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1.
Transplantation ; 103(10): 2075-2089, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31343575

RESUMO

BACKGROUND: Stromal laminins α4 and α5 are differentially regulated in transplant tolerance and immunity, respectively, resulting in altered T-cell trafficking. We hypothesized that laminins directly regulated T-cell activation and polarization. METHODS: Human and mouse CD4 T cells were activated in Th1, Th2, Th17, or regulatory T cell (Treg) environments with/without laminin α4 and/or α5. Laminin α5 receptors were blocked with anti-α6 integrin or anti-α-dystroglycan (αDG) monoclonal antibodies, and T-cell polarization was determined. T-cell receptor transgenic TEa CD4 cells that recognized donor alloantigen were transferred into C57BL/6 mice that received alloantigen or cardiac allografts. Laminin receptors were blocked, and TEa T-cell migration and differentiation were assessed. Laminin expression was measured in several models of immunity and tolerance. RESULTS: In diverse models, laminins α4 and α5 were differentially regulated. Immunity was associated with decreased laminin α4:α5 ratio, while tolerance was associated with an increased ratio. Laminin α4 inhibited CD4+ T-cell proliferation and Th1, Th2, and Th17 polarization but favored Treg induction. Laminin α5 favored T-cell activation and Th1, Th2, and Th17 polarization and inhibited Treg. Laminin α5 was recognized by T cell integrin α6 and is important for activation and inhibition of Treg. Laminin α5 was also recognized by T cell α-DG and required for Th17 differentiation. Anti-α6 integrin or anti-DG prolonged allograft survival. CONCLUSIONS: Laminins α4 and α5 are coinhibitory and costimulatory ligands for human and mouse CD4 T cells, respectively. Laminins and their receptors modulate immune responses by acting as one of the molecular switches for immunity or suppression.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Laminina/metabolismo , Linfonodos/metabolismo , Transferência Adotiva , Animais , Linhagem Celular Tumoral/transplante , Neoplasias do Colo/imunologia , Neoplasias do Colo/patologia , Células Dendríticas/imunologia , Modelos Animais de Doenças , Rejeição de Enxerto/imunologia , Transplante de Coração/efeitos adversos , Humanos , Tolerância Imunológica , Laminina/imunologia , Linfonodos/imunologia , Ativação Linfocitária , Camundongos , Receptores de Antígenos de Linfócitos T/genética , Transdução de Sinais/imunologia
2.
J Biol Chem ; 292(26): 10801-10812, 2017 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-28490634

RESUMO

Compromised gastrointestinal barrier function is strongly associated with the progressive and destructive pathologies of the two main forms of irritable bowel disease (IBD), ulcerative colitis (UC), and Crohn's disease (CD). Matriptase is a membrane-anchored serine protease encoded by suppression of tumorigenicity-14 (ST14) gene, which is critical for epithelial barrier development and homeostasis. Matriptase barrier-protective activity is linked with the glycosylphosphatidylinositol (GPI)-anchored serine protease prostasin, which is a co-factor for matriptase zymogen activation. Here we show that mRNA and protein expression of both matriptase and prostasin are rapidly down-regulated in the initiating inflammatory phases of dextran sulfate sodium (DSS)-induced experimental colitis in mice, and, significantly, the loss of these proteases precedes the appearance of clinical symptoms, suggesting their loss may contribute to disease susceptibility. We used heterozygous St14 hypomorphic mice expressing a promoter-linked ß-gal reporter to show that inflammatory colitis suppresses the activity of the St14 gene promoter. Studies in colonic T84 cell monolayers revealed that barrier disruption by the colitis-associated Th2-type cytokines, IL-4 and IL-13, down-regulates matriptase as well as prostasin through phosphorylation of the transcriptional regulator STAT6 and that inhibition of STAT6 with suberoylanilide hydroxamic acid (SAHA) restores protease expression and reverses cytokine-induced barrier dysfunction. Both matriptase and prostasin are significantly down-regulated in colonic tissues from human subjects with active ulcerative colitis or Crohn's disease, implicating the loss of this barrier-protective protease pathway in the pathogenesis of irritable bowel disease.


Assuntos
Colite Ulcerativa/metabolismo , Doença de Crohn/metabolismo , Interleucina-13/metabolismo , Interleucina-4/metabolismo , Serina Endopeptidases/metabolismo , Animais , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/genética , Colite Ulcerativa/patologia , Colo/metabolismo , Colo/patologia , Doença de Crohn/induzido quimicamente , Doença de Crohn/genética , Doença de Crohn/patologia , Sulfato de Dextrana , Modelos Animais de Doenças , Humanos , Ácidos Hidroxâmicos/farmacologia , Interleucina-13/genética , Interleucina-4/genética , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Mutantes , Fator de Transcrição STAT6/genética , Fator de Transcrição STAT6/metabolismo , Serina Endopeptidases/genética , Vorinostat
3.
Radiat Res ; 187(2): 241-250, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28118112

