Browsing by Author "Vale-Pereira, S"
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- Apoptosis and asthma in the elderlyPublication . Todo-Bom, A; Mota-Pinto, A; Alves, V; Vale-Pereira, S; Santos-Rosa, MBACKGROUND: Asthma is a chronic inflammatory disorder of the airways. The persistence of airway inflammation depends on a decrease in apoptosis of T lymphocytes and eosinophils and survival of these activated cells. T lymphocytes expressing gamma delta receptors can be identified in human lungs and play an important role in immune defence against pathogens and in the regulation of chronic inflammation. Aging is associated with evidence of some immune dysregulation. OBJECTIVE: The aim of this study was to analyze the apoptosis receptors of T lymphocytes in long-lasting asthma, to establish their correlation with activation markers such as CD25+ and human leukocyte antigen (HLA)-DR+, and to analyze the gama delta T cell expression in this disease. METHODS: A group of 64 individuals (group A) who had had asthma for more than 30 years (mean age [+/-SD] 72 +/- 5 years) and 61 healthy individuals acting as controls--group B with 41 individuals (mean age 79 +/- 7 years) and group C with 20 individuals (mean age 38 +/- 12 years) were included in the study. All subjects underwent clinical evaluation and spirometric testing. Peripheral blood cells were stained with monoclonal antibodies anti-CD3, anti-CD4, anti-CD8, anti-CD25, anti-TCR gamma delta, anti-HLA-DR and anti-CD95. Statistical comparisons were performed between the asthmatics and the elderly control group and between the elderly control group and the adult control group. RESULTS: The average percentage of predicted forced expiratory volume in the first second was 73.6 gamma delta 25.3. The mean values of T cell receptors for asthma group A vs elderly control group B vs adult control group C respectively, were the following: CD3, 74.9+/-7 vs. 74.8 +/- 8.8 (P=ns) vs. 76.7 +/- 4.2 (P=ns); CD4, 48.8 +/- 8.7 vs. 43.5 +/- 10.2 (P=ns) vs. 44.8 +/- 3.8 (P=ns); CD8, 23.3 +/- 7.9 vs. 25.7 +/- 10.2 (P=ns) vs. 25.6 +/- 4.5 (P=ns); CD25, 14.3 +/- 5.9 vs. 22.4 +/- 7.8 (P = .0001) vs. 5.5 +/- 2.4 (P = .0001); TCR gamma delta, 2.8 +/- 2.1 vs. 4.1 +/- 3.3 (P < .05) vs. 4.6 +/- 2.1 (P=ns); HLA-DR, 18.4 +/- 9.2 vs. 17.8 +/- 5.9 (P=ns) vs. 15.4 +/- 5.1 (P=ns) and CD95, 49.3 +/- 13.7 vs. 52.6 +/- 12.1 (P=ns) vs. 13.8 +/- 10.8 (P = .0001). CONCLUSIONS: The immunological and inflammatory changes related to ageing may cause an increase in CD95 and CD25 T cell expression. In asthma, blood cells may express increased activation and apoptosis markers but in elderly patients taking steroids, these receptors remain within normal ranges. The number of gamma delta T cells may be lower in long-lasting asthma, and have a limited modulatory effect on allergic inflammatory reactions. The evaluation of patients with long-lasting asthma should take into account the immunological and inflammatory changes present in the elderly in order to avoid results being misinterpreted
- Asma: Reflexo da apoptose e de fenótipos de regulaçãoPublication . Todo-Bom, A; Mota-Pinto, A; Alves, V; Vale-Pereira, S; Chieira, S; Santos-Rosa, MA asma é uma doença inflamatória crónica das vias aéreas, e os linfócitos T desempenham um papel central na sua patogénese. Na inflamação alérgica quer os eosinófilos quer os linfócitos T podem ter um tempo de vida aumentado no local da inflamação, por redução da apoptose, mantendo-se activados. No pulmão encontra-se uma subclasse de linfócitos T que expressa receptor γδ. Estes linfócitos T γδ (TCRγδ) representam 1 a 10% dos linfócitos maduros circulantes e desempenham funções importantes na primeira linha de defesa imunitária contra agentes microbianos, na inflamação alérgica e na regulação da inflamação crónica. O objectivo deste estudo consistiu na análise de linfócitos T na asma persistente de evolução arrastada em doentes idosos no que concerne à expressão de receptores de apoptose, na sua correlação com marcadores de activação (CD25 e HLADR) e ainda à expressão TCR γδ nesta doença crónica comparativamente a indivíduos saudáveis do mesmo grupo etário e a indivíduos saudáveis adultos. Foram