RESUMO
OBJECTIVE: Aspiration of duodenogastric refluxate may damage the respiratory epithelium of lung allografts in transplant recipients. We sought to define a mechanism by which aspiration of duodenogastric fluid augments the risk of bronchiolitis obliterans syndrome after lung transplant in a murine model. METHODS: We analyzed the immunological effects of acute aspiration of duodenogastric fluid (0.5 mL/kg) on transplant naive (strain DBA/2J) and transplanted mice (strain B6D2F1/J to strain DBA/2J). Serum antibodies to the lung self-antigens (SAgs) K-alpha1 tubulin and collagen-V were determined by enzyme-linked immunosorbent assay. Exosomes were isolated from serum, and immunoblot membranes were probed for antibodies to lung SAgs. Lung sections were assessed for fibrotic burden and obliterative bronchiolitis lesions by histologic and immunohistochemical analyses, including trichrome staining. RESULTS: Transplanted mice that received duodenogastric fluid developed higher levels of antibodies to the lung SAgs K-alpha1 tubulin and collagen-V and exosomes with lung SAgs on posttransplant days 14 and 28 than transplanted mice with sham aspiration or transplant naive mice (with and without aspiration). All lung allografts demonstrated severe grade A4 rejection on posttransplant day 14, with the highest mean fibrotic burden and mean number of obliterative bronchiolitis-like lesions per microscopic field on day 28 in recipients with aspiration. CONCLUSIONS: This study links aspiration of duodenogastric fluid after lung transplant to higher autoimmune responses to lung SAgs and the release of circulating exosomes with lung SAgs, which together promote sustained immune responses leading to extensive lung parenchymal damage and, ultimately, severe obliterative bronchiolitis-the histologic hallmark of bronchiolitis obliterans syndrome.
Assuntos
Síndrome de Bronquiolite Obliterante , Colágeno Tipo V , Transplante de Pulmão , Aspiração Respiratória de Conteúdos Gástricos , Tubulina (Proteína) , Animais , Camundongos , Autoantígenos/imunologia , Síndrome de Bronquiolite Obliterante/etiologia , Síndrome de Bronquiolite Obliterante/imunologia , Síndrome de Bronquiolite Obliterante/patologia , Colágeno Tipo V/imunologia , Suco Gástrico/imunologia , Rejeição de Enxerto , Secreções Intestinais/imunologia , Pulmão/imunologia , Pulmão/patologia , Transplante de Pulmão/efeitos adversos , Camundongos Endogâmicos DBA , Tubulina (Proteína)/imunologia , Aspiração Respiratória de Conteúdos Gástricos/complicações , Aspiração Respiratória de Conteúdos Gástricos/imunologiaRESUMO
BACKGROUND: Systemic sclerosis (SSc) is a connective tissue disorder which key feature is a fibrotic process. The role of Endothelin-1 (ET-1) and T-helper (Th)-1 cells in lung and skin fibrosis is well known, although Th17- and Treg-cells were found to be involved. However, no studies analyzed cytokines expression in gastric-juice of SSc patients. Our study aimed to evaluate proinflammatory and profibrotic cytokines in gastric-juice of SSc patients and to investigate their correlations with esophageal dysmotility. METHODS: Patients performed upper-gastrointestinal-endoscopy with gastric-juice collection, esophageal manometry and thoracic CT-scan. GM-CSF, ET-1, Th-1 (IFN-γ, IL-1ß, TNF-α, IL-2, IL-6, IL-9), Th-17 (IL-17, IL-21, IL-22, IL-23) and T-reg (IL-10, TGF-ß) related cytokines were measured in 29 SSc-patients and 20 healthy-controls. RESULTS: Patients showed significant lower levels of IL-6, IL-17, IL-22 and ET-1 (p < 0.005) compared with controls. Patients with atrophic gastritis presented significant lower levels of IL-2, IL-9, IL-6, TGF-ß, GM-CSF, IL-17 and ET-1 (p < 0.005) compared to patients without gastritis. Increased values of IL-2, IL-9, IL-1ß, IL-17, ET-1 and GM-CSF (p < 0.005) were observed in patients with esophageal impairment. This is the first report of cytokines measurement in gastric juice of patients with SSc. The high IL-17 concentrations in gastric-juice of scleroderma patients with esophageal dysmotility support the signature of Th-17 cells in scleroderma esophageal fibrosis.
