Approximately two billion people are infected with helminths worldwide. In order to develop a vaccine against these pathogens, more needs to be known about the immune response to helminths. Eosinophils are important for resistance to some helminth species and their recruitment to infected tissues, attachment to parasites and degranulation may all be critical processes for immunity. Complement may contribute to these processes via generation of chemotactic factors (C3a and C5a) or opsonisation of the parasite with C3b/iC3b. The importance of complement during helminth infection is unclear, though complement does promote leukocyte-mediated killing of several helminth species in vitro. The aim of the present study was to investigate the role of complement in immunity of mice to Nippostrongylus brasiliensis, with a focus on whether complement facilitates eosinophildependent resistance to this parasite. A new fluorescence-based method for quantifying in vitro complement deposition and leukocyte adherence on N. brasiliensis was developed. C3 from human serum was deposited on infective-stage L3 via the classical or lectin complement pathways. In contrast, the alternative complement pathway mediated binding of mouse C3 and eosinophil-rich mouse peritoneal leukocytes to L3. Interestingly, the ability of complement and leukocytes to bind to the parasite changed as it matured. Larvae recovered from the skin 30 min post-injection (p.i.) were coated with C3, however those harvested 150 min p.i. exhibited reduced C3 binding capacity. Binding of C3 and eosinophils to larvae recovered from the lungs 24-48 h p.i. (L4) was also diminished compared to that seen on L3. Adult intestinal worms bound C3 and leukocytes only when treated ex vivo with serum and cells. Mice lacking in classical (C1q-deficient), alternative (factor B-deficient) or all complement pathways (C3-deficient) were then employed to determine if complement was important for resistance of mice to N. brasiliensis. IL-5 Tg mice deficient in individual complement genes were generated to assess whether complement contributed to eosinophildependent resistance to the parasite. Factor B deficient mice exhibited impaired C3 deposition on larvae, eosinophil recruitment, eosinophil degranulation and larval aggregation in the skin 30 min p.i. Eosinophil recruitment was similarly abolished by treatment of mice with the C5aR inhibitor PMX53. However at 150 min p.i., larval aggregation, eosinophil and neutrophil recruitment, leukocyte adherence and eosinophil degranulation were largely complement-independent. Ablation of factor B or C3 caused minor but significant increases in lung-larval burden during primary, but not in secondary, infections. Critically, a lack of C3 or factor B in IL-5 Tg mice failed to greatly impair the strong innate anti-parasite resistance typical of these animals, suggesting that eosinophils can provide immunity to N. brasiliensis infection in the absence of complement. This was unexpected, given the evidence from this and previous studies which suggested that in vitro, complement is important for promoting eosinophil-dependent killing of N. brasiliensis and other helminth species. The mechanism(s) by which eosinophils kill N. brasiliensis remain unknown, but may involve the coordination of the complement system with complement-independent factors that act in the early stages of infection. Critically, the influence of complement is limited, because soon after entry into the host, the parasite develops the ability to resist complement activation. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1311182 / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Science, 2008

Asthma is an inflammatory airways disease affecting over 339 million people of all ages worldwide. More than 10% of asthmatics have uncontrolled severe disease which is insensitive to high doses of oral corticosteroid treatment. Type 2 innate lymphoid cells (ILC2) are pro-inflammatory lymphomononuclear cells proposed as critical drivers of eosinophilic inflammatory disease of the upper and lower airways. Controlling this activity may provide novel therapies for asthma. This thesis aimed to investigate factors that affect the local activation and expansion of ILC2 in the airways including anti-inflammatory medications such as (i) corticosteroids, (ii) neuro-immune regulation of ILC2, and (iii) effect of locally generated cytokines on ILC phenotypes and the relationship to the ongoing airway