RESUMO
Stress seems to affect the onset and evolution of diverse illnesses with an inflammatory substrate. Whether physiological or psychological, stress increases epithelial permeability. In the mucosa of the nasal cavity and upper respiratory tract, the epithelial barrier is regulated in large part by bicellular and tricellular tight junctions (bTJs and tTJs, respectively). The junctional complexes are composed of multiple membrane proteins: claudins, tight-junction-associated MARVEL proteins (TAMs: occludin, tricellulin and marvelD3), and scaffolding proteins such as ZO-1, -2 and -3. The aim of the present study was to examine the possible modification of nasal permeability and TJ protein expression in a mouse model of acute psychological stress (a 4-h immobility session). Serum corticosterone was quantified from plasma samples to verify the onset of stress. Evaluation was made of the relative concentration of key proteins in nasal mucosa by using Western blot, and of changes in permeability by analyzing FITC-Dextran leakage from the nose to the blood. Compared to the control, the stressed group showed a greater epithelial permeability to FITC-Dextran, a reduced expression of occludin and tricellulin, and an elevated expression of ZO-2 and claudin-4. This evidence points to increased paracellular flow of large molecules through an altered structure of tTJs. Apparently, the structure of bTJs remained unchanged. The current findings could provide insights into the relation of stress to the onset/exacerbation of respiratory infections and/or allergies.
Assuntos
Corticosterona/sangue , Mucosa Nasal , Estresse Psicológico/metabolismo , Junções Íntimas , Animais , Dextranos , Fluoresceína-5-Isotiocianato/análogos & derivados , Camundongos , Camundongos Endogâmicos BALB C , Mucosa Nasal/metabolismo , Mucosa Nasal/fisiopatologia , Restrição Física , Estresse Psicológico/sangue , Junções Íntimas/metabolismoRESUMO
The protozoan Entamoeba histolytica is the etiological agent of amoebiasis, which can spread to the liver and form amoebic liver abscesses. Histological studies conducted with resistant and susceptible models of amoebic liver abscesses (ALAs) have established that neutrophils are the first cells to contact invasive amoebae at the lesion site. Myeloperoxidase is the most abundant enzyme secreted by neutrophils. It uses hydrogen peroxide secreted by the same cells to oxidize chloride ions and produce hypochlorous acid, which is the most efficient microbicidal system of neutrophils. In a previous report, our group demonstrated that myeloperoxidase presents amoebicidal activity in vitro. The aim of the current contribution was to analyze in vivo the role of myeloperoxidase in a susceptible (hamsters) and resistant (Balb/c mice) animal models of ALAs. In liver samples of hamsters and mice inoculated intraportally with Entamoeba histolytica trophozoites, the number of neutrophils in ALAs was determined by enzymatic activity. The presence of myeloperoxidase was observed by staining, and its expression and activity were quantified in situ. A significant difference existed between the two animal models in the number of neutrophils and the expression and activity of myeloperoxidase, which may explain the distinct evolution of amoebic liver abscesses. Hamsters and mice were treated with an MPO inhibitor (4-aminobenzoic acid hydrazide). Hamsters treated with ABAH showed no significant differences in the percentage of lesions or in the percentage of amoebae damaged compared with the untreated hamsters. ABAH treated mice versus untreated mice showed larger abscesses and a decreased percentage of damaged amoebae in these lesion at all stages of evolution. Further studies are needed to elucidate the host and amoebic mechanisms involved in the adequate or inadequate activation and modulation of myeloperoxidase.
Assuntos
Entamoeba histolytica/fisiologia , Abscesso Hepático Amebiano/enzimologia , Peroxidase/metabolismo , Animais , Cricetinae , Modelos Animais de Doenças , Resistência à Doença , Interações Hospedeiro-Patógeno , Elastase de Leucócito/metabolismo , Fígado/enzimologia , Fígado/imunologia , Fígado/parasitologia , Masculino , Camundongos Endogâmicos BALB C , Neutrófilos/enzimologiaRESUMO
Quadrulella (Amoebozoa, Arcellinida, Hyalospheniidae) is a genus of testate amoebae with unmistakable morphology, which secretes characteristic square plates to reinforce the test. They are mainly known from fens and freshwater habitats and have never been documented in deserts. We describe a new species, Quadrulella texcalense, from biological soil crusts in the intertropical desert of Tehuacán (state of Puebla, Mexico). Quadrulella texcalense occurred only at altitudes between 2140 and 2221m.a.s.l., together with the bryophyte genera Pseudocrossidium, Weissia, Bryum, Didymodon, Neohyophyla and Aloina. The soil was extremely dry (moisture of 1.97-2.6%), which contrasts sharply with previous reports for the Quadrulella genus. Single cell mitochondrial cytochrome oxidase I (COI) barcoding of thirteen isolated cells showed an important morphological variability despite having all the same COI barcode sequence. Quadrulella texcalense was placed in a tree containing other Hyalsopheniidae, including a newly barcoded South African species, Q. elegans. Q. texcalense unambiguously branched within genus Quadrulella in a compact clade but with a long branch, suggesting accelerated evolution due to a transition towards a new environment and/or under-sampling.