Assuntos
Esôfago/imunologia , Suco Gástrico/imunologia , Interleucina-17/genética , Escleroderma Sistêmico/genética , Linfócitos T Reguladores/imunologia , Células Th1/imunologia , Células Th17/imunologia , Adulto , Estudos de Casos e Controles , Endotelina-1/genética , Endotelina-1/imunologia , Esôfago/patologia , Feminino , Suco Gástrico/química , Expressão Gênica , Humanos , Interleucina-17/imunologia , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-2/genética , Interleucina-2/imunologia , Interleucina-23/genética , Interleucina-23/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Interleucina-9/genética , Interleucina-9/imunologia , Interleucinas/genética , Interleucinas/imunologia , Pulmão/imunologia , Pulmão/patologia , Masculino , Pessoa de Meia-Idade , Escleroderma Sistêmico/imunologia , Escleroderma Sistêmico/patologia , Pele/imunologia , Pele/patologia , Estômago/imunologia , Estômago/patologia , Linfócitos T Reguladores/patologia , Células Th1/patologia , Células Th17/patologia , Tomografia Computadorizada por Raios X , Interleucina 22RESUMO
Helicobacter suis has a greater tendency to induce gastric mucosa-associated lymphoid tissue lymphoma compared with other Helicobacter species in humans and animals. Saccharomyces boulardii has been established as an adjunct to H. pylori eradication treatment, but the effect of S. boulardii administration alone on Helicobacter infection remains unclear. Here, we found that S. boulardii administration effectively decreased the bacterial load of H. suis and inhibited the formation of lymphoid follicles in the stomach post-infection. The levels of H. suis-specific immunoglobulin A (IgA) and secretory IgA in the gastric juice and small intestinal secretions and the production of mouse ß-defensin-3 in the small intestinal secretions were significantly increased by S. boulardii administration at 12 weeks after H. suis infection. In addition, feeding with S. boulardii inhibited the expression of inflammatory cytokines and lymphoid follicle formation-related factors after H. suis infection. These results suggested that S. boulardii may be useful for the prevention and treatment of Helicobacter infection-related diseases in humans.
Assuntos
Infecções por Helicobacter/microbiologia , Helicobacter heilmannii/fisiologia , Interações Microbianas/imunologia , Probióticos/administração & dosagem , Saccharomyces boulardii/fisiologia , Estômago/imunologia , Estômago/microbiologia , Animais , Citocinas/genética , Citocinas/metabolismo , Feminino , Suco Gástrico/imunologia , Imunoglobulina A/imunologia , Imunoglobulina A Secretora/imunologia , Mediadores da Inflamação/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/imunologia , Intestinos/microbiologia , Camundongos , Estômago/patologia , Fatores de Tempo , beta-Defensinas/biossínteseRESUMO
Gastroesophageal reflux disease (GERD) is defined with clinical symptoms of heart burning and regurgitation. It may be associated with external esophageal symptoms such as chronic cough, asthma, laryngitis, chronic lung disease, sinusitis and pulmonary fibrosis. In the present study, rats with chronic aspiration of gastroduodenal contents were studied for cellular phenotypes and cytokine concentrations in bronchoalveolar lavage and lung tissue. Thirty-six male Albino N-MRI rats were randomly divided into six groups. After anesthesia and tracheal intubation, the animals received either 0.5ml/kg of normal saline (control), gastric juice, pepsin, hydrochloric acid or bile salts by injection into their lungs twice a week for 8 weeks. In sham group nothing was injected. Thereafter, cellular phenotypes and cytokine concentrations of Interleukine (IL)-1α, IL-1ß, Transforming Growth Factor (TGF)-ß, Tumor Necrosis Factor (TNF)-α, and IL-6 were assessed in bronchoalveolar lavage and lung tissue homogenates. The numbers of epithelial cells, macrophages, neutrophils and lymphocytes in BAL and levels of cytokines IL-1α, IL-6, TNF-α and TGF-ß in BAL and lung tissue of test groups were significantly higher than the control group. Aspiration of bile salts caused more cytokine levels and inflammatory cells compared to other reflux components. It can be concluded that GERD with increased cytokines and inflammatory cells in lung could cause or exacerbate asthma and pulmonary fibrosis.
Assuntos
Ácidos e Sais Biliares/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/imunologia , Suco Gástrico/imunologia , Refluxo Gastroesofágico/imunologia , Animais , Asma/imunologia , Asma/patologia , Doença Crônica , Duodeno , Refluxo Gastroesofágico/patologia , Masculino , Fibrose Pulmonar/imunologia , Fibrose Pulmonar/patologia , Ratos , EstômagoRESUMO
UNLABELLED: To determine whether the levels of inflammatory mediators in gastric fluid (GF) of a premature newborn are associated with those in amniotic fluid (AF) of the newborn's mother. PATIENTS: Twenty-three pairs of pregnant women and their premature newborns <35 weeks gestation, born by Cesarean sections. METHODS: Amniotic fluids and newborn gastric fluids were obtained from women during Cesarean section procedure. The mother-premature newborn dyads were retrospectively assessed to analyze the clinical and laboratory data. Concentrations of interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-α) and mannose-binding lectin (MBL) were compared between amniotic and newborn gastric fluids in each dyad. RESULTS: Premature newborns and their mothers with funisitis had significantly higher median AF IL-6, TNF-α and GF IL-8 concentrations than those without funisitis (p = 0.022 for AF IL-6; p = 0.023 for AF TNF-α; p = 0.022 for GF IL-8). The concentrations of IL-6, IL-8, TNF-α and MBL in newborn GF were significantly correlated with those in AF in each dyad (p < 0.001, r = 0.872 for IL-6; p < 0.001, r = 0.851 for IL-8; p < 0.001, r = 0.768 for TNF-α; p < 0.001, r = 0.845 for MBL, respectively). CONCLUSION: The levels of inflammatory mediators in GF of a premature newborn immediately after birth are strongly associated with those in AF of the newborn's mother.
