by other authors. In each phase, there is a different pattern of eosinophil distribution. In the exudative-necrotic phase, the eosinophils are concentrated in the periphery and center of the granuloma, and are scarce in the necrotic area; in the productive phase, the eosinophils are dispersed throughout the granuloma; and in the cure due to fibrosis phase, the eosinophils are concentrated in the periphery and center of the granuloma. Eosinophils were found in direct contact with the eggs at all stages of evolution of the granuloma. It was concluded that the dynamics of eosinophils have an important role in forming the granulomatous reaction of the host and in resolving the inflammatory process caused by the parasite egg, as well as adding new data regarding hepatic granuloma classification.

Interrelationship between serum and sputum inflammatory mediators in chronic obstructive pulmonary disease.

Little is known about airway inflammatory markers in chronic obstructive pulmonary disease (COPD). The objective of the present study was to identify and try to correlate pulmonary and peripheral blood inflammatory markers in COPD. In a cross-sectional study on patients with stable COPD, induced sputum and blood samples were collected for the determination of C-reactive protein, eosinophilic cationic protein, serum amyloid A protein, alpha-1 antitrypsin (alpha-1AT), and neutrophil elastase. Twenty-two patients were divided into two groups according to post-bronchodilator forced expiratory volume in the first second (%FEV1): group 1 (N = 12, FEV1 <40%) and group 2 (N = 10, FEV1 > or =40%). An increase in serum elastase, eosinophilic cationic protein and alpha-1AT was observed in serum markers in both groups. Cytology revealed the same total number of cells in groups 1 and 2. There was a significantly higher number of neutrophils in group 1 compared to group 2 (P < 0.05). No difference in eosinophils or macrophages was observed between groups. Serum elastase was positively correlated with serum alpha-1AT (group 1, r = 0.81, P < 0.002 and group 2, r = 0.83, P < 0.17) and negatively correlated with FEV1 (r = -0.85, P < 0.03 and -0.14, P < 0.85, respectively). The results indicate the presence of chronic and persistent pulmonary inflammation in stable patients with COPD. Induced sputum permitted the demonstration of the existence of a subpopulation of cells in which neutrophils predominated. The serum concentration of all inflammatory markers did not correlate with the pulmonary functional impairment.

Interrelationship between serum and sputum inflammatory mediators in chronic obstructive pulmonary disease

Little is known about airway inflammatory markers in chronic obstructive pulmonary disease (COPD). The objective of the present study was to identify and try to correlate pulmonary and peripheral blood inflammatory markers in COPD. In a cross-sectional study on patients with stable COPD, induced sputum and blood samples were collected for the determination of C-reactive protein, eosinophilic cationic protein, serum amyloid A protein, a-1 antitrypsin (a-1AT), and neutrophil elastase. Twenty-two patients were divided into two groups according to post-bronchodilator forced expiratory volume in the first second ( percentFEV1): group 1 (N = 12, FEV1 <40 percent) and group 2 (N = 10, FEV1 ³40 percent). An increase in serum elastase, eosinophilic cationic protein and a-1AT was observed in serum markers in both groups. Cytology revealed the same total number of cells in groups 1 and 2. There was a significantly higher number of neutrophils in group 1 compared to group 2 (P < 0.05). No difference in eosinophils or macrophages was observed between groups. Serum elastase was positively correlated with serum a-1AT (group 1, r = 0.81, P < 0.002 and group 2, r = 0.83, P < 0.17) and negatively correlated with FEV1 (r = -0.85, P < 0.03 and -0.14, P < 0.85, respectively). The results indicate the presence of chronic and persistent pulmonary inflammation in stable patients with COPD. Induced sputum permitted the demonstration of the existence of a subpopulation of cells in which neutrophils predominated. The serum concentration of all inflammatory markers did not correlate with the pulmonary functional impairment.

Níveis séricos de proteína catiônica eosinofílica e contagem de eosinófilos em crianças enteroparasitadas,residentes em área de baixo nível sócio-econômico na cidade de Natal, RN, Brasil / Serum eosinophil cationic protein levels end eosinophil counting in children enteroparasited