Assuntos
Clima Desértico , Lobosea/classificação , Filogenia , Solo/parasitologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Lobosea/citologia , Lobosea/genética , México , Especificidade da EspécieRESUMO
Stress stimuli affect the immune system responses that occur at mucosal membranes, particularly IgA secretion. It has been suggested that acute stress increases the levels of IgA and that sympathetic innervation plays an important role in this process. We herein explore in a murine model how acute stress affects the Th1/Th2/Treg cytokine balance in NALT, and the possible role of glucocorticoids in this effect. Nine-week-old male CD1 mice were divided into three groups: unstressed (control), stressed (subjected to 4h of immobilization), and stressed after pretreatment with a single dose of the corticosterone receptor antagonist RU-486. The parameters evaluated included plasma corticosterone and epinephrine, IgA levels in nasal fluid (by ELISA), the percentage of CD19+B220+IgA+ lymphocytes and CD138+IgA+ plasma cells, and the mRNA expression of heavy α chain, J chain and pIgR. Moreover, the gene and protein expression of Th1 cytokines (TNFα, IL-2 and INF-γ), Th2 cytokines (IL-4 and IL-5) and Treg cytokines (IL-10 and TGFß) were determined in nasal mucosa. The results show that acute stress generated a shift towards the dominance of an anti-inflammatory immune response (Th2 and Treg cytokines), evidenced by a significant rise in the amount of T cells that produce IL4, IL-5 and IL-10. This immune environment may favor IgA biosynthesis by CD138+IgA+ plasma cells, a process mediated mostly by glucocorticoids.
Assuntos
Citocinas/metabolismo , Imunoglobulina A Secretora/imunologia , Subpopulações de Linfócitos/imunologia , Mucosa Nasal/imunologia , Estresse Fisiológico/imunologia , Animais , Biomarcadores , Citocinas/genética , Epinefrina/sangue , Epinefrina/metabolismo , Expressão Gênica , Glucocorticoides/sangue , Glucocorticoides/metabolismo , Imunoglobulina A Secretora/sangue , Imunofenotipagem , Subpopulações de Linfócitos/metabolismo , Masculino , Camundongos , Mucosa Nasal/metabolismo , Plasmócitos/imunologia , Plasmócitos/metabolismo , Estresse Fisiológico/genéticaRESUMO
Stress stimuli affect the immune system of the mucosa, and in particular IgA secretion. It is well documented that intense psychological and physical stress can increase susceptibility to infection by diverse pathogens in the upper respiratory tract. Our workgroup reported that chronic stress caused by immobilization elicits a decrease in nasal IgA levels in mice. Here, we explore how acute stress (caused by 4h of immobilization) affects IgA secretion in the nasal mucosa, and the possible role of the sympathetic nervous system in this effect. Nine-week-old male CD1 mice were divided into four groups: control, chemical sympathectomy (with 6-OHDA) and treatment with nadolol (5mg/kg) or phentolamine (15mg/kg). All these groups were subdivided into stressed and unstressed animals. The parameters evaluated included plasma corticosterone and epinephrine (only in control groups), SIgA levels (by ELISA) and SIgA expression (by Western Blot) in nasal fluid, percentage of IgA+ plasma cells, and mRNA expression of heavy alpha chain, pIgR, TNFα and TGFß in nasal mucosa. Acute stress reduced the percentage of IgA+ cells while increasing the levels of IgA, the two hormones, and the mRNA expression of heavy alpha chain, pIgR, TNFα and TGFß, which resulted in greater synthesis and transport of IgA. The treatments with 6-OHDA and α- and ß-adrenergic receptor blockers suggest that sympathetic innervation by both types of adrenergic receptors is important for the control of SIgA secretion in nasal mucosa during acute stress. The increase in this parameter depended on the cytokines involved in IgA synthesis and transport.