inflammatory profile. We firstly investigated the effect of intranasal corticosteroids on activation levels of ILC2 in the upper airway of allergic rhinitics with mild asthma following controlled nasal allergen challenge (Chapter 2). Following pre-treatment with intranasal corticosteroid there was an attenuation in the allergen-induced increase in total ILC2 and IL-5/13+ ILC2 in the nasal mucosa. In addition, HLA-DR expression on ILC2 in the nasal mucosa was down-regulated. Overnight culture with IL-2, TSLP or IFN-γ up-regulated HLA-DR expression on ILC2, in vitro; an effect that is inhibited in the presence of corticosteroids. Attenuation of HLA-DR expression by ILC2 may be an additional mechanism by which corticosteroids modulate adaptive immune responses in the airways. We have previously reported that lung ILC2 are activated within 7h following allergen-inhalation challenge. Since airway mucosal tissue is highly innervated, we investigated whether neuroimmune interactions may trigger early and rapid host immune responses (Chapter 3). In a diluent-controlled allergen-inhalation challenge cross-over study, where mild asthmatics developed early and late bronchoconstrictor responses with sputum eosinophilia (>3%), NMUR1, a receptor for the neuropeptide, neuromedin-U, was up-regulated on sputum ILC2 in 7h post allergen challenge. This was associated with increased expression of IL-5/IL-13 by sputum ILC2 post-allergen and following in vitro culture. ILC2 activation was mediated through a MAPK/PI3 kinase dependent-signaling pathway that was attenuated in the presence of dexamethasone. Co-culture with IL-33 and TSLP, in vitro up-regulated NMUR1 expression on ILC2 at the protein and transcriptomic level which was attenuated by dexamethasone. The close interplay between neuropeptide signalling and tissue-derived alarmin cytokines may be important interactions for rapid ILC2 activation in airway inflammatory responses in asthma. We have reported increased ILC2 with the highest level of IL-5/13+ ILC2 in the airways of severe asthma with uncontrolled eosinophilia (>3%). The prevalence and phenotypic analyses of innate lymphoid cells subsets in severe asthma with neutrophilic or mixed granulocytic airway inflammatory endotypes remains unclear and was investigated in Chapter 4. Sputum ILC3 were most abundant in severe asthma with neutrophilic airway inflammation where IL-17A+ ILC3 correlated with airway neutrophilia. ILC2 were predominant in severe asthma with airway eosinophilia. Importantly, we identified an intermediate ILC2 phenotype displaying ILC3-like markers (c-kit and IL-17A) in severe asthma with neutrophilic and mixed granulocytic airway inflammation. Inflammasome related cytokines, IL-1β and IL-18 were significantly increased in the airways of these patients. At both proteomic and transcriptomic levels, flow sort-purified ILC2 trans-differentiated to the intermediate phenotype when co-cultured with IL-1β+IL-18. Blocking inflammasome-related cytokines may control T2-low severe asthma exacerbations. Collectively, the findings of this thesis highlight the role of corticosteroids, neuropeptides and airway inflammasome related cytokines as modulators of ILC fate and activity in asthma. / Thesis / Doctor of Philosophy (PhD) / Asthma is a disease of the airways that makes breathing difficult. About 10% of asthma patients have uncontrolled severe symptoms despite treatment with high doses of corticosteroids which imposes many unwanted side effects. Investigating processes that worsen the disease will help to discover new treatments for asthma. Type 2 innate lymphoid cells (ILC2) are novel cells that produce large quantities of factors which attract and activate effector cells to the lungs which in turn make breathing difficult. This thesis investigated whether controlling ILC2 activity reduces asthma symptoms by studying i) responsiveness of ILC2 to corticosteroids using a controlled allergen exposure through the nose in people with allergic rhinitis and mild asthma, ii) the role of airway nerves and mediators on ILC2 activation, and iii) the ability of signals produced by the lungs to impact factors released by ILC2 and the relationship to effector cells found in the airways of severe asthma. Overall, ILC2 activation can be modulated by corticosteroids, nerve derived factors and lung tissue derived cytokines, and this is associated with changes in the number and type of effector cells in the lungs.

No description available.

