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1.
Clin Exp Immunol ; 167(2): 356-65, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22236013

RESUMO

One of the promising approaches in the therapy of ulcerative colitis is administration of butyrate, an energy source for colonocytes, into the lumen of the colon. This study investigates the effect of butyrate producing bacterium Clostridium tyrobutyricum on dextran sodium sulphate (DSS)-induced colitis in mice. Immunocompetent BALB/c and immunodeficient severe combined immunodeficiency (SCID) mice reared in specific-pathogen-free (SPF) conditions were treated intrarectally with C. tyrobutyricum 1 week prior to the induction of DSS colitis and during oral DSS treatment. Administration of DSS without C. tyrobutyricum treatment led to an appearance of clinical symptoms - bleeding, rectal prolapses and colitis-induced increase in the antigen CD11b, a marker of infiltrating inflammatory cells in the lamina propria. The severity of colitis was similar in BALB/c and SCID mice as judged by the histological damage score and colon shortening after 7 days of DSS treatment. Both strains of mice also showed a similar reduction in tight junction (TJ) protein zonula occludens (ZO)-1 expression and of MUC-2 mucin depression. Highly elevated levels of cytokine tumour necrosis factor (TNF)-α in the colon of SCID mice and of interleukin (IL)-18 in BALB/c mice were observed. Intrarectal administration of C. tyrobutyricum prevented appearance of clinical symptoms of DSS-colitis, restored normal MUC-2 production, unaltered expression of TJ protein ZO-1 and decreased levels of TNF-α and IL-18 in the descending colon of SCID and BALB/c mice, respectively. Some of these features can be ascribed to the increased production of butyrate in the lumen of the colon and its role in protection of barrier functions and regulation of IL-18 expression.


Assuntos
Butiratos/metabolismo , Clostridium tyrobutyricum/fisiologia , Colite Ulcerativa/microbiologia , Interleucina-18/biossíntese , Probióticos/uso terapêutico , Fator de Necrose Tumoral alfa/biossíntese , Doença Aguda , Administração Retal , Animais , Translocação Bacteriana , Antígeno CD11b/biossíntese , Antígeno CD11b/genética , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/genética , Colite Ulcerativa/imunologia , Colite Ulcerativa/patologia , Colo/metabolismo , Colo/microbiologia , Colo/patologia , Sulfato de Dextrana/toxicidade , Ácidos Graxos/metabolismo , Imunocompetência , Interleucina-18/genética , Proteínas de Membrana/biossíntese , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Mucina-2/biossíntese , Mucina-2/genética , Mucinas/biossíntese , Fosfoproteínas/biossíntese , Fosfoproteínas/genética , Imunodeficiência Combinada Severa/genética , Imunodeficiência Combinada Severa/imunologia , Organismos Livres de Patógenos Específicos , Fator de Necrose Tumoral alfa/genética , Proteína da Zônula de Oclusão-1
2.
Physiol Res ; 71(3): 357-368, 2022 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-35616045

RESUMO

Growing evidence suggests that diabetes mellitus is associated with impairment of the intestinal barrier. However, it is not clear so far if the impairment of the intestinal barrier is a consequence of prolonged hyperglycemia or the consequence of external factors influencing the gut microbiota and intestinal mucosa integrity. Aim of the study was to perform an estimation of relationship between serological markers of impairment of the intestinal barrier: intestinal fatty acid-binding protein (I-FABP), cytokeratin 18 caspase-cleaved fragment (cCK-18), and soluble CD14 (sCD14) and markers of prolonged hyperglycemia, such as the duration of diabetes mellitus and glycated hemoglobin (HbA1c) via a correlation analysis in patients with diabetes mellitus. In 40 adult patients with type 1 diabetes mellitus and 30 adult patients with type 2 diabetes mellitus the estimation has been performed. Statistically significant positive correlation was found between cCK-18 and HbA1c (r=0.5047, p=0.0275) in patients with type 1 diabetes mellitus with fading insulitis (T1D). In patients with type 1 diabetes mellitus with ongoing insulitis (T1D/INS) and in patients with type 2 diabetes mellitus (T2D), no statistically significant positive correlations were found between serological markers of intestinal barrier impairment (I-FABP, cCK-18, sCD14) and duration of diabetes or levels of HbA1c. Similarly, in cumulative cohort of patients with T1D/INS and patients with T1D we revealed statistically positive correlation only between HbA1c and cCK-18 (r=0.3414, p=0.0311). Surprisingly, we found statistically significant negative correlation between the duration of diabetes mellitus and cCK-18 (r=-0.3050, p=0.0313) only in cumulative group of diabetic patients (T1D, T1D/INS, and T2D). Based on our results, we hypothesize that the actual condition of the intestinal barrier in diabetic patients is much more dependent on variable interactions between host genetic factors, gut microbiota, and environmental factors rather than effects of long-standing hyperglycemia (assessed by duration of diabetes mellitus or HbA1c).