Assuntos
Catecolaminas/metabolismo , Imunoglobulina A/metabolismo , Mucosa/metabolismo , Estresse Psicológico/metabolismo , Antagonistas Adrenérgicos alfa , Antagonistas Adrenérgicos beta/farmacologia , Animais , Corticosterona , Citocinas/metabolismo , Modelos Animais de Doenças , Imunoglobulina A Secretora/metabolismo , Masculino , Camundongos , Mucosa/efeitos dos fármacos , Nadolol/farmacologia , Oxidopamina/farmacologia , Fentolamina/farmacologia , RNA Mensageiro/metabolismo , Simpatectomia Química , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
A comparison was made of the effects of levamisole, the bacterial fractions of Staphylococcus, and Freund's adjuvant on the immunization of rats with the excretory and secretory antigens of Trichinella spiralis muscle larvae. Wistar rats were immunized with the antigen and a saline solution, levamisole (LV), Staphylococcus (ST), or Freund's adjuvant (FA). After immunization, rats were infected, and the parasite burden at muscular phase was calculated for each group. Levels of IgG1 and IgG2 antibodies, as well as levels of two cytokines, IL-4 and IFN-γ, were evaluated during the immunization and postinfection periods. Differences were found in the kinetics of antibody production between groups (p < 0.01). In all cases, there was reactivity with the main 45-, 50-, and 55-kDa antigens of Trichinella muscle larvae. Immunization with FA and ST enhanced the production of IgG1, but only FA showed a significant increase in the production of IFN-γ (p < 0.01), resulting in 86% protection against the infection. In contrast, only 60-70% protection was attained in the ST and LV groups (p < 0.01). These data support the idea that levamisole and Staphylococcus can be used as adjuvant to enhance the humoral response and, at the same time, demonstrate that IFN-γ could be involved in protection against Trichinella.
Assuntos
Adjuvantes Imunológicos/administração & dosagem , Antígenos de Helmintos/imunologia , Adjuvante de Freund/administração & dosagem , Imunização/métodos , Levamisol/administração & dosagem , Trichinella spiralis/imunologia , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/administração & dosagem , Citocinas/metabolismo , Modelos Animais de Doenças , Carga Parasitária , Ratos , Ratos Wistar , Staphylococcus/imunologia , Triquinelose/imunologia , Triquinelose/parasitologia , Triquinelose/prevenção & controleRESUMO
Here, the effects of neurointermediate (NIL), anterior (AL), and total hypophysectomy (HYPOX) on ileal mucosa cells and gut-associated lymphoid tissue (GALT) are reported. Compared with the sham-operated (SHAM) rats, the villi height and goblet cells numbers were significantly decreased in all groups. Lamina propria area decreased in AL and HYPOX, but not in NIL animals. CD8(+) but not CD4(+) lymphocytes decreased in the HYPOX and NIL groups. Paneth cells did not change, while IgA cells, IgM cells, and secretory IgA were significantly decreased in all groups. NIL but not AL animals lost significant numbers of IgA cells and secretory IgA. In summary, pituitary hormones exert lobe-specific regulatory effects on the gut and on GALT.
Assuntos
Mucosa Intestinal/imunologia , Intestino Delgado/imunologia , Tecido Linfoide/imunologia , Adeno-Hipófise/metabolismo , Adeno-Hipófise Parte Intermédia/metabolismo , Hormônios Hipofisários/metabolismo , Animais , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Células Caliciformes/imunologia , Células Caliciformes/metabolismo , Hormônio do Crescimento/imunologia , Hormônio do Crescimento/metabolismo , Hipofisectomia , Sistema Hipotálamo-Hipofisário/metabolismo , Imunoglobulina A/imunologia , Imunoglobulina A/metabolismo , Imunoglobulina M/imunologia , Imunoglobulina M/metabolismo , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Tecido Linfoide/metabolismo , Masculino , Celulas de Paneth/metabolismo , Adeno-Hipófise/cirurgia , Adeno-Hipófise Parte Intermédia/cirurgia , Neuro-Hipófise/metabolismo , Neuro-Hipófise/cirurgia , Hormônios Hipofisários/imunologia , Sistema Hipófise-Suprarrenal/metabolismo , Prolactina/imunologia , Prolactina/metabolismo , Ratos , Ratos Wistar , Vasopressinas/imunologia , Vasopressinas/metabolismoRESUMO
Although caloric restriction (CR) apparently has beneficial effects on the immune system, its effects on the immunological function of the intestinal mucosa are little known. The present study explored the effect of CR on the innate and adaptive intestinal immunity of mice. Balb/c mice were either fed ad libitum (control) or on alternate days fed ad libitum and fasted (caloric restriction). After 4 months, an evaluation was made of IgA levels in the ileum, the gene expression for IgA and its receptor (pIgR), as well as the expression of two antimicrobial enzymes (lysozyme and phospholipase A2) and several cytokines of the intestinal mucosa. CR increased the gene expression of lysozyme and phospholipase A2. The levels of IgA were diminished in the ileum, which apparently was a consequence of the reduced transport of IgA by pIgR. In ileum, CR increased the gene expression for most cytokines, both pro- and anti-inflammatory. Hence, CR differentially modified the expression of innate and adaptive immunity mediators in the intestine.