Surfactant proteins, SP-A and SP-D, are collagen-containing calcium-dependent (C-type) lectins, called, collectins. Their primary structure has four regions: a cysteine-linked N- terminal region involved in multimerization, a collagen region composed of Gly-X-Y repeats, coiled-coil neck region, and the C-terminal carbohydrate recognition domains (CRD) or C-type lectin domain. SP-A looks like a bouquet, while SP-D is a cruciform- like structure, with four arms of equal length. SP-A and SP-D have been shown to act as innate immune molecules at pulmonary as well as extra-pulmonary sites by binding to pathogens, allergens and apoptotic/necrotic cells via their CRD region. SP-A and SP-D can induce pathogen neutralization and enhanced phagocytosis. In addition, SP-A and SP-D can interact via CRDs with allergens and dampen allergic reaction in vitro and in vivo. This thesis examines in vitro interaction of a recombinant fragment of human SP-D containing neck and CRD regions (rhSP-D) with IAV and Aspergillus fumigatus, in addition to characterizing a dichotomy of the effects of SP-A and SP-D on dendritic cells in an attempt to explain how SP-A and SP-D modulate DC functions differentially. Experiments involving interaction of rhSP-D with IAV pandemic strain show that it can be a restrictive factor against the virus, in addition to modulating immune response by a macrophage cell line. The rhSP-D can have anti-A. fumigatus effect directly and indirectly in the context of pathogen as well as allergen. A comparison has been made between two recombinant fragments of SP-D that have been expressed with and without 8 Gly-X-Y repeats for their fungistatic properties. The effects of SP-A and SP-D on cultured DC maturation, and effector cytokine and proliferative response of co-cultured cells have also been examined in vitro.

Esofagite Eosinofílica é uma doença inflamatória crônica restrita ao esôfago e imune mediada por antígenos. Sua prevalência descrita varia desde 0,4%, numa população geral, até 15% em pacientes com sintomas de disfagia. Já se conhece sua associação com doenças atópicas, anafilaxia e alergia alimentar, sendo o leite de vaca um dos principais alimentos envolvidos. Existem relatos recentes de casos em que pacientes foram diagnosticados com esofagite eosinofílica após serem submetidos à imunoterapia oral com o alimento causador de sua alergia alimentar mediada por IgE. Porém, em nenhum destes casos foi avaliado previamente se os mesmos pacientes já não apresentavam eosinofilia esofágica latente e/ou sintomas subjetivos sugestivos da doença. Considerando que, atualmente, um dos tratamentos mais promissores para alergia alimentar é a imunoterapia oral, justificou-se a necessidade de entender se esofagite eosinofílica seria de fato uma complicação do tratamento, ou se seria uma condição pré ou coexistente. Portanto, o objetivo deste trabalho foi avaliar a frequência de eosinofilia esofágica em pacientes com anafilaxia à proteína do leite de vaca. Foram analisados 89 pacientes matriculados no ambulatório de alergia alimentar do HC-FMUSP, com mediana de idade de 8 anos e que apresentavam anafilaxia ao leite de vaca. Todos foram submetidos à endoscopia digestiva alta com biópsias de esôfago, estomago e duodeno. Dados demográficos, comorbidades atópicas, uso de medicações e sintomas gastrointestinais foram analisados e comparados. A frequência de eosinofilia esofágica foi de 38,2% (34 de 89 pacientes). Em 15 dos 34 pacientes com eosinofilia esofágica, foi completada a investigação para esofagite eosinofílica com uso de inibidor de bomba de prótons em dose plena por 8 semanas antes de uma segunda endoscopia. Identificou-se, portanto, cinco pacientes (7,1%) com eosinofilia esofágica responsiva a inibidor de bomba de prótons e 10 pacientes com esofagite eosinofílica (14,2%). No grupo total de pacientes com eosinofilia esofágica (n=34) encontrou-se 29,4% de pacientes com quadro clínico gastrointestinal ausente; 23,5% oligossintomáticos, e apenas 47% com sintomas sugestivos de disfunção esofágica e, destes últimos, nem todos apresentavam sintomas esofágicos persistentes. Pode-se concluir que a frequência de esofagite eosinofílica descrita no grupo estudado foi significativamente superior à estimada na população geral e uma das mais altas descritas em grupos de pacientes com fatores de risco específicos. Também foi observada uma grande parcela de pacientes com eosinofilia esofágica, sendo muitos