Assuntos
Diabetes Mellitus Tipo 1 , Diabetes Mellitus Tipo 2 , Hiperglicemia , Adulto , Biomarcadores , Diabetes Mellitus Tipo 1/diagnóstico , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 2/diagnóstico , Hemoglobinas Glicadas/análise , Humanos , Receptores de Lipopolissacarídeos
3.
Clin Exp Immunol ; 163(2): 250-9, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21087444

RESUMO

Commensal bacteria have been shown to modulate the host mucosal immune system. Here, we report that oral treatment of BALB/c mice with components from the commensal, Parabacteroides distasonis, significantly reduces the severity of intestinal inflammation in murine models of acute and chronic colitis induced by dextran sulphate sodium (DSS). The membranous fraction of P. distasonis (mPd) prevented DSS-induced increases in several proinflammatory cytokines, increased mPd-specific serum antibodies and stabilized the intestinal microbial ecology. The anti-colitic effect of oral mPd was not observed in severe combined immunodeficient mice and probably involved induction of specific antibody responses and stabilization of the intestinal microbiota. Our results suggest that specific bacterial components derived from the commensal bacterium, P. distasonis, may be useful in the development of new therapeutic strategies for chronic inflammatory disorders such as inflammatory bowel disease.


Assuntos
Antígenos de Bactérias/administração & dosagem , Bacteroides/imunologia , Colite/terapia , Metagenoma/imunologia , Doença Aguda , Administração Oral , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Doença Crônica , Citocinas/sangue , Citocinas/imunologia , Feminino , Mucosa Intestinal/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID
4.
Allergy ; 66(3): 368-75, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20880132

RESUMO

BACKGROUND: The use of recombinant lactic acid bacteria (LAB) as vehicles for mucosal delivery of recombinant allergens is an attractive concept for antigen-defined allergy prevention/treatment. Interventions with LAB are of increasing interest early in life when immune programming is initiated. Here, we investigated the effect of neonatal colonization with a recombinant LAB producing the major birch pollen allergen Bet v 1 in a murine model of type I allergy. METHODS: We constructed a recombinant Lactobacillus (L.) plantarum NCIMB8826 strain constitutively producing Bet v 1 to be used for natural mother-to-offspring mono-colonization of germ-free BALB/c mice. Allergen-specific immunomodulatory effects of the colonization on humoral and cellular immune responses were investigated prior and after sensitization to Bet v 1. RESULTS: Mono-colonization with the Bet v 1 producing L. plantarum induced a Th1-biased immune response at the cellular level, evident in IFN-γ production of splenocytes upon stimulation with Bet v 1. After sensitization with Bet v 1 these mice displayed suppressed IL-4 and IL-5 production in spleen and mesenteric lymph node cell cultures as well as decreased allergen-specific antibody responses (IgG1, IgG2a, and IgE) in sera. This suppression was associated with a significant up-regulation of the regulatory marker Foxp3 at the mRNA level in the spleen cells. CONCLUSION: Intervention at birth with a live recombinant L. plantarum producing a clinically relevant allergen reduces experimental allergy and might therefore become an effective strategy for early intervention against the onset of allergic diseases.


Assuntos
Antígenos de Plantas/imunologia , Imunização , Lactobacillus plantarum/genética , Lactobacillus plantarum/imunologia , Linfócitos T Reguladores/imunologia , Células Th1/imunologia , Alérgenos/administração & dosagem , Alérgenos/genética , Alérgenos/imunologia , Animais , Animais Recém-Nascidos , Antígenos de Plantas/genética , Betula/genética , Betula/imunologia , Citocinas/biossíntese , Citocinas/imunologia , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Hipersensibilidade Imediata , Isotipos de Imunoglobulinas/sangue , Isotipos de Imunoglobulinas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Pólen/genética , Pólen/imunologia , Baço/citologia , Baço/imunologia , Células Th2/imunologia
5.
Prague Med Rep ; 112(1): 5-17, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21470494