Assuntos
Imunidade Adaptativa , Restrição Calórica , Duodeno/imunologia , Íleo/imunologia , Imunidade Inata , Animais , Peso Corporal , Corticosterona/sangue , Citocinas/genética , Citocinas/metabolismo , Duodeno/enzimologia , Perfilação da Expressão Gênica , Íleo/enzimologia , Íleo/metabolismo , Cadeias J de Imunoglobulina/genética , Cadeias J de Imunoglobulina/metabolismo , Cadeias alfa de Imunoglobulina/genética , Cadeias alfa de Imunoglobulina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Muramidase/genética , Muramidase/metabolismo , Norepinefrina/sangue , Fosfolipases A2/genética , Fosfolipases A2/metabolismo , Receptores Fc/genética , Receptores Fc/metabolismo , Transcrição GênicaRESUMO
The aim of the present study was to determine the effect of moderate exercise on the production and secretion of IgA in mouse duodenum, on lymphocyte levels in the lamina propria, and on gene expression encoding for cytokines that regulate the synthesis of α-chain of IgA and the expression of pIgR in the lamina propria. Two groups of young Balb/c mice were fed ad libitum, one sedentary and the other with an exercise program (swimming) for 16 weeks. IgA levels in the duodenum were quantified by ELISA; the number of IgA containing cells as well as B cells, CD4(+) and CD8(+) T cells in the duodenal mucosa was determined by immunohistochemistry; gene expression was analyzed by real-time PCR, and the expression of proteins by Western blotting. Because of physical training, in the duodenum there was a decrease in the number of IgA producing cells, but an increase in the levels of IgA. Additionally, exercise increased the expression of the genes encoding for IL-4, IL-6, IL-10, TNF-α and TGF ß, cytokines that regulate the synthesis of IgA and pIgR, the inflammatory response, and the immune response in the intestine. Thus, the increased IgA found in the duodenal lumen is probably due to the increased production of IgA in the LP and the increased transport of the pIgA-pIgR complex across epithelial cells. Possibly the increased S-IgA levels in the bile also contribute to the change in IgA levels.
Assuntos
Imunoglobulina A/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/citologia , Linfócitos/citologia , Condicionamento Físico Animal/fisiologia , Animais , Linfócitos B/citologia , Linfócitos B/metabolismo , Linfócitos T CD4-Positivos/citologia , Contagem de Células , Corticosterona/sangue , Duodeno/citologia , Duodeno/imunologia , Duodeno/metabolismo , Expressão Gênica/fisiologia , Cadeias J de Imunoglobulina/genética , Cadeias J de Imunoglobulina/metabolismo , Cadeias alfa de Imunoglobulina/genética , Cadeias alfa de Imunoglobulina/metabolismo , Interferon gama/genética , Interleucinas/genética , Intestinos/imunologia , Fígado/metabolismo , Linfócitos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mucosa/citologia , Mucosa/imunologia , Mucosa/metabolismo , Norepinefrina/sangue , Receptores de Imunoglobulina Polimérica/genética , Receptores de Imunoglobulina Polimérica/metabolismo , Natação/fisiologia , Fator de Crescimento Transformador beta/genética , Fator de Necrose Tumoral alfa/genéticaRESUMO
The few reports that have analyzed the effects of stress on the immune cells of the intestinal mucosa or the functions of these cells have tended to focus on S-IgA levels in saliva, and these studies have shown contradictory results. The principal objective of this study was to analyze the effects of repeated restraint stress on the number and distribution of immune cells in Peyer's patches (PPs) as well as the effects of glucocorticoid and catecholamine administration on the same stress-related parameters. Upon analyzing the effect of repeated restraint stress on PPs, it was found that there was no modification in the morphological structure of the PPs but that restraint stress reduced the total number of lymphocytes and the number of CD8+ T cells, B cells, and plasma cells in PPs. Only at the site of PPs where IgA-producing plasma cells are most numerous (the dome) was a decrease found in this type of cell. These effects were due at least in part to the effect of glucocorticoids and catecholamines. Since IgA produced in PPs is a natural antibody that impedes bacterial infections, repeated stress may favor the entry of pathogens through the intestine.