Assuntos
Líquido Amniótico/metabolismo , Suco Gástrico/metabolismo , Recém-Nascido Prematuro/metabolismo , Mediadores da Inflamação/metabolismo , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Adulto , Líquido Amniótico/imunologia , Corioamnionite/metabolismo , Feminino , Suco Gástrico/imunologia , Humanos , Recém-Nascido , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/imunologia , Estudos Retrospectivos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Stability in simulated gastric fluid is supposed to be an important parameter for the estimation of food allergenicity. In the present study, the digestive stability of allergenic protein tropomyosin (TM) and other food proteins from Grass prawn and Pacific white shrimp in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) digestion assay system was investigated and comparatively studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, and inhibition enzyme-linked immunosorbent assay (ELISA). RESULTS: In the SGF system, proteins such as actin and myosin heavy chain (MHC) were rapidly degraded within a short period of time, while TM was relatively resistant to pepsin digestion. In the SIF system, MHC was also easily decomposed, while TM and actin were resistant to digestion. Western blotting using a specific polyclonal antibody against TM indicated that the degradation pattern of shrimp TM by SGF and SIF was almost unaffected by the presence of other myofibrillar proteins. Further study by IgE immunoblotting and inhibition ELISA using sera from crustacean-allergic patients indicated that IgE binding of TM was decreased. CONCLUSION: Proteinase digestion is effective in reducing IgE binding of shrimp TM. It is also of interest to notice that Pacific white shrimp TM had higher digestion stability than Grass prawn TM. However, Pacific white shrimp TM revealed enhanced IgE binding over that of TM from Grass prawn and thus it is possibly more allergenic.
Assuntos
Digestão/imunologia , Hipersensibilidade Alimentar/metabolismo , Penaeidae , Alimentos Marinhos , Tropomiosina/metabolismo , Actinas/metabolismo , Adolescente , Animais , Western Blotting , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Hipersensibilidade Alimentar/imunologia , Suco Gástrico/imunologia , Suco Gástrico/metabolismo , Humanos , Imunoglobulina E/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/imunologia , Cadeias Pesadas de Miosina/metabolismo , Pepsina A/metabolismo , Tropomiosina/imunologia , Adulto JovemRESUMO
BACKGROUND: Patients with reflux-related respiratory symptoms are frequently treated with proton pump inhibitors (PPI). It is unclear whether aspiration of gastric juice (GJ) from patients "on" PPI can provoke a similar bronchial inflammatory reaction than that observed in patients "off" medication. The goal of this study was to evaluate the effect of GJ from patients with and without PPI treatment on production of IL-8 by human primary bronchial epithelial cells (PBEC). STUDY: PBEC were exposed during 24 hours to GJ (1/1000) from patients "on" (n=10) and "off" (n=13) PPI and to nonacidic gastric components (pepsin and bile acids). IL-8 concentration in supernatant was measured with enzyme-linked immunosorbent assay. Endotoxin level in GJ samples was analyzed with a LAL assay. RESULTS: Exposure of PBEC to GJ from patients "on" PPI provoked a higher production of IL-8 than GJ from patients "off" PPI [279 pg/mL (36 to 498) vs. 11 pg/mL (9 to 27)]. A correlation was found between pH of GJ and IL-8 production (r=0.659, P=0.0006). No correlation was found between IL-8 production and concentration of bile acids or pepsin. Filtration (0.20 [mu]m) of GJ from patients "on" PPI reduced IL-8 production. A positive correlation was found between IL-8 production and endotoxin levels of GJ samples (1/1000) (r=0.654, P=0.0007). CONCLUSIONS: Exposure of bronchial epithelial cells to GJ from patients "on" PPI is able to induce high IL-8 production. These results suggest that aspiration of GJ in patients treated with PPI might still be able to provoke a significant bronchial inflammatory reaction.