RESUMO

The aim of the study was to show patients suffering from the coeliac disease, their real gliadin daily intake, offer them very useful information concerning their diet and to find random possible mistakes. The monitoring was carried out within the context of their routine everyday diet regimen. The daily intake of gliadin in the diet was quantified on the basis of gliadin determination in their current daily food. The gluten-free diet was followed for 30 days. The patients were taking regular daily meals, drinks, and sometimes medicines or food supplements. The patients were provided with instructions, survey forms, digital scales, polyethylen bottles and sacks. The patients took out the stipulated amount, which served as a sample of each of their daily meals. The samples included both homemade meals as well as commercial products. The content of gliadin in daily meal was determined by the sandwich ELISA method. The daily gliadin intake was calculated on the base of the reported amount of meals ingested. 1,900 food samples were analyzed within the framework of this study. Several contaminated commercial foods were found; nevertheless this fact did not influence the otherwise satisfactory overall picture of the daily gliadin intake by the patients followed. The results in 14 patients revealed a satisfactory adherence to the gluten-free diet. It was proved that conscientiousness and awareness on the part of coeliac patients, or those taking care of them, is of paramount importance in determining the choice of foods comprising a gluten-free diet.


Assuntos
Doença Celíaca/dietoterapia , Dieta Livre de Glúten , Gliadina/administração & dosagem , Adolescente , Adulto , Anticorpos/sangue , Criança , Pré-Escolar , Feminino , Análise de Alimentos , Gliadina/análise , Gliadina/imunologia , Humanos , Masculino , Cooperação do Paciente , Transglutaminases/imunologia , Adulto Jovem
6.
Clin Exp Immunol ; 160(2): 215-22, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20030668

RESUMO

Calreticulin, upon translocation to the cell surface, plays a critical role in the recognition of tumour cells and in experimentally induced cellular anti-tumour immunity. However, less is known about anti-calreticulin antibodies and their role in malignancies. Using enzyme-linked immunosorbent assay (ELISA), we found immunoglobulin (Ig)A and/or IgG anti-calreticulin antibodies in sera of approximately 63% of patients with hepatocellular carcinoma (HCC), 57% of patients with colorectal adenocarcinoma (CRA) and 47% of patients with pancreatic adenocarcinoma (PACA), while healthy controls, patients with viral hepatitis C and with chronic pancreatitis reached only 2%, 20% and 31% seropositivity, respectively. We found significantly elevated mean levels of IgA anti-calreticulin antibodies (P < 0.001) in patients with HCC (78.7 +/- 52.3 AU, mean +/- standard deviation), PACA (66.5 +/- 30.9 AU) and CRA (61.8 +/- 25.8 AU) when compared to healthy controls (41.4 +/- 19.2 AU). Significantly elevated mean levels of IgG anti-calreticulin antibodies (P < 0.001) were detected in patients with HCC (121.9 +/- 94.2 AU), gall bladder adenocarcinoma (118.4 +/- 80.0 AU) and PACA (88.7 +/- 55.6 AU) when compared to healthy controls (56.7 +/- 22.9 AU). Pepscan analysis revealed a large number of antigenic epitopes of calreticulin recognized by both IgA and IgG antibodies of patients with HCC and PACA, indicating robust systemic immune response. Moreover, significantly elevated levels of antibodies against peptide KGEWKPRQIDNP (P < 0.001) in these patients, tested by ELISA, confirmed the distinct character of antibody reactivity against calreticulin. The high occurrence and specificity of serum anti-calreticulin autoantibodies in the majority of patients with some gastrointestinal malignancies provide the evidence for their possible clinical relevance.


Assuntos
Adenocarcinoma/imunologia , Anticorpos Antineoplásicos/imunologia , Autoanticorpos/imunologia , Autoantígenos/imunologia , Linfócitos B/imunologia , Calreticulina/imunologia , Carcinoma Hepatocelular/imunologia , Neoplasias Colorretais/imunologia , Neoplasias Hepáticas/imunologia , Proteínas de Neoplasias/imunologia , Neoplasias Pancreáticas/imunologia , Adenocarcinoma/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antineoplásicos/sangue , Especificidade de Anticorpos , Autoanticorpos/sangue , Carcinoma Hepatocelular/sangue , Neoplasias Colorretais/sangue , Ensaio de Imunoadsorção Enzimática , Epitopos/imunologia , Feminino , Neoplasias da Vesícula Biliar/sangue , Neoplasias da Vesícula Biliar/imunologia , Hepatite C Crônica/sangue , Hepatite C Crônica/imunologia , Humanos , Imunoglobulina A/sangue , Imunoglobulina A/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Neoplasias Hepáticas/sangue , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/sangue , Pancreatite/sangue , Pancreatite/imunologia , Adulto Jovem
7.
Physiol Res ; 58(1): 101-110, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-18198984