assintomáticos ou oligossintomáticos, surgindo o questionamento se esta não seria uma doença latente, de início precoce, insidioso e não relacionada diretamente como complicação de tratamentos atuais / Eosinophilic esophagitis is a chronic inflammatory disease, which occurs in the esophagus and is immune mediated by antigens. Its observed prevalence varies between 0.4% in the general population to 15% in patients with dysphagia. Its association with atopic diseases, anaphylaxis and food allergy has already been recognized. Cow\'s milk is one of the main food sources involved. There are recent reports of cases in which patients were diagnosed with eosinophilic esophagitis after being submitted to oral immunotherapy with the food that causes the IgE mediated allergy. However, in none of these cases was it previously determined if the same patients did not already present latent esophageal eosinophilia and/or subjective symptoms suggestive of the disease. Considering that, currently, one of the most promising treatment for food allergy is oral immunotherapy, the need to understand if eosinophilic esophagitis could be a treatment complication, or if it is a coexistent or preexistent condition, is justified. Therefore, the objective of this study was to evaluate esophageal eosinophilia frequency in patients with anaphylaxis to cow\'s milk protein. We analyzed eighty-nine patients registered in the Food Allergy Unit of the HCFMUSP, with a median age of 8 years, who presented cow\'s milk anaphylaxis. All of them were submitted to digestive endoscopy as well as esophagus, stomach, and duodenum biopsies. We also analyzed and compared demographic data, atopic comorbidities, use of medication, and gastrointestinal symptoms. The frequency of esophageal eosinophilia was 38.2% (34 of 89 patients). In 15 of the 34 patients with esophageal eosinophilia, full investigation for the disease was carried out using a proton pump inhibitor at full dose for eight weeks prior to a second endoscopy. From this, five patients (7.1%) had the proton pump inhibitor-responsive esophageal eosinophilia phenotype, and ten patients were diagnosed with eosinophilic esophagitis (14.2%). In the whole group of patients with esophageal eosinophilia (n = 34), it was found 29.4% of patients with an absent gastrointestinal clinical condition, 23.5% were oligosymptomatic, and only 47% had symptoms suggestive of esophagic dysfunction. Of these, not all presented persistent esophagic symptoms. It is possible to conclude that the frequency of eosinophilic esophagitis observed in this group was significantly higher than the estimated for the general population, and one of the highest observed in groups of patients with specific risk factors. A large portion of patients with esophageal eosinophilia were oligosymptomatic or asymptomatic, raising the question if this would not in fact be a latent disease, with a precocious beginning, insidious and not directly related to current treatments complications

Estudamos o efeito do tratamento com salbutamol em dois regimes: diário (DS) e administrado a intervalos de 96 horas (IS) em camundongos balb/c sensibilizados com injeções intraperitoneais de uma solução de ovoalbumina (OVA) adsorvida em hidróxido de alumínio, e desafiada com inalações de ovoalbumina a 1%. O grupo controle SAL recebeu injeções i.p. de salina e desafios inalatórios de sallina. A partir do 34o dia, os animais OVA foram tratados com salbutamol via inalatória 10 mg/ml durante 15 minutos nos dois regimes descritos. Os animais foram sacrificados no 60o dia, que corresponde a 48 horas após o último desafio antigênico. Após os camundongos serem anestesiados com pentobarbital sódico via i.p., eles foram traqueostomizados e entubados e sacrificados com secção da Aorta abdominal. Então, procedeu-se com a coleta do lavado broncoalveolar para a quantificação de leucócitos. Coletamos os tecidos pulmonares para a avaliação do processo inflamatório por quantificação de células linfomononucleares (LMN) e eosinófilos EPO+, essa última com marcação citoquímica. Além disso, estudamos a influência do tratamento adrenérgico sobre o IgE anafilático. O modelo de inflamação (grupo OVA) produziu significativo aumento do número de células totais, de eosinófilos e de neutrófilos observados na avaliação de lavado broncoalveolar. Além disso, houve nesse grupo processo inflamatório na parede de vias aéreas, caracterizada por um infiltrado linfomononuclear