RESUMO

Detonation of a 10-kiloton nuclear bomb in an urban setting would result in >1 million casualties, the majority of which would present with combined injuries. Combined injuries, such as peripheral tissue trauma and radiation exposure, trigger inflammatory events that lead to multiple organ dysfunction (MOD) and death, with gastrointestinal (GI) and pulmonary involvement playing crucial roles. The objective of this study was to develop an animal model of combined injuries, peripheral tissue trauma (TBX animal model) combined with total body irradiation with 5% bone marrow shielding (TBI/BM5) to investigate if peripheral tissue trauma contributes to reduced survival. Male C57BL/6J mice were exposed to TBX10%, irradiation (TBI/BM5), or combined injuries (TBX10% + TBI/BM5). Experiments were conducted to evaluate mortality at day 7 after TBI/BM5. Serial euthanasia was performed at day 1, 3 and 6 or 7 after TBI/BM5 to evaluate the time course of pathophysiologic processes in combined injuries. Functional tests were performed to assess pulmonary function and GI motility. Postmortem samples of lungs and jejunum were collected to assess tissue damage. Results indicated higher lethality and shorter survival in the TBX10% +T BI/BM5 group than in the TBX10% or TBI/BM5 groups (day 1 vs. day 7 and 6, respectively). TBI/BM5 alone had no effects on the lungs but significantly impaired GI function at day 6. As expected, in the animals that received severe trauma (TBX10%), we observed impairment in lung function and delay in GI transit in the first 3 days, effects that decreased at later time points. Trauma combined with radiation (TBX10% + TBI/BM5) significantly augmented impairment of the lung and GI function in comparison to TBX10% and TBI/BM5 groups at 24 h. Histologic evaluation indicated that combined injuries caused greater tissue damage in the intestines in TBX10% + TBI/BM5 group when compared to other groups. We describe here the first combined tissue trauma/radiation injury model that will allow conduction of mechanistic studies to identify new therapeutic targets and serve as a platform for testing novel therapeutic interventions.


Assuntos
Armas Nucleares , Ferimentos e Lesões , Animais , Modelos Animais de Doenças , Trato Gastrointestinal/fisiopatologia , Trato Gastrointestinal/efeitos da radiação , Pulmão/fisiopatologia , Pulmão/efeitos da radiação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Análise de Sobrevida , Ferimentos e Lesões/fisiopatologia
4.
Handb Exp Pharmacol ; 239: 247-267, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28035531

RESUMO

Neuroimmune communications are facilitated by the production of neurotransmitters by immune cells and the generation of immune mediators by immune cells, which form a functional entity called the "neuroimmune synapse." There are several mechanisms that further facilitate neuroimmune interactions including the anatomic proximity between immune cells and nerves, the expression of receptors for neurotransmitters on immune cells and for immune mediators on nerves, and the receptor-mediated activation of intracellular signaling pathways that modulate nerve and immune phenotype and function. The bidirectional communication between nerves and immune cells is implicated in allostasis, a process that describes the continuous adaptation to an ever-changing environment. Neuroimmune interactions are amplified during inflammation by the influx of activated immune cells that significantly alter the microenvironment. In this context, the types of neurotransmitters released by activated neurons or immune cells can exert pro- or anti-inflammatory effects. Dysregulation of the enteric nervous system control of gastrointestinal functions, such as epithelial permeability and secretion as well as smooth muscle contractility, also contribute to the chronicity of inflammation. Persistent active inflammation in the gut leads to neuroimmune plasticity, which is a structural and functional remodeling in both the neural and immune systems. The importance of neuroimmune interactions has made them an emerging target in the development of novel therapies for GI pathologies.


Assuntos
Sistema Nervoso Entérico/imunologia , Sistema Nervoso Entérico/fisiopatologia , Gastroenteropatias/imunologia , Gastroenteropatias/fisiopatologia , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/inervação , Sistema Imunitário/imunologia , Sistema Imunitário/fisiopatologia , Neuroimunomodulação , Animais , Microambiente Celular , Gastroenteropatias/terapia , Humanos , Imunidade nas Mucosas , Sinapses Imunológicas , Inflamação/imunologia , Inflamação/fisiopatologia , Macrófagos/imunologia , Mastócitos/imunologia , Neuroglia/imunologia , Plasticidade Neuronal , Reflexo , Linfócitos T/imunologia
5.
Am J Physiol Gastrointest Liver Physiol ; 311(1): G130-41, 2016 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-27173511