RESUMO

This study was aimed to evaluate the role of commensal Gram-negative bacterium Bacteroides ovatus in murine model of chronic intestinal inflammation. The attempt to induce chronic colitis was done in Bacteroides ovatus-monoassociated, germ-free and conventional mice either in immunocompetent (BALB/c) mice or in mice with severe combined immunodeficiency (SCID), using 2.5 % dextran-sodium sulfate (DSS) in drinking water (7 days DSS, 7 days water, 7 days DSS). Conventional mice developed chronic colitis. Some of germ-free BALB/c and the majority of germ-free SCID mice did not survive the long-term treatment with DSS due to massive bleeding into the intestinal lumen. However, monocolonization of germ-free mice of both strains with Bacteroides ovatus prior to long-term treatment with DSS protected mice from bleeding, development of intestinal inflammation and precocious death. We observed that though DSS-treated Bacteroides ovatus-colonized SCID mice showed minor morphological changes in colon tissue, jejunal brush-border enzyme activities such as gamma-glutamyltranspeptidase, lactase and alkaline phosphatase were significantly reduced in comparison with DSS-untreated Bacteroides ovatus-colonized mice. This modulation of the enterocyte gamma-glutamyltranspeptidase localized to the brush border membrane has been described for the first time. This enzyme is known to reflect an imbalance between pro-oxidant and anti-oxidant mechanisms, which could be involved in protective effects of colonization of germ-free mice with Bacteroides ovatus against DSS injury.


Assuntos
Bacteroides/crescimento & desenvolvimento , Colite/prevenção & controle , Colo/microbiologia , Fosfatase Alcalina/metabolismo , Animais , Doença Crônica , Colite/induzido quimicamente , Colite/enzimologia , Colite/microbiologia , Colite/patologia , Colo/patologia , Sulfato de Dextrana , Modelos Animais de Doenças , Mucosa Intestinal/enzimologia , Jejuno/enzimologia , Lactase/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Microvilosidades/enzimologia , Índice de Gravidade de Doença , Fatores de Tempo , gama-Glutamiltransferase/metabolismo
8.
Clin Exp Immunol ; 153(3): 351-9, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18637103

RESUMO

Refractory coeliac disease (RCD) is a very rare and dangerous form of CD, in which gluten-free diet loses its therapeutic effect and the damage of intestinal mucosa persists. Because of the adherence to the diet, serological markers of CD [immunoglobulin A (IgA) antibodies against gliadin, tissue transglutaminase (tTG) and endomysium] are often missing in RCD patients. We found substantially elevated levels of IgA anti-calreticulin (CRT) antibodies in the sera of almost all RCD patients tested. These sera were negative for IgA antibodies to gliadin and tTG and only some of them showed IgA antibodies to enterocytes. Analysis of patients' IgA reactivity to CRT fragments (quarters and halves) by Western blotting revealed differences in the specificity of IgA antibodies between RCD and CD patients. We therefore used the Pepscan technique with synthetic overlapping decapeptides of CRT to characterize antigenic epitopes recognized by serum IgA antibodies of RCD patients. Employing this method we demonstrated several dominant antigenic epitopes recognized by IgA antibodies of RCD patients on the CRT molecule. Epitope GVTKAAEKQMKD was recognized predominantly by serum IgA of RCD patients. Our results suggest that testing for serum IgA antibodies against CRT and its selected peptide could be a very useful tool in RCD differential diagnosis.


Assuntos
Anticorpos Anti-Idiotípicos/imunologia , Calreticulina/imunologia , Doença Celíaca/imunologia , Imunoglobulina A/imunologia , Idoso , Anticorpos Anti-Idiotípicos/sangue , Western Blotting , Calreticulina/sangue , Doença Celíaca/sangue , Doença Celíaca/diagnóstico , Dieta Livre de Glúten/efeitos adversos , Enterócitos/química , Enterócitos/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Gliadina/sangue , Gliadina/imunologia , Humanos , Imunoglobulina A/sangue , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Transglutaminases/sangue , Transglutaminases/imunologia
9.
Physiol Res ; 67(4): 613-622, 2018 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-29750882