e com presença de eosinófilos. O nosso processo de indução de inflamação também recrutou eosinófilos para o septo alveolar. O tratamento com salbutamol diário produziu uma queda significativa do processo inflamatório no BAL, principalmente de neutrófilos e eosinófilos, enquanto que o tratamento intermitente produziu redução significativa apenas de neutrófilos. O tratamento com salbutamol a cada 96 horas (IS) promoveu uma queda significativa de células LMN quantificadas no septo alveolar, mas não atingindo valores do grupo salina (NS). Ambos os tratamentos com salbutamol produziu redução significativa de células EPO+ no parênquima pulmonar (P < 0,05). Apesar das alterações no processo celular, o salbutamol não influenciou na expressão de anticorpos IgE anafiláticos a OVA. Assim, podemos concluir que o salbutamol apresenta atividade imunomoduladora, observada por redução de eosinófilos no BAL e no parênquima pulmonar, apesar de não atingir valores semelhantes aos animais do grupo salina / We studied the effects of salbutamol treatment in two regimen: diary (DS) and at interval of 96 hours (IS) in ovalbumin sensitized (OVA) balb/c mice. The control group (NS) received i.p. injections and aerosol challenge with normal saline. Starting at day 34 the OVA animals were treated with 10mg/ml salbutamol by inhalation during 15 minutes per day in both regimen: DS and IS. The mice were sacrificed at day 60 that corresponded the fourthly eight hours after last OVA and/or salbutamol exposure. At experimental day, mice were anesthetized with i.p. injection of sodium pentobarbital, tracheostomized, entubed and the abdominal aorta sectioned. We followed with collecting of bronchoalveolar lavage (BAL) and lungs to histopathology studies. In the BAL, total cells and differential leukocytes were quantified, while in the lung sections, the EPO+ and LMN in airways wall and parenchyma septa were evaluated. Also, we sampled the blood to evaluate the effects of salbutamol on anaphylactic IgE antibodies expression. The inflammatory model (OVA animals) produced a significant increase of BAL total cells, BAL eosinophils and neutrophils, and LMN cells and EPO+ eosinophils in the airways and in the parenchyma. Diary salbutamol treatment decrease significantly BAL eosinophils and neutrophils, while the IS group showed a diminution of BAL neutrophils and LMN cells in the alveolar septum. Both salbutamol treatments produced significant decline of EPO+ cells in the lung parenchyma. Despite the changes in the cellular patterns, the salbutamol did not affect the IgE antibodies expression. So, we can concluded that salbutamol present an immunomodulatory activity observed by reduction of eosinophils in the BAL and lung parenchyma, but did not achieve the values of saline control group

In chronic inflammatory diseases like asthma or rheumatoid arthritis, erroneous and exaggerated accumulation of leukocytes in a tissue inadvertently causes the body harm. Several efficient anti-inflammatory drugs exist, for example corticosteroids and cyclo-oxygenase inhibitors. However, these drugs have potent and diverse effects and often act by inhibiting events subsequent to initiation of the inflammatory response, leading to more or less severe side-effects, especially when used in high doses for long periods of time. For this reason, strategies aimed at early inhibition of recruitment and activation of leukocytes have been suggested as safer and more specific approaches to reduce inflammation. Leukocyte adhesion to activated endothelium is a prerequisite to the following activation and extravasation, and takes place in the initial phase of inflammation. By using a model that allows leukocytes to adhere to tumour necrosis factor (TNF)-activated endothelial cells, thus mimicking aspects of an inflammatory reaction, we found that uridine, 4-thiouridine and isomaltitol could all reduce adhesion. This suggested that they may have anti-inflammatory potential. We therefore tried the three substances in a Sephadex-induced lung inflammation model and found that uridine and 4-thiouridine have several anti-inflammatory effects, such as being able to reduce leukocyte accumulation, decrease TNF protein levels and partly inhibit the oedema induced by Sephadex. Isomaltitol turned out to have immunomodulating, rather than anti-inflammatory, effects, which could be of interest in diseases where inadequate inflammatory responses are a problem.