Assuntos
Brônquios/imunologia , Células Epiteliais/imunologia , Suco Gástrico/imunologia , Mucosa Gástrica/efeitos dos fármacos , Refluxo Gastroesofágico/tratamento farmacológico , Mediadores da Inflamação/metabolismo , Interleucina-8/metabolismo , Inibidores da Bomba de Prótons/uso terapêutico , Idoso , Ácidos e Sais Biliares/imunologia , Células Cultivadas , Endotoxinas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Ácido Gástrico/metabolismo , Suco Gástrico/metabolismo , Mucosa Gástrica/metabolismo , Refluxo Gastroesofágico/imunologia , Refluxo Gastroesofágico/metabolismo , Gastroscopia , Humanos , Concentração de Íons de Hidrogênio , Masculino , Pessoa de Meia-Idade , Pepsina A/imunologiaRESUMO
Leguminous crops are the main source of protein in Asian subcontinent including India and their proteins may induce allergic reactions in sensitized individuals. Pepsin resistance of proteins is a characteristic feature of most of the allergens. Simulated gastric fluid (SGF) assay as validated by digestion of purified known allergenic and non-allergenic proteins was the basis of this study. Purified allergenic proteins were stable to SGF digestion contrary to rapidly digested non-allergenic proteins. Crude proteins extracts (CPE) of soybean, peanut, chickpea, black gram, kidney bean and Bengal gram were digested in vitro to detect their non-digestible proteins. Six proteins from soybean and seven from peanut remained undigested after SGF digestion. Likewise, seven proteins from chickpea (70, 64, 55, 45, 35, 20 and 18 kDa), ten from black gram (47, 30, 29, 28, 26, 24, 22, 16, 14 and 12 kDa), five from kidney bean (45, 29, 24, 20 and 6.5 kDa) and one from Bengal gram (20 kDa) remained undigested in SGF. Most of the proteins stable in SGF for more than 2 min showed similarity with characterized allergens on the basis of their molecular weights as in case of soybean, peanut, chickpea and black gram. Also, soybean and chickpea stable proteins showed IgE binding property with respective allergic patient's sera. The non-digestible proteins from the chickpea, black gram, kidney bean and Bengal gram are being reported for the first time by our group. IgE binding of SGF resistant soybean and chickpea proteins is being reported first time as well.
Assuntos
Antígenos de Plantas/imunologia , Proteínas Alimentares/imunologia , Fabaceae/imunologia , Hipersensibilidade Alimentar/imunologia , Suco Gástrico/imunologia , Proteínas de Plantas/imunologia , Antígenos de Plantas/metabolismo , Proteínas Alimentares/metabolismo , Digestão/imunologia , Fabaceae/metabolismo , Suco Gástrico/metabolismo , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Proteínas de Plantas/metabolismoRESUMO
The objectives of this study were to formulate a vaccine based upon the different species/strains of methanogens present in sheep intended to be immunized and to determine if a targeted vaccine could be used to decrease the methane output of the sheep. Two 16S rRNA gene libraries were used to survey the methanogenic archaea in sheep prior to vaccination, and methanogens representing five phylotypes were found to account for >52% of the different species/strains of methanogens detected. A vaccine based on a mixture of these five methanogens was then formulated, and 32 sheep were vaccinated on days 0, 28, and 103 with either a control or the anti-methanogen vaccine. Enzyme-linked immunosorbent assay analysis revealed that each vaccination with the anti-methanogen formulation resulted in higher specific immunoglobulin G titers in plasma, saliva, and rumen fluid. Methane output levels corrected for dry-matter intake for the control and treatment groups were not significantly different, and real-time PCR data also indicated that methanogen numbers were not significantly different for the two groups after the second vaccination. However, clone library data indicated that methanogen diversity was significantly greater in sheep receiving the anti-methanogen vaccine and that the vaccine may have altered the composition of the methanogen population. A correlation between 16S rRNA gene sequence relatedness and cross-reactivity for the methanogens (R(2) = 0.90) also exists, which suggests that a highly specific vaccine can be made to target specific strains of methanogens and that a more broad-spectrum approach is needed for success in the rumen. Our data also suggest that methanogens take longer than 4 weeks to adapt to dietary changes and call into question the validity of experimental results based upon a 2- to 4-week acclimatization period normally observed for bacteria.
Assuntos
Archaea/crescimento & desenvolvimento , Archaea/imunologia , Biodiversidade , Metano/metabolismo , Rúmen/microbiologia , Ovinos/microbiologia , Vacinas/imunologia , Animais , Anticorpos/análise , DNA Arqueal/química , DNA Arqueal/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Suco Gástrico/imunologia , Imunoglobulina G/análise , Dados de Sequência Molecular , Plasma/imunologia , RNA Ribossômico 16S/química , RNA Ribossômico 16S/genética , Rúmen/imunologia , Saliva/imunologia , Análise de Sequência de DNA , Ovinos/imunologiaRESUMO
Stability in simulated gastric fluid has been suggested as a parameter for consideration in the allergenicity assessment of transgenic proteins. However, the relationship between the stability of proteins in simulated gastric fluid and allergenicity has been inconsistent among studies conducted with reference allergens and non-allergens. Differences in laboratory methods and data interpretation have been implicated as possible causes for conflicting study results. We attempted to mitigate some of the methodological inconsistencies among laboratory methods by applying a kinetic interpretation to results of digestion experiments conducted with a set of known allergens and putative non-allergens. We found that pepsinolysis in simulated gastric fluid generally followed an exponential (pseudo-first-order) pattern of decay, at least during the terminal (slower) phase of digestion, allowing the calculation of digestion half-lives. While digestibility estimates were reproducible and robust, results for the proteins evaluated in this study did not support a significant association between stability in simulated gastric fluid and allergenicity.