RESUMO

The alcohol-soluble fraction of wheat gluten (gliadins) induces in genetically susceptible individuals immunologically mediated celiac disease (CLD). However, gliadins and related cereal proteins are not unique foodstuff targets of CLD patients´ immune system. Non-gluten wheat alpha-amylase inhibitor 0.19 (AAI 0.19) has been found to be capable of activating human monocyte-derived dendritic cells and inducing pro-inflammatory status in intestinal mucosa of patients with celiac disease (CLD). The possible contribution of this reactivity in incomplete remission of CLD patients on a gluten-free diet (GFD) is matter of contention. In an attempt to characterize the antigenicity of AAI 0.19 in patients with active CLD, patients on a GFD and healthy controls we developed ELISA employing wheat recombinant AAI 0.19. Using this test we revealed a significant (P<0.001) elevation of IgA anti-AAI 0.19 antibodies (Ab) in patients with active CLD (12 out of 30 patients were seropositive) but also in CLD patients on a GFD (15/46), in contrast to healthy controls (2/59). Anti-AAI 0.19 IgG Ab levels were increased (P<0.001) only in patients with active CLD (14/30) in contrast to the controls. Interestingly, the levels of anti-AAI 0.19 IgG Ab were decreased in CLD patients on a GFD (P<0.001, 1/46) compared to the controls (1/59). Notably, 20 out of 30 patients with active CLD were positive either for IgA or for IgG anti-AAI 0.19 Ab. Thus, the majority of CLD patients developed a robust IgA and IgG Ab response against AAI 0.19. These findings may contribute to the broadening of the knowledge about CLD pathogenesis.


Assuntos
Anticorpos Anti-Idiotípicos/sangue , Doença Celíaca/sangue , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Proteínas de Plantas , Adulto , Idoso , Anticorpos Anti-Idiotípicos/imunologia , Doença Celíaca/diagnóstico , Doença Celíaca/imunologia , Estudos de Coortes , Feminino , Humanos , Imunoglobulina A/imunologia , Imunoglobulina G/imunologia , Masculino , Pessoa de Meia-Idade , Proteínas de Plantas/imunologia , Adulto Jovem
11.
Folia Microbiol (Praha) ; 52(6): 618-26, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-18450224

RESUMO

Germ-free immunocompetent (BALB/c) and immunodeficient (SCID) mice were colonized either by E. coli O6K13 or by E. coli strain Nissle 1917 and intestinal inflammation was induced by administering 2.5% dextran sulfate sodium (DSS) in drinking water. Controls were germ-free mice which demonstrated only mild inflammatory changes after induction of an acute intestinal inflammation with DSS as compared with conventional mice in which acute colitis of the colon mucosa similar to human ulcerative colitis is elicited. In mice monocolonized with the nonpathogenic E. coli Nissle 1917 the inflammatory disease did not develop (damage grade 0) while animals monocolonized with uropathogenic E. coli O6K13 exhibited inflammatory changes similar to those elicited in conventionally reared mice (damage grade 3). In the chronic inflammation model, immunocompetent BALB/c mice monocolonized with E. coli Nissle 1917 showed no conspicuous inflammatory changes of the colon mucosa whereas those monocolonized with E. coli O6K13 developed colon inflammation associated with marked infiltration of inflammatory cells. In contrast to germ-free immunodeficient SCID mice that died after application of DSS, the colon mucosa of SCID mice monoassociated with E. coli Nissle 1917 exhibited only moderate inflammatory changes which were less pronounced than changes of colon mucosa of SCID mice monoassociated with E. coli O6K13.


Assuntos
Colite/induzido quimicamente , Colite/microbiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli/crescimento & desenvolvimento , Trato Gastrointestinal/microbiologia , Inflamação/microbiologia , Animais , Colite/imunologia , Colite/patologia , Sulfato de Dextrana , Escherichia coli/patogenicidade , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/patologia , Vida Livre de Germes , Inflamação/imunologia , Inflamação/patologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID
12.
J Endocrinol ; 191(2): 497-503, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17088420