incluídos no estudo 64 doentes asmáticos (72±5 anos), com mais de 30 anos de evolução da doença e um grupo-controlo com 61 indivíduos, dividido em dois subgrupos de 41 indivíduos idosos (79±7 anos) e de 20 indivíduos adultos (38±12 anos). Todos os doentes foram avaliados do ponto de vista clínico e realizaram uma espirometria com Vitalograph Compact. As células do sangue periférico foram marcadas com anticorpos monoclonais anti-CD3, anti-CD4, anti-CD8, anti-CD25 anti-TCR γδ, anti-HLA-DR e anti- -CD95. A análise estatística direccionou-se a comparar asmáticos com controlo-idosos e controlo-idosos com controlo- -adultos. A percentagem média de forced expiratory volume in the first second (FEV1) foi de 73,6±25,3. Relativamente aos receptores estudados, obtiveram-se os seguintes resultados em asmáticos vs controlo-idosos vs controlo-adultos: CD3: 74,9±7 vs 74,8±8,8 vs 76,7±4,2; CD4: 48,8±8,7 vs 43,5±10,2 vs 44,8±3,8; CD8: 23,3±7,9 vs 25,7± 10,2 vs 25,6±4,5; CD25: 14,3±5,9 vs 22,4±7,8 (p=0,000) vs 5,5 ±2,4 (p=0,000); TCR γδ: 2,8±2,1 vs 4,1 ± 3,3 (p<0,05) vs 4,6±2,1; HLA-DR: 18,4± 9,2 vs 17,8± 5,9 vs 15,4± 5,1 e CD95: 49,3±13,7 vs 52,6±12,1 vs 13,8± 10,8 (p=0,000). Em conclusão, as alterações imunológicas e inflamatórias associadas à idade determinam um aumento da expressão nos linfócitos T de CD25 e de CD95. Na asma, a probabilidade de células do sangue periférico expressarem um maior número de marcadores de activação e uma redução de marcadores de apoptose pode ser limitada nas formas estáveis da doença, não introduzindo modificações significativas às determinadas pelo envelhecimento. Os linfócitos T γδ podem estar reduzidos na asma com evolução arrastada, podendo por isso exercer uma acção modesta na modulação da inflamação e contribuir para a cronicidade destas situações clínicas. Sendo a asma uma doença crónica irreversível qualquer análise ou interpretação fisiopatológica efectuada em formas tardias e arrastadas tem necessariamente de considerar as alterações decorrentes do processo de envelhecimento. A ausência de análise destes dados pode promover interpretações menos rigorosas, com ilações terapêuticas desajustadas.
- Determinação da neopterina e de defesas antioxidantes na asma de evolução arrastadaPublication . Mota-Pinto, A; Todo-Bom, A; Vale-Pereira, S; Alves, V; Santos-Rosa, MAsthma is a condition characterised by a chronic immunoinflammatory response to different triggers. Neopterin (NPT) is synthesised by human macrophages upon stimulation with interferon-gamma and is also capable of enhancing the oxidative potential of reactive oxygen species. NPT is useful for the monitoring of cell-mediated (Th1-type) immune activation. This study analysed the behaviour of NPT in long lasting asthma, considering its role as a marker of Th1 environment. Allergic parameters (skin prick tests, Immunoglobin E (IgE), and eosinophil count) and NPT were evaluated in an asthmatic group and in a control group. We also analysed the C Reactive Protein (CRP) concentration, Total Antioxidant Status (TAS) and Superoxide Dismutase Enzyme (SOD) in both groups. A group of individuals aged over 65 years old was selected. It included 64 asthmatic patients (72+/-5 years) and 41 healthy individuals (79+/-7 years). Blood cell counts showed statistically different median values of eosinophils (5.42+/-4.7 vs 2.8+/-2.8;p<.04), IgE (493.2+/-549.8 vs 85.3+/-194.UI/ml; p=.000) and NPT was non-statistically decreased (2.4+/-2.8 vs 4.0+/-4.7 ng/ml) in allergic asthmatic patients when compared with non-allergic asthmatic patients. Both allergic and non-allergic asthmatic patients presented a statistically significant decreased expression of TAS (0.84+/-0.14/0.86+/-0.11 vs 0.91+/-0.10 mM) and SOD (584.8+/-108.7/595.6+/-235.9 vs 822.9+/-179.5) when compared with normal control subjects. Our results suggest macrophage involvement in asthma pathogenesis. The deficit in antioxidant defence impacts negatively on this disease. The increase of NPT values in non-allergic asthma consolidates these affirmations and mapping this parameter should be part of the work of an analytical study panel as it may lead to allergic asthma being distinguished from non- allergic asthma.