Assuntos
Alérgenos/metabolismo , Proteínas Alimentares/metabolismo , Suco Gástrico/imunologia , Proteínas Alimentares/imunologia , Digestão/fisiologia , Eletroforese em Gel de Poliacrilamida/métodos , Suco Gástrico/metabolismo , Meia-Vida , HumanosRESUMO
BACKGROUND: It is a general belief that a food allergen should be stable to gastric digestion. Various acidic plant polysaccharides, including pectin, are ubiquitous in fruit matrixes and can form hydrogels under low-pH conditions. OBJECTIVE: The purpose of this study was to investigate the effect of hydrogel forming polysaccharide-rich fruit matrixes on in vivo gastric and in vitro pepsic digestion of fruit allergens. METHODS: Fruit extract proteins (kiwi, banana, apple and cherry) and a purified major kiwi allergen Act c 2 were digested with simulated gastric fluid in accordance with the US Pharmacopeia. In vivo experiments on kiwi fruit digestion were performed on four healthy non-atopic volunteers by examining the gastric content 1 h after ingestion of kiwi fruit. The Act c 2 and kiwi proteins were detected in immunoblots using monoclonal anti-Act c 2 antibodies and rabbit polyclonal antisera. RESULTS: Crude fruit extracts were resistant to digestion by pepsin when compared with commonly prepared extracts. In the gastric content of all volunteers, following kiwi fruit ingestion and immunoblotting, intact Act c 2 was detected with anti-Act c 2 monoclonal antibodies, while kiwi proteins of higher molecular weights were detected using rabbit polyclonal antisera. Addition of apple fruit pectin (1.5% and 3%) to the purified kiwi allergen was able to protect it from pepsin digestion in vitro. CONCLUSION: The matrix effect in pectin-rich fruits can influence the digestibility of food proteins and thereby the process of allergic sensitization in atopic individuals.
Assuntos
Alérgenos/metabolismo , Digestão/imunologia , Hipersensibilidade Alimentar/imunologia , Frutas/imunologia , Pepsina A/fisiologia , Actinidia/química , Actinidia/imunologia , Animais , Dissacarídeos/análise , Frutas/química , Suco Gástrico/imunologia , Humanos , Camundongos , Monossacarídeos/análise , Pectinas/análise , Pepsina A/antagonistas & inibidores , Extratos Vegetais/imunologia , Proteínas de Plantas/análise , Proteínas de Plantas/metabolismo , CoelhosRESUMO
BACKGROUND: Pathological alteration in gastric mucosa is caused by Helicobacter pylori infection and is detectable by histological analysis. In particular, the alteration of gland mucous cells (GMCs)-type mucin, which plays a protective role against H. pylori infection, is critical in the pathogenesis of H. pylori-related gastritis. We established an assay for GMCs-type mucin and quantitatively assessed the pathophysiological changes in its content in human gastric juice samples. METHODS: The assay method for GMCs-type mucin was based on ELISA using a monoclonal antibody (HIK1083), and was used it to measure GMCs-type mucin in gastric juice obtained from patients with or without H. pylori infection. RESULTS: All the basic characteristics of the current method were satisfactory to quantify the GMCs-type mucin content in gastric juice. The GMCs-type mucin content, but not total mucin content, was significantly higher in patients with H. pylori infection (n=17; 437+/-476 U, mean+/-SD) than in those without H. pylori infection (n=55; 168+/-322 U, p<0.05). CONCLUSIONS: The current method is suitable for the quantitative analysis of GMCs-type mucin in gastric juice. The change in GMCs-type mucin content in gastric juice may be possibly implicated in the pathophysiology of the gastric mucosa and in the patient's gastric mucosal lesions.