RESUMO

The effect of glucocorticoids is controlled at the pre-receptor level by the activity of 11beta-hydroxysteroid dehydrogenase (11HSD). The isoform 11HSD1 is an NADP+ -dependent oxidoreductase, usually reductase, that amplifies the action of glucocorticoids due to reduction of the biologically inactive 11-oxo derivatives cortisone and 11-dehydrocorticosterone to cortisol and corticosterone. The NAD+ -dependent isoform (11HSD2) is an oxidase that restrains the effect of hormones due to 11beta-oxidation of cortisol and corticosterone to their 11-oxo derivatives. Although the immunosuppressive and anti-inflammatory effects of glucocorticoids are well known, the relationship between inflammation and local metabolism of glucocorticoids is not well understood. In this study, we demonstrated that colitis induced by dextran sulfate sodium modulates colonic 11HSD1. Experimentally induced intestinal inflammation stimulated colonic NADP+ -dependent but not NAD+ -dependent 11HSD activity. Colonic 11HSD1 mRNA was increased, whereas 11HSD2 mRNA was not changed. Additional parallel studies revealed a similar pattern of 11HSD1 mRNA induction in mesenteric lymph nodes and intestinal intraepithelial lymphocytes, but not in spleen and peritoneal macrophages. These data suggest that inflammation modulates local metabolism of glucocorticoid and support the notion that pre-receptor regulation of endogenous corticosteroids might play a role in inflammatory processes.


Assuntos
11-beta-Hidroxiesteroide Desidrogenases/metabolismo , Colite/enzimologia , Colo/enzimologia , RNA Mensageiro/análise , 11-beta-Hidroxiesteroide Desidrogenases/genética , Animais , Sulfato de Dextrana , Ativação Enzimática , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Reação em Cadeia da Polimerase Via Transcriptase Reversa
13.
Folia Microbiol (Praha) ; 51(5): 478-84, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17176771

RESUMO

Our study examined whether repeated preventive oral administration of live probiotic bacterial strains Escherichia coli O83:K24:H31 (Ec O83), Escherichia coli Nissle 1917 O6:K5:H1 (Ec Nis) and Lactobacillus casei DN 114001 (Lc) can protect mice against dextran sodium sulfate (DSS)-induced colitis. A significant decrease in average symptom score was observed in Ec O83-, Ec Nis- and Lc-pretreated group (p < 0.05). Significant differences in body mass loss between Lc pretreated mice with DSS-induced colitis were found when compared with nontreated mice (p < 0.05). PBS pretreated mice had a significantly shorter colon than Ec O83-, Ec Nis- and Lc-pretreated mice (p < 0.05). Administration of Lc significantly decreased the severity of DSS induced histological marks of inflammation (p < 0.05). A significant difference (p < 0.05) was also found in specific IgA level against given probiotic in enteral fluid between colitic mice and healthy mice pretreated with Ec 083 and Ec Nis.


Assuntos
Colite Ulcerativa/prevenção & controle , Colo/microbiologia , Mucosa Intestinal/patologia , Probióticos/farmacologia , Administração Oral , Animais , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/imunologia , Sulfato de Dextrana/efeitos adversos , Modelos Animais de Doenças , Escherichia coli , Histocitoquímica , Imunoglobulina A/análise , Mucosa Intestinal/imunologia , Lacticaseibacillus casei , Camundongos , Camundongos Endogâmicos BALB C
14.
J Leukoc Biol ; 67(3): 312-8, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10733090

RESUMO

Macrophages play an important role in effector mechanisms of various chronic inflammatory diseases. We studied the effect of gluten, the agent inducing celiac disease, and other food antigens on the activation of macrophages. Nitric oxide (NO) and cytokine production were followed as markers of activation, using cultured murine peritoneal macrophages. None of the food antigens tested caused direct inducible nitric oxide synthase (iNOS) activation in macrophages. Unlike other food antigens gluten, gliadin, and their proteolytic fragments significantly enhanced NO production when applied together with interferon-gamma (IFN-gamma), the most efficient being fragments originating from 25- to 45-min peptic digestion. The activation pathway was mediated via direct stimulation of tumor necrosis factor alpha (TNF-alpha) secretion. The NO-enhancing effect was confirmed at the level of iNOS mRNA transcription. In case of sustained local inflammatory reaction connected with increase of IFN-gamma, gluten and its proteolytic fragments may thus elevate NO production. Increased NO level could consequently participate in the development of mucosal lesions in the gut of celiac patients.