Assuntos
Tolerância Imunológica/imunologia , Imunoglobulina A/biossíntese , Mucosa Intestinal/imunologia , Subpopulações de Linfócitos/imunologia , Nódulos Linfáticos Agregados/imunologia , Estresse Psicológico/imunologia , Animais , Modelos Animais de Doenças , Mucosa Intestinal/citologia , Contagem de Linfócitos , Subpopulações de Linfócitos/citologia , Subpopulações de Linfócitos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Nódulos Linfáticos Agregados/citologia , Restrição Física , Estresse Psicológico/patologiaRESUMO
The effects of stress on the mucosal immune responses in inflammatory disorders of the gut, as well as on salivary and intestinal IgA levels are well known. However, its effects on the structure and function of the NALT have not yet been reported, and are examined in the present study. Balb/c mice were submitted to restraint stress for 3h per day during 4 or 8d. The immunohistochemistry and flow cytometric analysis revealed that repeated restraint stress (4 and 8d) decreased the percentage, compared to the control group, of CD3(+) and CD4(+) T cells, without affecting the percentage of CD8(+) T cells or B220(+) cells (B cells). The numbers of IELs (CD4(+) and CD8(+) T cells) were lower at 4d of stress and higher at 8d. IgA(+) cells in NALT and nasal IgA levels showed a similar pattern, being significantly lower at 4d of stress and significantly higher at 8d. In summary, repeated restraint stress altered the distribution and number of lymphocytes and IgA(+) cells in nasal mucosa, probably due to changes in norepinephrine and corticosterone levels.
Assuntos
Imunidade nas Mucosas/imunologia , Imunoglobulina A/imunologia , Mucosa Nasal/imunologia , Estresse Fisiológico/imunologia , Animais , Antígenos CD/sangue , Antígenos CD/imunologia , Linfócitos B/imunologia , Linfócitos B/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Corticosterona/sangue , Corticosterona/imunologia , Imunoglobulina A/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mucosa Nasal/metabolismo , Norepinefrina/sangue , Norepinefrina/imunologia , Restrição Física , Fatores de TempoRESUMO
The aim of this study was to determine the effect of caloric restriction (CR) in mouse small intestine on the production and secretion of immunoglobulin (Ig) A, the population of lymphocytes in the lamina propria, and the expression of cytokines that mediate and regulate innate and adaptive immunity. One group of young Balb/c mice was fed ad libitum, while the CR group was fed ad libitum and fasted on alternate days. When mice were six months old, IgA levels in the proximal small intestine were quantified by enzyme-linked immunosorbent assay, while the number of IgA containing cells, CD4(+) T cells and CD8(+) T cells in the duodenal mucosa was determined by immunohistochemistry. Furthermore, the expression of several intestinal cytokines, the genes for α-chain IgA, and the polymeric Ig receptor (pIgR) were analyzed by real-time polymerase chain reaction. CR decreased the levels of IgA in the intestine, apparently a consequence of a reduced number of IgA(+) cells in the lamina propria that decrease the production and secretion of this Ig, and a reduced secretion of S-IgA into the bile, which in turn discharges into the proximal intestine. Contrarily, CR increased the expression of genes for α-chain IgA, and the pIgR, indicating that transport of IgA was not a key factor in the decrease of this Ig. Additionally, CR modified the expression of genes for tumor necrosis factor-α, interferon-γ, tumor growth factor-ß, interleukin (IL)-2 and IL-10, all of which regulate the synthesis of IgA and pIgR, the inflammatory response, and the immune response in the intestine.
Assuntos
Restrição Calórica , Citocinas/metabolismo , Imunoglobulina A/metabolismo , Intestino Delgado/imunologia , Animais , Corticosterona/sangue , Citocinas/genética , Ensaio de Imunoadsorção Enzimática , Imunoglobulina A/genética , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-2/genética , Interleucina-2/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Norepinefrina/sangue , RNA Mensageiro/metabolismoRESUMO
Lactoferrin (Lf) is a multifunctional iron-binding glycoprotein with antibacterial and immunomodulatory activities. The antibacterial influence of orally administered bovine Lf (bLf) against murine infection caused by Salmonella typhimurium (S. typhimurium) has scarcely been explored. In the current study, Balb/c mice were treated orally for 7 days with either 5 or 100mg of bovine lactoferrin (bLf). On day 7 of treatment, mice were intragastrically infected with a lethal or sublethal dose of colony forming units (CFU) of S. typhimurium. During treatment with bLf, feces from mice sublethally infected were harvested daily to prepare fecal suspensions, which were serially diluted and plated onto Salmonella Shigella agar to estimate CFU/g of feces. After sacrificing the animals on day 7, 14 or 21 post-infection, samples of intestinal fluid, Peyer's patches (PP), liver and spleen were collected to count the number of CFU by plate dilution. Intestinal secretions were also employed, along with serum samples, to evaluate total IgA, IgG and IgM antibodies, and those against Salmonella surface proteins and bLf by ELISA assay. In lethally infected mice both bLf doses decreased mortality. In sublethally infected mice, both bLf doses decreased bacterial shedding in feces and intestinal fluid, and also reduced bacterial colonization at PP and bacterial translocation in the liver and spleen. Levels of total and those IgG and IgM in serum and IgA in intestinal secretions against Salmonella surface proteins and bLf were enhanced with both doses of bLf. These findings suggest that the effect of bLf against the infection by S. typhimurium in mice may be the result of an antimicrobial activity linked with its modulatory effect on immunocompetent cells (from intestinal and peripheral organs) involved in antibody production.