Esofagite Eosinofílica é uma doença inflamatória crônica restrita ao esôfago e imune mediada por antígenos. Sua prevalência descrita varia desde 0,4%, numa população geral, até 15% em pacientes com sintomas de disfagia. Já se conhece sua associação com doenças atópicas, anafilaxia e alergia alimentar, sendo o leite de vaca um dos principais alimentos envolvidos. Existem relatos recentes de casos em que pacientes foram diagnosticados com esofagite eosinofílica após serem submetidos à imunoterapia oral com o alimento causador de sua alergia alimentar mediada por IgE. Porém, em nenhum destes casos foi avaliado previamente se os mesmos pacientes já não apresentavam eosinofilia esofágica latente e/ou sintomas subjetivos sugestivos da doença. Considerando que, atualmente, um dos tratamentos mais promissores para alergia alimentar é a imunoterapia oral, justificou-se a necessidade de entender se esofagite eosinofílica seria de fato uma complicação do tratamento, ou se seria uma condição pré ou coexistente. Portanto, o objetivo deste trabalho foi avaliar a frequência de eosinofilia esofágica em pacientes com anafilaxia à proteína do leite de vaca. Foram analisados 89 pacientes matriculados no ambulatório de alergia alimentar do HC-FMUSP, com mediana de idade de 8 anos e que apresentavam anafilaxia ao leite de vaca. Todos foram submetidos à endoscopia digestiva alta com biópsias de esôfago, estomago e duodeno. Dados demográficos, comorbidades atópicas, uso de medicações e sintomas gastrointestinais foram analisados e comparados. A frequência de eosinofilia esofágica foi de 38,2% (34 de 89 pacientes). Em 15 dos 34 pacientes com eosinofilia esofágica, foi completada a investigação para esofagite eosinofílica com uso de inibidor de bomba de prótons em dose plena por 8 semanas antes de uma segunda endoscopia. Identificou-se, portanto, cinco pacientes (7,1%) com eosinofilia esofágica responsiva a inibidor de bomba de prótons e 10 pacientes com esofagite eosinofílica (14,2%). No grupo total de pacientes com eosinofilia esofágica (n=34) encontrou-se 29,4% de pacientes com quadro clínico gastrointestinal ausente; 23,5% oligossintomáticos, e apenas 47% com sintomas sugestivos de disfunção esofágica e, destes últimos, nem todos apresentavam sintomas esofágicos persistentes. Pode-se concluir que a frequência de esofagite eosinofílica descrita no grupo estudado foi significativamente superior à estimada na população geral e uma das mais altas descritas em grupos de pacientes com fatores de risco específicos. Também foi observada uma grande parcela de pacientes com eosinofilia esofágica, sendo muitos assintomáticos ou oligossintomáticos, surgindo o questionamento se esta não seria uma doença latente, de início precoce, insidioso e não relacionada diretamente como complicação de tratamentos atuais / Eosinophilic esophagitis is a chronic inflammatory disease, which occurs in the esophagus and is immune mediated by antigens. Its observed prevalence varies between 0.4% in the general population to 15% in patients with dysphagia. Its association with atopic diseases, anaphylaxis and food allergy has already been recognized. Cow\'s milk is one of the main food sources involved. There are recent reports of cases in which patients were diagnosed with eosinophilic esophagitis after being submitted to oral immunotherapy with the food that causes the IgE mediated allergy. However, in none of these cases was it previously determined if the same patients did not already present latent esophageal eosinophilia and/or subjective symptoms suggestive of the disease. Considering that, currently, one of the most promising treatment for food allergy is oral immunotherapy, the need to understand if eosinophilic esophagitis could be a treatment complication, or if it is a coexistent or preexistent condition, is justified. Therefore, the objective of this study was to evaluate esophageal eosinophilia frequency in patients with anaphylaxis to cow\'s milk protein. We analyzed eighty-nine patients registered in the Food Allergy Unit of the HCFMUSP, with a median age of 8 years, who presented cow\'s milk anaphylaxis. All of them were submitted to digestive endoscopy as well as esophagus, stomach, and duodenum biopsies. We also analyzed and compared demographic data, atopic comorbidities, use of medication, and gastrointestinal symptoms. The frequency of esophageal eosinophilia was 38.2% (34 of 89 patients). In 15 of the 34 patients with esophageal eosinophilia, full investigation for the disease was carried out using a proton pump inhibitor at full dose for eight weeks prior to a second endoscopy. From this, five patients (7.1%) had the proton pump inhibitor-responsive