Assuntos
Antígenos/imunologia , Citocinas/biossíntese , Alimentos , Glutens/imunologia , Ativação de Macrófagos/imunologia , Macrófagos Peritoneais/imunologia , Óxido Nítrico/metabolismo , Animais , Células Cultivadas , Citocinas/imunologia , Citocinas/metabolismo , Feminino , Gliadina/síntese química , Gliadina/imunologia , Glutens/farmacologia , Interferon gama/imunologia , Interleucina-10/biossíntese , Interleucina-10/metabolismo , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/enzimologia , Macrófagos Peritoneais/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo II , Nitritos/metabolismo , Fragmentos de Peptídeos/síntese química , Fragmentos de Peptídeos/imunologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ativação Transcricional/efeitos dos fármacos , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/metabolismo
15.
Physiol Res ; 64(4): 537-46, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25470519

RESUMO

Impairment of mucosal barrier integrity of small intestine might be causative in immune-mediated gastrointestinal diseases. We tested the markers of epithelial apoptosis - cytokeratin 18 caspase-cleaved fragment (cCK-18), and enterocyte damage - intestinal fatty acid-binding protein (I-FABP) and soluble CD14 (sCD14) in sera of patients with untreated celiac disease (CLD), those on gluten-free diet (CLD-GFD), patients with autoimmune diabetes mellitus (T1D), T1D with insulitis (T1D/INS), and diabetes mellitus type 2 (T2D). We found elevated levels of cCK-18 (P<0.001), I-FABP (P<0.01) and sCD14 (P<0.05) in CLD when compared to healthy controls. However, the levels of cCK-18 (P<0.01) and I-FABP (P<0.01) in CLD-GFD were higher when compared with controls. Interestingly, elevated levels of cCK-18 and I-FABP were found in T2D and T1D (P<0.001), and T1D/INS (P<0.01, P<0.001). Twenty-two out of 43 CLD patients were seropositive for cCK-18, 19/43 for I-FABP and 11/43 for sCD14; 9/30 of T2D patients were positive for cCK-18 and 5/20 of T1D/INS for sCD14, while in controls only 3/41 were positive for cCK-18, 3/41 for I-FABP and 1/41 for sCD14. We documented for the first time seropositivity for sCD14 in CLD and potential usefulness of serum cCK-18 and I-FABP as markers of gut damage in CLD, CLD-GFD, and diabetes.


Assuntos
Doença Celíaca/sangue , Citocinas/sangue , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 2/sangue , Enterócitos/patologia , Hepatopatias/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose , Biomarcadores/sangue , Doença Celíaca/epidemiologia , Doença Celíaca/patologia , Comorbidade , República Tcheca/epidemiologia , Diabetes Mellitus Tipo 1/epidemiologia , Diabetes Mellitus Tipo 1/patologia , Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Mellitus Tipo 2/patologia , Enterócitos/metabolismo , Feminino , Humanos , Hepatopatias/epidemiologia , Hepatopatias/patologia , Masculino , Pessoa de Meia-Idade , Prevalência , Reprodutibilidade dos Testes , Fatores de Risco , Sensibilidade e Especificidade , Adulto Jovem
16.
Immunol Lett ; 13(1-2): 15-8, 1986 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-3530990

RESUMO

Circulating lymphocytes were enumerated in 25 patients with coeliac disease and in 24 healthy donors by immunofluorescent staining using monoclonal antibodies for T cell surface phenotypic markers T11 (mature), T4 (helper) and T8 (suppressor) or polyvalent antisera for surface immunoglobulins (B cells). Proportions of peripheral T and B cells and percentages and ratio of T cell subsets in coeliac disease were not significantly different from those in controls. Individual ratios of helper to suppressor T cells did not correlate with disease activity or with keeping the diet.


Assuntos
Linfócitos B/classificação , Doença Celíaca/sangue , Linfócitos T/classificação , Adulto , Anticorpos Monoclonais , Linfócitos B/imunologia , Feminino , Imunofluorescência , Glutens/imunologia , Glutens/metabolismo , Humanos , Masculino , Linfócitos T/imunologia
17.
Immunol Lett ; 17(4): 335-8, 1988 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3372012

RESUMO

Monoclonal antibodies reactive with gliadin were prepared by fusion of spleen cells isolated from gliadin-immunized BALB/c mice with myeloma cells. The reactivity of mAbs with different preparations of gliadin and their enzymatic digest were measured using ELISA method. The mAb produced by GL 1 clone was shown to react preferentially with alpha-gliadin and its enzymatic digest.