A Proteína Catiônica Eosinofílica (ECP) foi purificada de células humanas, pela primeira vez, em 1971 e, após cinco anos, foi identificada como sendo uma proteína granular eosinofílica que possui uma potente toxicidade para uma variedade de helmintos, bactérias e outros microrganismos. O objetivo deste trabalho foi diagnosticar as infecções por enteroparasitas correlacionando com níveis séricos de proteína catiônica eosinofílica e eosinofilia. Foram estudadas 150 crianças de ambos os sexos, com idade variando entre 3 e 6 anos, residentes no mesmo bairro e frequentadoras de creche. A Infecção Parasitária foi investigada por Exames de fezes pelosMétodos de Hoffmann, Pons & Janer e o de Baermann-Morais. Os níveis séricos de proteína catiônica eosinofílica foram determinadospor Fluoroenzimaimunoensaio, utilizando o Kit Unicap (Pharmacia & Upjonh) e a contagem de eosinófilos realizada em esfregaço sanguíneo corado pelo Leishman. Das crianças estudadas 140 (93,3%) apresentaram infecção por enteroparasitas e 10 (6,7%) apresentaram ausência de ovos e larvasde helmintos e cistos de protozoários. Cento e quarenta e oito amostras de soro foram analisadas para determinar os níveis de proteína catiônica eosinofílica e os resultados obtidos mostraram que foi 45,45μg/L a mediana das concentrações observadas e que com relação aos níveis de ECP em crianças parasitadas e não parasitadas por helmintos observou-se que as crianças parasitadas apresentaramconcentrações (MD=52,20μg/L) significantemente mais elevadas do que as não parasitadas (MD=29,70μg/L) e que houve uma correlação positiva entre níveis séricos de ECP e eosinófilos ( p< 0,0001 e r= 0,57).

The eosinophil cationic protein was purified from human cells, for the fist time in 1971, after five years, it was identified as being a eosinophil granular protein that is potencially toxic for helminthics, bacteria and other microorganisms. The purpose of this study was to diagnose the enteroparasitics infections co-related to the serum levels of eosinophil cationic protein and eosinophilia. One hundred and fifty children form both sexes, ages varying from 3 to 6 years old who live in the same neighbourhood and go to the same nursery were observed. The parasitic infection was investigated by faecal examinations using the Hoffmann, Pons & Janer and Baermann-morais methods. The serum cationic protein levels were determined by fluorimmunassay using the unicap kit (Pharmacia & Upjohn) and the counting of eosinophis was made by Leishman- stained smears. From the studied children 140 (93,3%) were infected by enteroparasitics and 10 (6,7%) showed no signs of eggs, helminthics, larva,and protozoarios cysts. One hundred and light samples of serum were analysed to determine the levels of eosinophil cationis protein and the obtaind results have showed that 45,45 ìg/L was the overage of the studied concentrations and that the relationship between the levels of eosinophil cationic protein (ECP) in parasited and non-parasited children was that the parasited children showed significantly hight concentrations (MD=52,20ìg/L) than the non-parasited children (MD=29,70 ìg/L). The results also have showed that there was a positive correlation between the serum levels of ECP and eosinophis ( p<0,0001 and r = 0,57).

Associação do polimorfismo do gene da proteína catiônica eosinofílica com a eosinofilia tecidual associada aos tumores em carcinomas espinocelulares de boca / Association of eosinophil cationic protein gene polymorphism with tumor-associated tissue eosinophilia in oral squamous cell carcinomas

A proteína catiônica eosinofílica (ECP) presente nos grânulos específicos dos eosinófilos apresenta atividade citotóxica, particularmente para células tumorais, entretanto a função exata dos eosinófilos e de seus produtos nas neoplasias malignas continua obscura. O objetivo desse trabalho foi investigar a prevalência do polimorfismo 434(G>C) do gene ECP em pacientes com carcinoma espinocelular (CEC) de boca e sua correlação com a eosinofilia tecidual associada aos tumores (TATE), bem como com as características demográficas, clínicas e microscópicas. O genótipo 434 do gene ECP em 165 pacientes saudáveis e em 157 pacientes com CEC de boca, tratados no Hospital do Câncer A. C. Camargo entre 1984 a 2002, foi detectado pela clivagem da seqüência específica de DNA amplificada com a enzima de restrição PstI e análise dos produtos de clivagem pela eletroforese em gel de agarose. A TATE foi determinada por análise morfométrica. A associação entre os genótipos, a intensidade da TATE e as variáveis demográficas, clínicas e microscópicas foi avaliada pelo teste qui-quadrado ou teste exato de Fisher. As análises das sobrevidas global, livre de doença e específica por câncer foram feitas pelo estimador limite de Kaplan-Meier e a comparação das curvas de sobrevida foi realizada utilizando-se o teste log-rank. Notou-se uma predominância dos indivíduos heterozigotos para o polimorfismo 434(G>C) do gene ECP. Nenhuma diferença estatística significativa foi obtida entre os diferentes genótipos, a intensidade da TATE e as variáveis demográficas, clínicas e microscópicas. Uma maior freqüência de esvaziamento cervical bilateral, recidiva local, embolização vascular, comprometimento das margens cirúrgicas e realização de radioterapia pós-operatória foi observada nos pacientes com CEC de boca, TATE intensa e genótipos 434GC/CC. Não houve correlação estatística significativa entre os diferentes genótipos 434 do gene ECP e as sobrevidas global, livre de doença e específica por câncer. Baseados em nossos resultados, concluímos que houve uma tendência de os pacientes com CEC de boca, intensa eosinofilia tecidual e genótipos 434GC/CC do gene ECP apresentarem uma evolução clínica desfavorável, quando comparados aos indivíduos com genótipo 434GG, provavelmente pela presença de uma variante genética dessa proteína com propriedades citotóxicas alteradas.