esophageal eosinophilia phenotype, and ten patients were diagnosed with eosinophilic esophagitis (14.2%). In the whole group of patients with esophageal eosinophilia (n = 34), it was found 29.4% of patients with an absent gastrointestinal clinical condition, 23.5% were oligosymptomatic, and only 47% had symptoms suggestive of esophagic dysfunction. Of these, not all presented persistent esophagic symptoms. It is possible to conclude that the frequency of eosinophilic esophagitis observed in this group was significantly higher than the estimated for the general population, and one of the highest observed in groups of patients with specific risk factors. A large portion of patients with esophageal eosinophilia were oligosymptomatic or asymptomatic, raising the question if this would not in fact be a latent disease, with a precocious beginning, insidious and not directly related to current treatments complications

Estudamos o efeito do tratamento com salbutamol em dois regimes: diário (DS) e administrado a intervalos de 96 horas (IS) em camundongos balb/c sensibilizados com injeções intraperitoneais de uma solução de ovoalbumina (OVA) adsorvida em hidróxido de alumínio, e desafiada com inalações de ovoalbumina a 1%. O grupo controle SAL recebeu injeções i.p. de salina e desafios inalatórios de sallina. A partir do 34o dia, os animais OVA foram tratados com salbutamol via inalatória 10 mg/ml durante 15 minutos nos dois regimes descritos. Os animais foram sacrificados no 60o dia, que corresponde a 48 horas após o último desafio antigênico. Após os camundongos serem anestesiados com pentobarbital sódico via i.p., eles foram traqueostomizados e entubados e sacrificados com secção da Aorta abdominal. Então, procedeu-se com a coleta do lavado broncoalveolar para a quantificação de leucócitos. Coletamos os tecidos pulmonares para a avaliação do processo inflamatório por quantificação de células linfomononucleares (LMN) e eosinófilos EPO+, essa última com marcação citoquímica. Além disso, estudamos a influência do tratamento adrenérgico sobre o IgE anafilático. O modelo de inflamação (grupo OVA) produziu significativo aumento do número de células totais, de eosinófilos e de neutrófilos observados na avaliação de lavado broncoalveolar. Além disso, houve nesse grupo processo inflamatório na parede de vias aéreas, caracterizada por um infiltrado linfomononuclear e com presença de eosinófilos. O nosso processo de indução de inflamação também recrutou eosinófilos para o septo alveolar. O tratamento com salbutamol diário produziu uma queda significativa do processo inflamatório no BAL, principalmente de neutrófilos e eosinófilos, enquanto que o tratamento intermitente produziu redução significativa apenas de neutrófilos. O tratamento com salbutamol a cada 96 horas (IS) promoveu uma queda significativa de células LMN quantificadas no septo alveolar, mas não atingindo valores do grupo salina (NS). Ambos os tratamentos com salbutamol produziu redução significativa de células EPO+ no parênquima pulmonar (P < 0,05). Apesar das alterações no processo celular, o salbutamol não influenciou na expressão de anticorpos IgE anafiláticos a OVA. Assim, podemos concluir que o salbutamol apresenta atividade imunomoduladora, observada por redução de eosinófilos no BAL e no parênquima pulmonar, apesar de não atingir valores semelhantes aos animais do grupo salina / We studied the effects of salbutamol treatment in two regimen: diary (DS) and at interval of 96 hours (IS) in ovalbumin sensitized (OVA) balb/c mice. The control group (NS) received i.p. injections and aerosol challenge with normal saline. Starting at day 34 the OVA animals were treated with 10mg/ml salbutamol by inhalation during 15 minutes per day in both regimen: DS and IS. The mice were sacrificed at day 60 that corresponded the fourthly eight hours after last OVA and/or salbutamol exposure. At experimental day, mice were anesthetized with i.p. injection of sodium pentobarbital, tracheostomized, entubed and the abdominal aorta sectioned. We followed with collecting of bronchoalveolar lavage (BAL) and lungs to histopathology studies. In the BAL, total cells and differential leukocytes were quantified, while in the lung sections, the EPO+ and LMN in airways wall and parenchyma septa were evaluated. Also, we sampled the blood to evaluate the effects of salbutamol on anaphylactic IgE antibodies expression. The inflammatory model (OVA animals) produced a significant increase of BAL total cells, BAL eosinophils and neutrophils, and LMN cells and EPO+ eosinophils in the airways and in the parenchyma. Diary salbutamol treatment decrease significantly BAL eosinophils and neutrophils, while the IS group showed a diminution of BAL neutrophils and LMN cells in the alveolar septum. Both salbutamol treatments produced significant decline of EPO+ cells in the lung parenchyma. Despite the changes in the cellular patterns, the salbutamol did not affect the IgE antibodies expression. So, we can concluded that salbutamol present an immunomodulatory activity observed by reduction of eosinophils in the BAL and lung parenchyma, but did not achieve the values of saline control group