Assuntos
Anticorpos Antibacterianos/sangue , Lactoferrina/farmacologia , Salmonelose Animal/tratamento farmacológico , Salmonella typhimurium/efeitos dos fármacos , Administração Oral , Animais , Anticorpos Antibacterianos/imunologia , Carga Bacteriana , Translocação Bacteriana/efeitos dos fármacos , Bovinos , Ensaio de Imunoadsorção Enzimática , Fezes/microbiologia , Imunoglobulina A/sangue , Imunoglobulina A/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Intestinos/efeitos dos fármacos , Intestinos/microbiologia , Lactoferrina/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Salmonelose Animal/imunologia , Salmonelose Animal/mortalidade , Salmonella typhimurium/imunologia , Taxa de Sobrevida , Fatores de TempoRESUMO
The most abundant intestinal immunoglobulin and first line of specific immunological defense against environmental antigens is secretory immunoglobulin A. To better understand the effect of repeated stress on the secretion of intestinal IgA, the effects of restraint stress on IgA concentration and mRNA expression of the gene for the alpha-chain of IgA was assessed in both the duodenum and ileum of the rats. Restraint stress induced an increase in intestinal IgA, which was blocked by an adrenalectomy, suggesting a role of catecholamines and glucocorticoids. Whereas the blocking of glucocorticoid receptors by RU-486 did not affect the increased IgA concentration, it did reduce IgA alpha-chain mRNA expression in both segments, indicating a possible mediation on the part of glucocorticoids in IgA secretion by individual cells. Treatment with corticosterone significantly increased both the IgA concentration and IgA alpha-chain mRNA expression in ileum but not in duodenum, suggesting that glucocorticoids may act directly on IgA-antibody forming cells to increase IgA secretion in the former segment. A probable role by catecholamines was evidenced by the reduction in IgA concentration and IgA alpha-chain mRNA expression in both segments after a chemical sympathectomy with 6-hydroxydopamine (6-OHDA). Additionally, norepinephrine significantly reduced IgA alpha-chain mRNA levels but increased pIgR mRNA expression and IgA concentration in both intestinal segments. We propose that the increased intestinal IgA levels caused by repeated restraint stress is likely due to the effects of catecholamines on the transport of plgA across the epithelium.
Assuntos
Imunoglobulina A/metabolismo , Intestino Delgado/metabolismo , Estresse Psicológico/metabolismo , Adrenalectomia , Animais , Catecolaminas/fisiologia , Corticosterona/farmacologia , Duodeno/metabolismo , Ensaio de Imunoadsorção Enzimática , Glucocorticoides/fisiologia , Antagonistas de Hormônios/farmacologia , Íleo/metabolismo , Imunoglobulina A/imunologia , Intestino Delgado/imunologia , Masculino , Mifepristona/farmacologia , Norepinefrina/farmacologia , RNA/biossíntese , RNA/isolamento & purificação , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Wistar , Restrição Física , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estresse Psicológico/imunologia , Simpatectomia QuímicaRESUMO
Few reports exist on the differences in cell populations or immunological functions between the proximal and distal segments of the small intestine (SI). In the current contribution we analyzed the expression of the polymeric immunoglobulin receptor (pIgR) and alpha chains as well as the density of IgA-producing cells from the proximal and distal intestinal segments from Balb/c mice. Furthermore, by using real-time RT-PCR we quantified the expression of cytokines (TNF-alpha, IFN-gamma, IL-4 and TGF-beta), Toll-like receptor-4 (TLR-4), and the glucocorticoid receptor (GR) involved in pIgR expression in intestinal epithelial cells (IEC). In this study, for the first time it has been demonstrated that the expression of the pIgR as well as alpha chain was greater in the proximal than the distal segment of the small intestine of normal mice. Moreover, we found striking differences in the expression of cytokines at the different intestinal compartments. Whereas the expression of TNF-alpha, IFN-gamma and TGF-beta was higher in lamina propria lymphocytes (LPL) of the distal than proximal segment, it was higher in IEC of the proximal than distal segment. In contrast, the expression of the gene for IL-4 was higher in the LPL of the proximal segment and the IEC of the distal segment. Although the overall expression of TNF-alpha, IL-4, IFN-gamma and TGF-beta was higher in the whole mucosa of the distal than proximal segment, we propose that cytokines produced by epithelial cells (TNF-alpha, IFN-gamma and TGF-beta) autocrinally up-regulate the expression of mRNA for the pIgR. Finally the expression of the GR was higher in the proximal segment, while the expression of the gene for TLR-4 was significantly higher in the IEC of the distal than proximal segment. The higher expression of pIgR found in the proximal segment is probably related to the effect on epithelial cells of the higher production of TNF-alpha, IFN-gamma and TGF-beta, as well as the higher expression of the glucocorticoid receptors. The increased expression of pIgR in the proximal segment appears primarily responsible for the increased secretory IgA levels in the small intestine of mice. These results confirm and extend previous findings supporting the compartmentalization of the intestinal immune system.