Eosinophil cationic protein (ECP), found in secretory granules of human eosinophils, presents cytotoxic activity, particularly against cancer cells. The specific functional role of eosinophils in solid malignant tumors remains unclear. The aim of this study was to investigate the prevalence of the ECP-gene polymorphism 434(G>C) in oral squamous cell carcinoma (OSCC) patients and its association with tumor-associated tissue eosinophilia (TATE), as well as demographic, clinical and microscopic variables. The 434 genotypes in the ECP-gene of 165 healthy individuals and 157 OSCC patients, submitted to surgical treatment at the Hospital A. C. Camargo from 1984 to 2002, were detected by cleavage of the amplified DNA sequence with restriction enzyme PstI and analyses of the cleaved product by agarose gel electrophoresis. TATE, in OSCC, was obtained by morphometric analysis. Chisquare test or Fisher’s exact test was used to analyze the association among ECP-gene polymorphism 434(G>C), TATE, demographic, clinical and microscopic variables. Diseasefree survival and overall survival were calculated by the Kaplan-Meier product-limit actuarial method and the comparison of the survival curves were performed using log rank test. Most of healthy individuals and OSCC patients showed the genotype 434GC. There was no statistical association among 434 genotypes, TATE intensity and demographic, clinical or microscopic variables of OSCC patients. Higher frequency of bilateral neck dissection, local recurrence, vascular embolization, involved resection margins and postoperative radiotherapy was detected in OSCC patients with intense TATE and 434GC/CC genotypes. No statistically significant differences on survival rates were found among 434 genotypes. In conclusion, these results suggest a tendency of worse clinical outcome in OSCC patients with intense TATE and 434GC/CC genotypes, probably due an ECP genetic variant with altered cytotoxic activity.

Eosinophil cationic protein damages protoscoleces in vitro and is present in the hydatid cyst.

Eosinophils are locally recruited during the establishment and chronic phases of cystic hydatidosis. This study provides evidence that eosinophil cationic protein (ECP), one of the major components of eosinophil granules, can damage Echinococcus granulosus protoscoleces (PSC). The toxicity of ECP was investigated in vitro by following parasite viability in the presence of this protein. ECP was found to damage PSC at micromolar concentrations; the effect was blocked by specific antibodies and heparin, and was more severe than the one caused by similar concentrations of RNase A, suggesting that the cationic nature of ECP, and not its ribonuclease activity, is involved in toxicity. This observation may highlight the capacity of eosinophils to control secondary hydatidosis, derived from PSC leakage from a primary cyst. To further assess the relevance of the previous result during infection, the presence of eosinophil proteins was investigated in human hydatid cysts. ECP was found to be strongly associated with the laminated layer of the cyst wall, and present at micromolar concentrations in the hydatid fluid. Overall, these results demonstrate that eosinophils degranulate in vivo at the host-parasite interface, and that the released ECP reaches concentrations that could be harmful for the parasite.

Inflammatory mediators in nasal lavage among school-age children from urban and rural areas in São Paulo, Brazil.