- Doseamento das granzimas A e B na sarcoidose pulmonar (estudo experimental)Publication . Dourado, M; Bento, J; Mesquita, L; Marques, A; Vale-Pereira, S; Ribeiro, AB; Mota-Pinto, ASarcoidosis is a systemic disease of unknown aetiology, morphologically characterized by well-formed epithelioid granulomas, which show little or no central necrosis. These may be present in any organ or tissue. The lung is the most frequently and prominently involved target. The granuloma is often very sharply demarcated from the adjacent tissue and is surrounded by a mantle of lymphocytes, which mediate lysis of target cells by various mechanisms, including exocytosis of lytic proteins, perforins and granzymes. Sarcoidosis laboratorial diagnosis is usually made by SACE and Lisozyme dosages. The granzymes A and B could be two other markers of the disease, since the sarcoidosis granuloma is rich in cytotoxic and NK cells. An ELISA Kit was used to measure Granzyme A and B in serum of a normal control group (NC) (n=30), and in two groups with lung pathology: one without sarcoidosis, disease control (DC) (n=21) and other with sarcoidosis (S) (n=11). Our results showed that SACE activity is significantly augmented in S group comparing with NC and DC, respectively: 82,6+/-32,7/31,9+/-17,8 - p=0,00017 and 82,6+/-32,7/31,9+/-17,8 - p=0,00024. Lisozyme activity is significantly augmented in S and DC groups comparing with NC. Granzyme B showed a significant decrease in DC and S groups comparing with NC. Granzyme A showed a significant decrease between S/NC groups. Our results suggest that the decrease of Granzyme A and B in sarcoidotic patients could be related to an ineffective inflammatory local response related to the formation of sarcoidosis granulomas. More studies are needed, particularly in BAL.
- Elevated neopterin levels in non-allergic asthmaPublication . Mota-Pinto, A; Todo-Bom, A; Vale-Pereira, S; Alves, V; Santos-Rosa, MNeopterin is synthesized by human monocyte-derived macrophages upon stimulation with interferon-gamma (IFN-gamma). Measurement of neopterin concentration is useful to monitor cell-mediated (Th1-type) immune activation. In this study, we aimed to analyze the behaviour of neopterin in long lasting asthma considering its role as a marker of the Th1 environment and to establish the distinction between patients belonging either to the allergic or the non-allergic population, particularly in the elderly where asthma is often under diagnosed. Therefore we evaluated allergic parameters such as skin prick tests, IgE and hemogram (eosinophils count), and we compared our findings with neopterin values found in an age-matched control population. A group of individuals older than 65 was selected. It included 64 asthmatic patients (mean age 72+/-5 years) and 41 healthy individuals (mean age 79+/-7 years). In our study population, 42 patients presented positive skin tests, mainly to house dust mites. All patients were clinically stable and presented an average percentage of predicted forced expiratory volume in the first second (FEV1) of 73.6+/-25.3 and predicted median expiratory flow percentage (MEF50) of 38.8+/-26.7. Blood cell counts showed statistically different mean values of eosinophils between allergic and non-allergic controls (5.42+/-4.7% versus 2.8+/-2.8%; p<0.04). IgE values were increased in allergic asthmatic patients when compared with non-allergic asthmatic patients (493.2+/-549.8IU/ml versus 85.3+/-194.4IU/ml; p=0.000). Allergic asthmatic patients presented mean neopterin levels similar to those found in the control group (2.4+/-2.8ng/ml versus 2.1+/-1.9ng/ml). In contrast, in non-allergic asthmatic patients these values were higher when compared with the control group (4.0+/-4.7ng/ml versus 2.1+/-1.9ng/ml). Neopterin levels were lower in allergic asthmatic patients when compared with non-allergic asthmatic patients (2.4+/-2.8ng/ml versus 4.0+/-4.7ng/ml). Within asthmatic patients, those with higher neopterin values (>2.1ng/ml) presented lower mean IgE values (IgE