Assuntos
Anticorpos Monoclonais/imunologia , Suco Gástrico/metabolismo , Mucinas Gástricas/análise , Mucosa Gástrica/metabolismo , Mucosa Gástrica/fisiopatologia , Gastropatias/metabolismo , Gastropatias/fisiopatologia , Animais , Calibragem , Ensaio de Imunoadsorção Enzimática , Suco Gástrico/imunologia , Mucinas Gástricas/imunologia , Mucinas Gástricas/metabolismo , Mucosa Gástrica/imunologia , Humanos , Concentração de Íons de Hidrogênio , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Gastropatias/imunologia , Suínos , TemperaturaRESUMO
Colonization with Helicobacter pylori eventuates in varied clinical outcomes, which relate to both bacterial and host factors. Here we examine the relationships between cagA status, serum and gastric juice antibody responses, and gastric inflammation in dyspeptic patients. Serum, gastric juice, and gastric biopsy specimens were obtained from 89 patients undergoing endoscopy. H. pylori colonization and cagA status were determined by histology, culture, and PCR methods, and acute inflammation and chronic inflammation in the gastric mucosa were scored by a single pathologist. Serum and gastric juice antibodies to H. pylori whole-cell and CagA antigens were determined by enzyme-linked immunosorbent assay. Relationships between variables were sequentially analyzed using univariate and multivariate statistical methods. Of the 89 subjects, 62 were colonized by H. pylori. By univariate analyses, levels of serum immunoglobulin G (IgG) and IgA and gastric juice IgA antibodies against whole-cell and CagA antigens each were significantly higher in the H. pylori-positive group than in the H. pylori-negative group (P<0.001). H. pylori and CagA sero-positivities were both significantly associated with enhanced inflammation in gastric antrum and body (P<0.02). The presence of gastric juice antibodies to H. pylori antigens was associated with more severe gastric inflammation. However, in multivariate analyses, only the presence of serum antibodies against CagA and, to a lesser extent, whole-cell antigens remained significantly associated with acute and chronic inflammation in antrum and body (P<0.05). Thus, serum antibody response to CagA correlates with severity of gastric inflammation. Furthermore, given the relationships demonstrated by multivariate analysis, determination of gastric juice antibodies may provide a better representation of serum, rather than secretory, immune response.
Assuntos
Anticorpos Antibacterianos/análise , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Suco Gástrico/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Adulto , Análise de Variância , Anticorpos Antibacterianos/sangue , Feminino , Suco Gástrico/microbiologia , Gastrite/classificação , Gastrite/imunologia , Gastrite/microbiologia , Infecções por Helicobacter/microbiologia , Humanos , Imunoglobulina G/análise , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da PolimeraseRESUMO
AIM: To testify the immunogenicity of a conservative B-cell linear epitope of Helicobacter pylori (H pylori) flagellin A. METHODS: Different programs were used to analyze the secondary structure, molecular hydropathy, and surface accessibility of H pylori flagellin A. Linear B-cell epitopes were estimated based on the structural and physiochemical information. Analysis of residue divergence was proposed to screen a conservative linear epitope. The 29-peptide (Pep29mer) synthesized by chemical method, including the predicted conservative B-cell epitope and a known K2d compatible T-cell epitope, was used to immunize mice, and then H pylori-specific antibodies were detected by ELISA. RESULTS: Based on the analyses of divergent amino acid residues, structural and physiochemical characteristics, it was strongly suggested that the short fragment NDSDGR was the core of a conservative linear epitope in flagellin A. Animals immunized by Pep29mer acquired efficient immune response. In detail, serum H pylori-specific IgA and IgG1 increased significantly in immunized group, while IgG2a only had an insignificant change. H pylori-specific IgA in gastrointestinal flushing fluid also increased significantly. CONCLUSION: The conservative short fragment NDSDGR is the core of a linear B-cell epitope of flagellin A.
Assuntos
Flagelina/imunologia , Helicobacter pylori/imunologia , Sequência de Aminoácidos , Animais , Antígenos de Bactérias/imunologia , Ensaio de Imunoadsorção Enzimática , Epitopos/imunologia , Suco Gástrico/imunologia , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Camundongos , Dados de Sequência MolecularRESUMO
This case control study presents data on the concentrations of nitrite and nitrate and a variety of pro-inflammatory cytokines such as interleukin-1 beta (IL-1 beta), interleukin-2R (IL-2R), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor TNF-alpha in gastric fluid and serum. Patients with gastritis, gastric ulcer and gastric cancer are studied and grouped according to infection by Helicobacter pylori. The 208 patients who underwent upper gastrointestinal endoscopic examination were classified as follows; H. pylori-positive gastritis (n = 32), H. pylori-negative gastritis (n = 32), H. pylori-positive ulcers (n = 34), H. pylori-negative ulcers (n = 34), 43 patients with H. pylori-positive gastric cancer in addition to 33 H. pylori-negative healthy control individuals. Gastric fluids and blood samples were taken concomitantly. Cytokines and nitrite and nitrate determinations were attempted as soon as possible after collection of the samples. Nitrite and nitrate levels of serum and gastric fluids of H. pylori-positive gastritis and ulcers were higher than H. pylori-negative gastritis and ulcers. The concentrations of total nitrite and nitrate and cytokines (TNF-alpha, IL-2R, IL-6, and IL-8) in gastric fluids and sera of H. pylori-positive gastric cancer patients were higher than H. pylori-negative control groups. IL-1 beta level was significantly elevated in gastric fluid of infected cancer patients but not in serum. Taken together, the results suggest that an increase in cytokine-NO combination in gastric mucosa previously reported by many studies is not restricted to local infected gastric tissue but also detected in gastric fluid and sera of H. pylori-positive subjects and may have an important role in the pathogenesis and development of common gastric diseases.