CONTEXT: Some studies have shown that inflammatory processes in the nasal air passages may reflect or affect those in the lower airways. We decided to indirectly assess the inflammatory status of the nasal airways in two groups of children with different sensitization rates to aeroallergens. OBJECTIVE: To compare the inflammatory activity in the nasal airways, through the determination of mediators in nasal lavage fluid in two distinct populations. TYPE OF STUDY: Cross-sectional study. SETTING: Two public elementary schools, one in an urban setting and the other in a rural setting of the State of São Paulo, Brazil. METHODS: Two groups of 40 elementary school children with different sensitization rates to aeroallergens were formed. Samples of nasal lavage fluid were assessed for eosinophil cationic protein (ECP) and tryptase. Non-parametric tests were used for statistical analysis. RESULTS: Significantly higher levels of ECP were observed among students living in the urban area than those in the rural area (p < 0.05). No significant difference in the tryptase levels was observed. Also, the urban children who were sensitized to aeroallergens presented higher levels of ECP in nasal mucosa than the non-sensitized children, while this difference was not observed among the rural children. DISCUSSION: The lack of mast cell activity and increased eosinophil degranulation revealed a chronic inflammatory state in the nasal air passages. The higher eosinophil activity in the urban area, coinciding with higher sensitization to aeroallergens, suggests that there must be some factors in the urban area that can modulate airway inflammation by influencing the activation of inflammatory cells. CONCLUSION: Our findings showed that there was no difference in the concentrations of tryptase in nasal lavage fluids between the two studied groups. However, the children from the urban area presented with higher concentrations of eosinophil cationic protein than did those from the rural area. Also, the urban children who were sensitized to aeroallergens presented with greater concentrations of eosinophil cationic protein in nasal mucosa than the non-sensitized children, while this difference was not observed among the rural children.

Small bowel of patients with asthma and allergic rhinitis: absence of inflammation despite the presence of major cellular components of allergic inflammation.

Eosinophils participate in the pathogenesis of inflammatory diseases of the respiratory tract and the gut. We investigated the constitutive presence of eosinophils and mononuclear cells in the macroscopically normal duodenal mucosa of patients with asthma and allergic rhinitis. Macroscopically normal duodenal specimens were obtained at routine endoscopy for upper gastrointestinal symptoms from 16 patients with asthma and 13 patients with allergic rhinitis. Twelve nonatopic patients with irritable bowel syndrome were studied as controls. Specimens were analyzed by immunohistochemistry using a panel of antibodies to human eosinophil cationic protein clone EG1 (EG1) and clone EG2 (EG2), anti-human interleukin (anti-hIL)-5, anti-hIL-4, anti-CD4, and anti-CD68. Significantly increased numbers of eosinophils stained with EG1 and EG2 were found in the duodenum of patients with asthma and allergic rhinitis compared with controls. IL-5+ cells and IL-4+ cells were detected in significantly increased numbers in the duodenal mucosa of patients with asthma and rhinitis compared with controls. Mononuclear cells expressing CD4 (helper T cells) and CD68 (macrophages) also were significantly increased in the duodenal mucosa of asthma and rhinitis compared with controls. Accumulation of eosinophils in conjunction with IL-4+ cells and IL-5+ cells in the noninflamed duodenal mucosa may reflect a predominant T helper cell subset 2 systemic immune response in patients with asthma and allergic rhinitis. The absence of intestinal inflammation despite the marked presence of cells implicated in the allergic inflammation suggests that local mechanisms might determine the state of nonresponsiveness in the gut mucosa of patients with asthma and allergic rhinitis.

Atividade imuno-inflamatória de ovalbumina: papel de células e mediadores do sistema imune intestinal na migração de eosinófilos / Immuno-inflammatory activity of ovalbumin: role of the intestinal cells and immune mediators in the migration of eosinophils

Ratos são comumente usados em modelos de anafilaxia, principalmente em anafilaxia intestinal. Os mecanismos de hipersensibilidade são complexos e não estão completamente esclarecidos. Os objetivos deste estudo foram: a) investigar o efeito pró-inflamatório da ovalbumina no modelo de edema de pata em ratos sensibilizados; b) identificar os mediadores envolvidos nesta imuno-inflamação e, c) investigar o envolvimento de células do sistema imune intestinal e seus mediadores na fisiopatologia da hipersensibilidade intestinal induzida por ovalbumina (ova.). O edema de pata induzido por ova e sua modulação farmacológica foi estudada. Ratos Wistar previamente sensibilizados (14 a 18 dias) com ova (30mg, i.p.) ou pseudo-sensibilizados com hidróxido de alumínio (controle). Os volumes das patas foram mensurados antes dos estímulos antigênicos e 1, 2, 3 e 4 horas após a injeção intraplantar de ova (10 μg/pata). O pré-tratamento s.c. com dexametasona, difenidramina, ciproheptadina, clorpromazina, ou metisergida inibiram significativamente (p < 0,05) o edema de pata. NDGAm, indometacina, MK-886, WEB 2086, cetotifeno e meclizina não inibiram este edema. A talidomida e pentoxifilina foram ineficazes no controle do edema induzido por ovalbumina...