Assuntos
Imunoglobulina A/metabolismo , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Receptores de Imunoglobulina Polimérica/metabolismo , Animais , Anticorpos , Apoptose , Linfócitos B/imunologia , Linfócitos B/metabolismo , Linfócitos B/patologia , Células Cultivadas , Citocinas/metabolismo , Imunização Passiva , Imunoglobulina A/imunologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Intestino Delgado/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Especificidade de Órgãos , Estrutura Terciária de Proteína/genética , Coelhos , Receptores de Glucocorticoides/metabolismo , Receptores de Imunoglobulina Polimérica/genética , Receptores de Imunoglobulina Polimérica/imunologia , Transdução de Sinais , Baço/imunologia , Baço/metabolismo , Baço/patologiaRESUMO
The two current models of invasive amebiasis both hold that direct contact of toxic molecules and amebas with tissue produces the necrotic areas characteristic of this disorder. Whereas one model characterizes these toxic molecules as amebic products (e.g., lectins, amebapores, cysteine proteinases and other proteolytic enzymes), the other describes them as products of the inflammatory response (e.g., cytokines, nitric oxide, reactive oxygen intermediates and cytotoxic granules). Both these models can account for necrotic areas with many amebas present and with acute inflammation, but not those with few or no amebas present or with scarce inflammation. A new model poses that an inadequate immune response leads to a continuous and prolonged activation of endothelial cells (ECs) by amebas, amebic molecules and cytokines, which triggers the mechanisms leading to necrosis. Other toxic molecules later contribute to EC activation: nitric oxide, reactive oxygen intermediates, the activated complement and proteases. Hyperactivated endothelial cells continuously express adhesion molecules (e.g., ICAM-1 and E-selectin), pro-coagulant molecules (e.g., tissue factor, von Willebrand factor, and the plasminogen activator inhibitor), resulting in ever greater inflammation and thrombosis, which eventually reduces or blocks blood flow in some vessels and starves certain tissue areas of an adequate oxygen and nutrient supply. When necrotic areas first develop, they are surrounded by inflammatory cells due to the acute inflammation at this stage. However, these cells are starved of oxygen and essential nutrients by the same microcirculatory dysfunction. The increasing concentration of nitric oxide during amebiasis eventually has an anti-inflammatory and vasodilating effect, creating a new mechanism for the microcirculatory dysfunction. This local microcirculatory dysfunction can explain necrotic areas in the presence of many, few, or no amebas, with abundant or scarce inflammation.
Assuntos
Amebíase/fisiopatologia , Microcirculação , Animais , Endotélio Vascular/fisiopatologia , Feminino , Humanos , Imunidade Inata , Masculino , Modelos Teóricos , RatosRESUMO
The human large intestine can harbor two morphologically similar amoebae; the invasive Entamoeba histolytica and the non-invasive Entamoeba dispar. Whereas E. histolytica can produce intestinal and extra-intestinal lesions, E. dispar is present in non-symptomatic carriers. Although biochemical, genetic and proteomic studies have identified clear differences between these Entamoebae, it has become clear that several molecules, once assumed to be involved in tissue destruction, exist in both the virulent and the avirulent species. As surface molecules may play a role in invasion and could therefore determine which amoebae are invasive, we analyzed the glycoconjugate composition of E. histolytica and E. dispar using lectins. There was a significant difference between E. histolytica and E. dispar in the expression of glycoconjugates containing d-mannose and N-acetyl-alpha-D-galactosamine residues, but not between virulent and avirulent strains of E. histolytica. N-glycoconjugates with terminal alpha (1-3)-linked mannose residues participate in the adhesion and subsequent cytotoxicity of E. histolytica to cultured hamster hepatocytes. One of them probably is the Gal/GalNAc lectin.