Assuntos
Citocinas/sangue , Suco Gástrico/metabolismo , Gastrite/metabolismo , Infecções por Helicobacter/metabolismo , Helicobacter pylori , Nitratos/sangue , Feminino , Suco Gástrico/imunologia , Mucosa Gástrica/imunologia , Mucosa Gástrica/metabolismo , Mucosa Gástrica/microbiologia , Gastrite/imunologia , Gastrite/microbiologia , Infecções por Helicobacter/complicações , Infecções por Helicobacter/imunologia , Humanos , Masculino , Óxido Nítrico/metabolismo , Neoplasias Gástricas/complicações , Neoplasias Gástricas/imunologia , Neoplasias Gástricas/metabolismo , Úlcera Gástrica/imunologia , Úlcera Gástrica/metabolismo , Úlcera Gástrica/microbiologiaRESUMO
BACKGROUND AND AIMS: The acid inhibitory effect of proton pump inhibitors is reported to be greater in the presence than in the absence of an H. pylori infection. This study was undertaken to test the hypothesis that the acid inhibitory effect of omeprazole given twice a day is greater in H. pylori infected healthy volunteers than in the same individuals following eradication because of differences in the pharmacodynamics of omeprazole, greater duodenogastric reflux, the effects of ammonia produced by the H. pylori, or lower gastric juice concentrations of selected cytokines, which may inhibit gastric acid secretion. MATERIALS AND METHODS: We undertook 24-hour pH-metry in 12 H. pylori-positive healthy volunteers: (1) when on no omeprazole; (2) when on omeprazole 20 mg bid for 8 days; (3) 2 months after eradication of H. pylori and when on no omeprazole; and (4) after eradication of H. pylori and when on omeprazole 20 mg twice a day. RESULTS: In subjects given omeprazole, eradication of H. pylori reduced pH and percentage pH >or= 3, as well as increasing the area under the H+ concentration-time curve. These differences were not due to alterations in (1) gastric juice concentrations of IL-1alpha, IL-8, IL-13, epidermal growth factor, or bile acids; (2) serum gastrin concentrations; or (3) the pharmacokinetics of omeprazole. There was no change in the difference in the H+ concentration-time curve 'without omeprazole' minus 'with omeprazole', when comparing 'after' versus 'before' eradication of H. pylori. CONCLUSIONS: Eradication of H. pylori was not associated with an alteration in the acid inhibitory potency when comparing the difference in gastric acidity 'with' versus 'without' omeprazole. When the results were expressed by simply taking into account the acid measurements while on omeprazole before versus after eradication of H. pylori, the acid inhibition with omeprazole was greater in the presence than in the absence of a H. pylori infection. The clinical significance of the small difference is not clear.
Assuntos
Antiácidos/administração & dosagem , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/metabolismo , Helicobacter pylori , Omeprazol/administração & dosagem , Inibidores da Bomba de Prótons , Adulto , Antiácidos/farmacocinética , Antibacterianos , Ácidos e Sais Biliares/metabolismo , Citocinas/metabolismo , Esquema de Medicação , Quimioterapia Combinada/uso terapêutico , Feminino , Suco Gástrico/imunologia , Suco Gástrico/metabolismo , Gastrinas/sangue , Infecções por Helicobacter/imunologia , Humanos , Concentração de Íons de Hidrogênio , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , Omeprazol/farmacocinéticaRESUMO
BACKGROUND: It has been reported that treatment with proton pump inhibitors (PPI) leads to partial elimination and suppression of Helicobacter pylori. In theory, since acid is known to denature immunoglobulins, this antibacterial activity of PPI may be due to a reduction in the acid output favouring humoral immunity. MATERIALS AND METHODS: We analysed prospectively fasting gastric juice in 54 consecutive patients attending upper endoscopy for pH and levels of IgG, IgA and IgM. In addition, two antral and two corpus biopsies were obtained and histologically examined for the presence of H. pylori. RESULTS: 41/54 patients were infected with H. pylori. Immunoglobulines in the gastric juice of these patients were found in 25/41 (IgG), 27/41 (IgA), and 29/41 (IgM) patients. There was a highly significant difference in the gastric pH when H. pylori infected patients with measurable IgG, IgA, or IgM were compared with those in whom no immunoglobulines were found (median pH: 6 vs. 2 in each group; p <.001) CONCLUSIONS: There is a close correlation between a high gastric pH and the presence of IgG, IgA, and IgM antibodies. Hence, it may be speculated that the efficacy of humoral immunity following H. pylori infection depends on a high pH such as resulting from PPI treatment.