Assuntos
Anticorpos Monoclonais/imunologia , Doença Celíaca/etiologia , Gliadina/imunologia , Proteínas de Plantas/imunologia , Especificidade de Anticorpos , Antígenos/imunologia , Gliadina/efeitos adversos , Gliadina/isolamento & purificação , Humanos
18.
Dev Comp Immunol ; 6(4): 755-64, 1982.
Artigo em Inglês | MEDLINE | ID: mdl-7160516

RESUMO

We studied age-dependent changes in distribution of receptor-ligand complexes in the lateral plain of plasma membrane after binding of six different lectins and anti H-2a and anti Ias antisera to the surface of mouse peritoneal macrophages. Additionally we examined formation of spontaneous rosettes with sheep red blood cells and the phagocytic and pinocytic activities of these macrophages. Macrophages of young and aged mice exhibited a lower mobility of the studied antigens and lectin receptors (determined as a function of patch and cap formation) and a higher rate of formation of spontaneous rosettes. The percentage of Ia antigens bearing macrophages was very low during early ontogenesis; it increased during adulthood and decreased during ageing. The highest percentage of phagocytizing macrophages was found in adult mice, even thought the phagocytic index (i.e., the average quantity of phagocytized particles in a phagocytizing cell) kept increasing during postnatal life. However, the level of pinocytosis was the same in all three groups.


Assuntos
Envelhecimento , Membrana Celular/imunologia , Macrófagos/imunologia , Animais , Antígenos de Superfície , Feminino , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos A , Fagocitose , Pinocitose , Receptores Imunológicos , Receptores Mitogênicos , Formação de Roseta
19.
Immunobiology ; 204(4): 425-33, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11776397

RESUMO

Colonization of the gut with components of commensal microflora profoundly affects the development of the immune system. The aim of the present study was to investigate mucosal and systemic B cell responses during the first few days after intestinal association of colostrum-deprived piglets reared in germ-free (GF) conditions with non-pathogenic Escherichia coli O86. Specific intestinal anti-E. coli antibodies (Ab), among which IgA Ab prevailed, were found 4 days after colonization (72% of standard) and their amount decreased 11 days later reaching 22% of standard. In contrast to mucosal Ab, specific serum Ab remained at the level of GF animals at day 4 (less than 10% of standard) and markedly increased 15 days after colonization (156% of standard). In addition to the occurrence of specific Ab, increased amounts of total immunoglobulins (Ig) of all isotypes were detected in sera and intestinal washings. Using the ELISPOT method an increased number of IgM, IgG and IgA-secreting lymphocytes were found in spleen, mesenteric lymph nodes (MLN) and Peyer's patches (PP) in colonized animals as compared to GF piglets. Contrary to cells from these lymphatic organs, B cells from thymus were not affected by E. coli stimulation. Our results show that at the onset of intestinal colonization, non-pathogenic E. coli specifically and polyclonally stimulate the mucosal and systemic humoral immunity, but relatively soon after stimulation, mucosal-specific responses in gut decreases, indicating the possible beginning of inhibition mechanisms (oral tolerance).


Assuntos
Anticorpos Antibacterianos/imunologia , Infecções por Escherichia coli/imunologia , Imunoglobulinas/imunologia , Intestinos/imunologia , Animais , Anticorpos Antibacterianos/sangue , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Escherichia coli/imunologia , Infecções por Escherichia coli/sangue , Imunoglobulinas/sangue , Mucosa Intestinal/imunologia , Intestinos/microbiologia , Suínos
20.
Vet Immunol Immunopathol ; 87(3-4): 301-6, 2002 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-12072250

RESUMO

Prenatal and early postnatal immune system development has been studied in minipigs. First leukocytes were observed in the yolk sac and fetal liver (FL) on the 17th day of gestation, the majority of them being SWC3(+). The colonization of the thymus (TH) with leukocytes was observed 21 days later. Two waves of fetal TH colonization with pro-T cells were deduced from the frequency of thymocyte subsets. Thymic B cells and immunoglobulin-secreting cells (Ig-SC) were studied by flow cytometry and ELISPOT, respectively. When the total numbers of fetal Ig-SC were compared, the TH was identified as the main source of natural antibodies and the only site of IgA and IgG synthesis. In germ-free animals, the TH also represented the major site of IgG and IgA production and the number of Ig-SC was not influenced by colonization with microflora. FL and bone marrow were identified as primary B lymphopoietic sites. The phenotype of B precursors was characterized and pre-B II cells were shown to be the dominant mononuclear fraction between DG50 and DG105. In the periphery, relative proportions of lymphocyte subsets were determined. Studies in gnotobiotic piglets have revealed that the appearance of CD4(+)CD8(+) T cells and CD2(-) B cells is absolutely dependent on the contact of immune system with live viruses and bacteria, respectively.


Assuntos
Animais Recém-Nascidos/imunologia , Feto/imunologia , Suínos/imunologia , Animais , Células Produtoras de Anticorpos/fisiologia , Linfócitos B/fisiologia , Vida Livre de Germes , Hematopoese , Linfócitos T/fisiologia
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