Assuntos
Entamoeba histolytica/metabolismo , Entamoeba/metabolismo , Glicoconjugados/metabolismo , Acetilgalactosamina/análise , Animais , Adesão Celular , Células Cultivadas , Cricetinae , Entamoeba/classificação , Entamoeba/crescimento & desenvolvimento , Entamoeba/patogenicidade , Entamoeba histolytica/crescimento & desenvolvimento , Entamoeba histolytica/patogenicidade , Entamebíase/parasitologia , Entamebíase/patologia , Glicoconjugados/química , Hepatócitos/parasitologia , Hepatócitos/patologia , Abscesso Hepático Amebiano/parasitologia , Abscesso Hepático Amebiano/patologia , Manose/análise , VirulênciaRESUMO
Although innate and adaptive immunity both play a role in amoebiasis, the mechanisms involved in the elimination of Entamoeba histolytica are poorly understood. To provide more information about the innate immune mechanisms that may confer protection against invasive amoebiasis, we administered inflammatory substances (bacillus Calmette-Guérin, lipopolysaccharide, complete Freund's adjuvant, or mineral oil) into the peritoneum of hamsters. The animals were then challenged with pathogenic trophozoites of E. histolytica and, after 7 days, the protective host response was analysed. We found that the nonspecific inflammatory response induced in the peritoneum was sufficient to prevent liver invasion by E. histolytica. In vitro experiments showed that the killing of trophozoites was mediated by peritoneal macrophages and a protein of 68 kDa with peroxidase activity.
Assuntos
Entamoeba histolytica/imunologia , Entamoeba histolytica/patogenicidade , Entamebíase/imunologia , Entamebíase/prevenção & controle , Entamebíase/parasitologia , Imunidade Inata , Animais , Formação de Anticorpos , Cricetinae , Disenteria Amebiana/imunologia , Disenteria Amebiana/parasitologia , Exsudatos e Transudatos/imunologia , Exsudatos e Transudatos/parasitologia , Adjuvante de Freund/imunologia , Humanos , Injeções Intraperitoneais , Abscesso Hepático Amebiano/imunologia , Abscesso Hepático Amebiano/parasitologia , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/parasitologia , Masculino , Óleo Mineral , Mycobacterium bovis/imunologia , Óxido Nítrico/imunologia , Peroxidase/imunologia , Trofozoítos/imunologiaRESUMO
Since the role of striatal GABAergic medium-sized spiny (MSP) neurons in the modulation of the immune responses is largely unknown, we evaluated the humoral immune response in rats with bilateral lesion of the striatum caused by quinolinic acid, which destroys MSP neurons. Sham-operated rats and those with striatal lesions were immunized either with TNP-LPS, a T-independent antigen type 1, or one of several T-dependent antigens: ovoalbumin, bovine serum albumin, lysozyme, sheep red blood cells (SRBC) or outer membrane proteins (OMP) of Salmonella enterica serovar Typhimurium. The specific levels of serum IgM and IgG, as well as intestinal IgA antibodies were determined either by enzyme-linked immunosorbent assay (ELISA) or a haemagglutination assay 5 or 7 days after immunization. Our results show that the lesion of striatal MSP neurons attenuated the primary antibody response to the T-independent antigen type 1 (TNP-LPS), but increased the antibody response to T-dependent antigens (proteins, SRBC and OMP), indicating that the striatal neurons modulate the humoral immune response in rats. The mechanisms involved are probably related to a reduction in both the number of B cells and the expression of caveolin-1 in the spleen, as well as an increase in the number of CD4(+) T cells and in corticosterone levels of the serum.
Assuntos
Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Proteínas da Membrana Bacteriana Externa/farmacologia , Corpo Estriado/imunologia , Lipopolissacarídeos/farmacologia , Salmonella typhimurium/imunologia , Animais , Linfócitos T CD4-Positivos/citologia , Caveolina 1/metabolismo , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/lesões , Corticosterona/sangue , Citocinas/genética , Regulação da Expressão Gênica , Isotipos de Imunoglobulinas/sangue , Isotipos de Imunoglobulinas/imunologia , Masculino , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Baço/imunologiaRESUMO
The impact of restraint stress on the intestinal immune system, particularly on intestinal intraepithelial lymphocytes (i-IEL), has not been described in detail. Thus, the purpose of this study was to assess the effects of restraint stress, including those produced by increases in glucocorticoids and catecholamines, on the population of i-IEL. Mice were exposed to 1 or 4h restraint stress for 4 day, and the number of IEL in the mucosa of the proximal small intestine was determined by immunohistochemistry. The effects of restraint were also analyzed in mice submitted to different procedures: adrenalectomy, chemical sympathectomy, and treatment with a glucocorticoid antagonist (RU486), dexamethasone, and epinephrine. The main findings were that: (1) chronic restraint-stress reduced the i-IEl population in the small intestine; (2) adrenalectomy, treatment with RU-486 and chemical sympathectomy decreased the number of gammadelta, CD4+ and CD8+ T cells in non-stressed groups; (3) dexamethasone reduced the number of gammadelta and CD8+ T cells, and (4) epinephrine reduced the number of gammadelta, CD4+ and CD8+ T cells. These results demonstrated that restraint stress decreased the number of i-IEL in the proximal small intestine of mice, mainly by the combined action of higher concentrations of catecholamines and glucocorticoids, and that lower concentrations of glucocorticoids and catecholamines in unstressed mice preserved the population of i-IEL.