Assuntos
Ácido Gástrico/metabolismo , Suco Gástrico/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Imunoglobulinas/análise , Adulto , Idoso , Feminino , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Inibidores da Bomba de PrótonsRESUMO
BACKGROUND: Gastrointestinal symptoms in cystic fibrosis are frequent, but little is known about the underlying pathophysiology. Mucosal secretion of IgA is important for the immunologic function in the human gastrointestinal tract but has not been studied in cystic fibrosis. The aim of this study was to quantify the release of IgA by the gastric mucosa in relation to interdigestive motor activity in patients with cystic fibrosis with different genotypes. METHODS: The study included 7 healthy adult volunteers and 10 adult patients with cystic fibrosis, all Helicobacter pylori-negative. All patients had pathological sweat tests and clinical symptoms and signs of cystic fibrosis. All but one were colonized with Pseudomonas aeruginosa. Three patients were pancreatic sufficient. The investigation was performed using intragastric perfusion and gastroduodenal manometry. RESULTS: During the investigation, 8 of 10 patients with cystic fibrosis showed the characteristic pattern of interdigestive motility. The patients had significantly lower levels of gastric IgA compared to healthy subjects during phases II and III of migrating motor complex, median (range) 120 (67-442) and 36 (6-299) microg/5 min. 382 (40-1176) and 56 (4-398) (P = 0.03 and P = 0.04), respectively. Only one patient with genotype R668C/unknown showed IgA levels within the normal range. There was no correlation to gastric presence of duodenogastric reflux markers. CONCLUSION: The interdigestive motility pattern was normal in most patients with cystic fibrosis. The low levels of IgA released from the gastric mucosa in the patients might indicate a defective gastric transmucosal IgA transport in cystic fibrosis.
Assuntos
Fibrose Cística/imunologia , Suco Gástrico/imunologia , Imunoglobulina A Secretora/metabolismo , Complexo Mioelétrico Migratório/imunologia , Adulto , Feminino , Motilidade Gastrointestinal/imunologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Helicobacter pylori, the main cause of gastritis and peptic ulcer, has been associated with idiopathic chronic urticaria (ICU), an immunological skin disorder of unknown origin. Eosinophilic cationic protein (ECP) is a cytotoxic molecule secreted by the activated eosinophils involved in the pathogenesis of ICU. We assessed serum/gastric juice ECP levels and gastric mucosal eosinophil infiltration in ICU patients infected or not with H. pylori and evaluated the modification after bacterium eradication. METHODS: 33 patients with ICU and 25 dyspeptic controls underwent upper gastrointestinal endoscopy for histological evaluation and assessment of H. pylori infection. One-week triple therapy was given to H. pylori-positive patients. Serum and gastric juice ECP levels, eosinophil infiltration from gastric mucosal sections and urticaria symptoms were evaluated in all patients at enrollment and 8 weeks after eradication. RESULTS: 19 of 33 (57%) ICU patients and 16 of 25 (64%) controls were found to be infected with H. pylori. Serum ECP was significantly higher in ICU patients compared to controls, regardless of infectious status. Gastric juice ECP and gastric eosinophil infiltration were significantly higher in infected ICU patients when compared both to uninfected ICU patients and controls. H. pylori eradication determined a significant decrease in gastric juice ECP and gastric eosinophil infiltration only in ICU patients. Moreover, a total or partial remission of urticaria symptoms was observed only in ICU patients in whom the bacterium was eradicated. CONCLUSIONS: Although H. pylori infection affects gastric juice ECP and eosinophil infiltration of ICU patients, the role of the bacterium in the pathogenesis of this skin disorder still remains uncertain.
Assuntos
Alérgenos/análise , Proteínas Sanguíneas/análise , Suco Gástrico/imunologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/imunologia , Ribonucleases , Urticária/microbiologia , Adulto , Antibacterianos/uso terapêutico , Doença Crônica , Proteínas Granulares de Eosinófilos , Eosinófilos/química , Eosinófilos/metabolismo , Feminino , Mucosa Gástrica/imunologia , Mucosa Gástrica/microbiologia , Infecções por Helicobacter/complicações , Humanos , Masculino , Urticária/sangueRESUMO
Episialin (MUC 1 antigen) is a membrane bound mucin. It is located on the surface of the epithelium. It can also be detected in soluble form in body fluids. In our paper we present method of the purification of this antigen from human gastric juice. MUC 1 antigen was isolated from human gastric juice by the gel filtration on Sepharose 4B column and affinity chromatography with HMFG-1 antibody coupled to CNBr activated Sepharose 4B. Using presented methods for purification of MUC 1 antigen from gastric juice we received after immunostaining one diffuse band of about 400 kDa molecular weight. Our results revealed that presented method of isolation of MUC 1 antigen from gastric juice is a valuable and sufficient procedure for purification of soluble form of this antigen.