Introdução: O estudo dos fatores clínicos associados ao prognóstico da rinossinusite crônica (rsc), seja associada à polipose nasossinusal (RSCcPN) ou não (RSCsPN), ainda é pouco abordado a longo prazo. Objetivo: Avaliar pacientes submetidos à ESS (cirurgia endoscópica nasal, do inglês endoscopic sinus surgery) para o tratamento de RSC no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo, entre 1996 e 2006, e correlacionar a recidiva em longo prazo com parâmetros como a extensão da doença, atopia, tabagismo, asma, eosinofilia e doença respiratória exacerbada pela aspirina (DREA). Métodos: Duzentos e um pacientes foram seguidos por um período médio de 12 anos. Os dados clínicos foram levantados, assim como exames de endoscopia nasal, Tomografia Computadorizada (TC), exames séricos, prick test e prova de função pulmonar. O tempo de seguimento pós-operatório foi analisado, sendo considerado fator de mau prognóstico a indicação de novo procedimento cirúrgico. Foi realizada comparação entre os fatores pela curva de Kaplan-Meyer, e pós-teste de Log-rank. Resultados e Discussão: Pacientes com RSCcPN tiveram chance de nova cirurgia três vezes maior do que aqueles sem pólipos nasais, no período seguido. Entre os pacientes com RSCsPN, apenas a asma foi um fator de pior prognóstico significativo, levando à chance de cirurgia 5,5 vezes maior do que os não-asmáticos. Já entre os pacientes com RSCcPN, aqueles com recidiva apresentaram maior extensão da doença à TC antes da primeira cirurgia. Foram ainda considerados fatores significativamente de pior prognóstico nos pacientes com RSCcPN: asma (odds ratio [OR] de 3,2); atopia a fungos (OR de 1,9); eosinofilia periférica (considerada como >500/µL, levando a OR de 1,9); e intolerância ao Ácido Acetil Salicílico (AAS) (DREA, apresentando OR de 2,5). Conclusões: Concluiu-se que a presença de pólipos per se é fator de pior prognóstico, aumentando em três vezes a chance de recorrência cirúrgica. Entre os pacientes com RSCsPN, apenas a asma influenciou o prognóstico. Já naqueles com RSCcPN, a asma, eosinofilia periférica, atopia a fungos e DREA aumentaram significativamente a probabilidade de nova intervenção cirúrgica. / Introduction: The analysis of prognostic factors associated with the recurrence of chronic rhinosinusitis (CRS), either with nasal polyps (CRSwNP) or without (CRSsNP), is still poorly discussed in the literature. Objective: To evaluate the patients that underwent endoscopic sinus surgery (ESS) due to CRS in Clinics Hospital of Ribeirão Preto Medical School, University of São Paulo, between 1996 and 2006, and to correlate the long-term recurrence to clinical factors, such as extensiveness of the disease, atopy, smoking habits, eosinophilia, and Aspirinexacerbated respiratory disease (AERD). Methods: We collected data of 201 patients, who were followed during an average period of 12 years. Clinical data collected were: extensiveness of the disease at endoscopy and at CT scans, prick test, blood exams, and pulmonary function. The follow-up period after surgery was assessed, and the indication of a new surgical procedure was considered as a poor prognostic factor. Comparison between factors was performed by Kaplan-Meyer curve, with Log-rank post-test. Results and discussion: CRSwNP patients were 3 times more likely to need a revisional surgery than CRSsNP during the follow-up period. Only asthma was a significant prognostic factor in patients with CRSsNP, leading to 5.5 times higher chance of recurrence than non-asthmatic patients. Among patients with CRSwNP, patients with recurrence presented, prior to surgery, higher CT scan extension of the disease. Other factors that influenced the prognosis on CRSwNP were: asthma (odds ratio [OR]: 3.2); atopy for fungi (OR: 1.9); peripheral eosinophilia (considered as >500/?L, leading to an OR: 1.9); and ASA intolerance (AERD; OR: 2.5). Conclusions: The presence of polyps were related to poor prognosis per se, leading to a higher chance of surgical recurrence. Among patients with CRSsNP, only asthma influenced the prognosis. Among the patients with CRSwNP, asthma, peripheral eosinophilia, fungi atopy, and AERD significantly increased the likelihood of further surgical intervention.