- FoxP3, GATA-3 and T-bet expression in elderly asthmaPublication . Vale-Pereira, S; Todo-Bom, A; Geraldes, L; Schmidt-Weber, C; Akdis, CA; Mota-Pinto, ABACKGROUND: Asthma is a chronic inflammatory disorder in which Th2, Th1 and suppressive T cells (Tregs) play a role. The transcription factor FoxP3 plays a role in Treg differentiation while T-bet is important for Th1 and GATA-3 for Th2 differentiation from naïve T cells. Recent data show that age-related deregulation of Treg cells is a mechanism of senescence affecting several chronic diseases. It is crucial to understand the behaviour of these cell populations in asthma for elderly patients. OBJECTIVE: To evaluate FoxP3, GATA-3 and T-bet gene expression under basal conditions and after in vitro activation in a group of elderly asthmatic compared with age-matched healthy individuals. METHODS: Thirty-two elderly asthmatics and 17 healthy elderly individuals were selected. Serum total IgE was measured, and peripheral blood mononuclear cells (PBMCs) were isolated and stimulated in vitro with anti-CD3/anti-CD28, followed by mRNA isolation. After reverse transcription, real-time quantitative PCR was performed and relative quantification was determined 2(-ΔΔCt)(2(-ΔΔCt) method). RESULTS: The mean values and standard deviation of FoxP3, GATA-3 and T-bet relative expression for control vs. asthma were 10.2±6.8 vs. 4.8±3.8, 2.4±2.9 vs. 1.7±0.9 and 3.3±2.1 vs. 2.1±1.5, respectively. Healthy individuals showed significantly higher expression of FoxP3 and T-bet; asthmatics had a lower T-bet/GATA-3 ratio, higher serum IgE and a positive significant correlation between total IgE and GATA-3 expression. CONCLUSION AND CLINICAL RELEVANCE: Elderly asthmatic patients have lower FoxP3 mRNA expression in PBMC, which can be associated with the sustained inflammatory process and with the decreased immune tolerance by Treg cells. The T-bet deficiency and the correlation of GATA-3 expression with the increase of IgE are characteristics of long-lasting asthma. Changes related to the immunosenescence process could provide an explanation for the minor differences observed between the groups. It is important to clarify persistent modifications in long-lasting asthma in the elderly and adequate future therapeutic approaches.
- Substance P in Long-Lasting Asthma: Immunoinflammatory pathwaysPublication . Todo-Bom, A; Mota-Pinto, A; Vale-Pereira, S; Alves, V; Dourado, M; Santos-Rosa, MBackground: Substance P (SP) was described at the beginning of the 20th Century, and its biological action was recognized to have implications in neurogenic inflammation and constriction of smooth muscles. The changes associated with inflammatory chronicity can compromise organ function reversibility. The role of neuromechanisms in the pathology of the disease has been investigated in order to achieve better diagnosis and therapeutic approaches. The stimulation of human cells, such as macrophages and polymorphonuclear cells by SP leads to their activation and to the release of reactive oxygen species (ROS) by these cells. Consequently, a continuous inflammatory disability is observed, mainly if a decrease in antioxidant defence occurs. SP is a substrate for dipeptidyl peptidase IV (DPPIV), which is a multifunctional molecule with enzymatic and proinflammatory activities. CD26 is considered an activation T cell marker. The aim of the present study was to analyse if serum SP values in long-lasting asthma patients were related to lung function parameters. It was also decided to analyze the relationship of SP with superoxide dismutase (SOD) and total antioxidant activity in serum (TAS), as well as its association to CD26/DPPIV values considering their immunological and inflammatory properties. Methods/Data base: A group of individuals older than 65 years, including 64 asthmatic patients (mean age 72±5 years) and 41 healthy individuals (mean age 79±7years) was selected. Both subgroups were submitted to clinical observation, to skin prick tests (SPT) and to SP, TAS, SOD, and DPPIV determinations. T cell CD26 typing was also performed. Lung function tests were done on all patients. Results: Among the patients studied, 42 had positive skin tests, mainly to house dust mites. Asthmatic patients showed a significant increase in SP values (116.2±138.9 vs 39.5±17.9 pg/ml) when compared to controls and a significant decrease in TAS levels (.85±.13 vs .91±.10 mM) and in SOD levels (588.1±156.l vs 822.9±179.5 U/gHb). All patients were clinically stable and presented an average percentage of predict forced expiratory volume in the first second (FEV1) of 73.6±25.3 and median expiratory flow percentage of predict (MEF50) of 38.8±26.7. DPPIV values were significantly increased in asthmatics compared to controls (69.7±15.2 vs 58.6±14.3 U/L). The CD26 expression was only slightly increased in asthmatic patients (41.9±10.2 vs 39.4±11.4). Conclusion: These results confirm the role of SP in asthma and give a contribution for a better knowledge of the immunoinflammatory pathway associated with this chronic disorder. A final goal for these studies would be to achieve a better therapeutic approach in order to improve the outcome of asthmatic patients.
- Substância P na asma brônquica: Intervenção nos mecanismos de oxidação e activação celularPublication . Todo-Bom, A; Mota-Pinto, A; Vale-Pereira, S; Alves, V; Dourado, M; Chieira, C; Santos-Rosa, MThe substance P (SP) was described in the beginning of the XXth Century, and its biological actions was recognized to have implications in neurogenic inflammation and constriction of smooth muscles. The bronchial asthma prevalence has been increasing in all developed countries. The changes associated to inflammatory chronicity can compromise the organ function reversibility. The role of neuromechanisms in the pathology of the disease has been investigated considering the hypotheses that better therapeutic approaches can be achieved. The stimulation of human cells by SP leads to their activation and to reactive oxygen species (ROS) release. Consequently a persisting inflammatory disability is observed, mainly if a decrease in the anti-oxidant defence occurs. SP is a substrate for dipeptidyl peptidase IV (DPPIV), which is a multifunctional molecule with enzymatic and proinflammatory activities. CD26 was identified as the membrane DPPIV and is considered as an activation T cell marker, as well as CD25. The aim of the present study was therefore to analyse if serum SP values in long-term asthma patients, were associated to lung function parameters, considering the role of SP in neurogenic inflammation and in bronchoconstriction. It was also proposed to analyse the relationship of SP with superoxide dismutase (SOD) and total anti-oxidation activity in serum (TAS), considering the ROS production during the inflammatory process, as well as its association to CD26 and CD25 values and their immunologic and inflammatory properties. A group of individuals older than 65 years including 64 asthmatic patients (mean age 72.4±5.1 years) and 41 healthy individuals (mean age 79.2±7.0 years) was selected. Both subgroups were submitted to clinical observation, to skin prick tests and to SP, TAS and SOD. T cell CD26 and CD25 typing was also performed. Lung function tests were done to all patients. Among patients studied, 42 presented positive skin tests, mainly to house dust mites. Asthmatic patients presented significant increased values of SP (116.2±138.9 vs 39.5±17.9 pg/ml) when compared to controls and a significant decrease of TAS (.85±.13 vs .91±.10 mM) and SOD (588.1±156.l vs 822.9±179.5 U/gHb). All patients have clinical stability and presented forced expiratory volume in one second (FEV1) values of 73.6±25.3 l/s and peak expiratory flow (PEF50) of 38.8±26.7l. The CD25 expression was significantly increased in disease (14.3±5.9 vs 22.4±7.8) while CD26 was only slightly increased (41.9±10.2 vs 39.4±11.4). These results confirm the role of SP in the respiratory pathology studied and give a contribution for a better knowledge of the network of immunoinflammatory pathway, associated to this chronic disorder. A final goal for these studies would be a better diagnostic and therapeutic approach in this pathology.