RESUMO

Despite increased appreciation for the role of nicotinic receptors in the modulation of and response to inflammation, the contribution of muscarinic receptors to mucosal homeostasis, clearance of enteric pathogens, and modulation of immune cell function remains relatively undefined. Uninfected and Nippostrongylus brasiliensis-infected wild-type and type 3 muscarinic receptor (M3R)-deficient (Chrm3(-/-)) mice were studied to determine the contribution of M3R to mucosal homeostasis as well as host defense against the TH2-eliciting enteric nematode N. brasiliensis Intestinal permeability and expression of TH1/TH17 cytokines were increased in uninfected Chrm3(-/-) small intestine. Notably, in Chrm3(-/-) mice infected with N. brasiliensis, small intestinal upregulation of TH2 cytokines was attenuated and nematode clearance was delayed. In Chrm3(-/-) mice, TH2-dependent changes in small intestinal function including smooth muscle hypercontractility, increased epithelial permeability, decreased epithelial secretion and absorption, and goblet cell expansion were absent despite N. brasiliensis infection. These findings identify an important role for M3R in host defense and clearance of N. brasiliensis, and support the expanding role of cholinergic muscarinic receptors in maintaining mucosal homeostasis.


Assuntos
Citocinas/metabolismo , Imunidade nas Mucosas , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Nippostrongylus/patogenicidade , Receptor Muscarínico M3/metabolismo , Infecções por Strongylida/metabolismo , Células Th2/metabolismo , Animais , Células Cultivadas , Citocinas/imunologia , Modelos Animais de Doenças , Predisposição Genética para Doença , Homeostase , Interações Hospedeiro-Patógeno , Mucosa Intestinal/imunologia , Mucosa Intestinal/parasitologia , Intestino Delgado/imunologia , Intestino Delgado/parasitologia , Ativação de Macrófagos , Macrófagos/imunologia , Macrófagos/metabolismo , Macrófagos/parasitologia , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Nippostrongylus/imunologia , Fenótipo , Receptor Muscarínico M3/deficiência , Receptor Muscarínico M3/genética , Infecções por Strongylida/genética , Infecções por Strongylida/imunologia , Infecções por Strongylida/parasitologia , Células Th2/imunologia , Células Th2/parasitologia , Fatores de Tempo
6.
Radiat Res ; 185(6): 591-603, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27223826

RESUMO

In this study, nonhuman primates (NHPs) exposed to lethal doses of total body irradiation (TBI) within the gastrointestinal (GI) acute radiation syndrome range, sparing ∼5% of bone marrow (TBI-BM5), were used to evaluate the mechanisms involved in development of the chronic GI syndrome. TBI increased mucosal permeability in the jejunum (12-14 Gy) and proximal colon (13-14 Gy). TBI-BM5 also impaired mucosal barrier function at doses ranging from 10-12.5 Gy in both small intestine and colon. Timed necropsies of NHPs at 6-180 days after 10 Gy TBI-BM5 showed that changes in small intestine preceded those in the colon. Chronic GI syndrome in NHPs is characterized by continued weight loss and intermittent GI syndrome symptoms. There was a long-lasting decrease in jejunal glucose absorption coincident with reduced expression of the sodium-linked glucose transporter. The small intestine and colon showed a modest upregulation of several different pro-inflammatory mediators such as NOS-2. The persistent inflammation in the post-TBI-BM5 period was associated with a long-lasting impairment of mucosal restitution and a reduced expression of intestinal and serum levels of alkaline phosphatase (ALP). Mucosal healing in the postirradiation period is dependent on sparing of stem cell crypts and maturation of crypt cells into appropriate phenotypes. At 30 days after 10 Gy TBI-BM5, there was a significant downregulation in the gene and protein expression of the stem cell marker Lgr5 but no change in the gene expression of enterocyte or enteroendocrine lineage markers. These data indicate that even a threshold dose of 10 Gy TBI-BM5 induces a persistent impairment of both mucosal barrier function and restitution in the GI tract and that ALP may serve as a biomarker for these events. These findings have important therapeutic implications for the design of medical countermeasures.


Assuntos
Medula Óssea , Trato Gastrointestinal/efeitos da radiação , Lesões Experimentais por Radiação/etiologia , Proteção Radiológica , Irradiação Corporal Total/efeitos adversos , Animais , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/patologia , Trato Gastrointestinal/fisiopatologia , Mediadores da Inflamação/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/efeitos da radiação , Macaca mulatta , Masculino , Lesões Experimentais por Radiação/metabolismo , Lesões Experimentais por Radiação/patologia , Lesões Experimentais por Radiação/fisiopatologia , Regeneração/efeitos da radiação
7.
Inflamm Bowel Dis ; 21(8): 1860-71, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25985244

RESUMO

BACKGROUND: The role of muscarinic receptors in mucosal homeostasis, response to enteric pathogens, and modulation of immune cell function is undefined. METHODS: The contribution of type 3 muscarinic receptors (M3R) to mucosal homeostasis within the colon and host defense against Citrobacter rodentium was determined in uninfected and C. rodentium-infected WT and M3R-deficient (Chrm3) mice. In addition, WT and Chrm3 bone marrow-derived macrophages were studied to determine the ability of M3R to modulate macrophage phenotype and function. RESULTS: In Chrm3 mice, clearance of C. rodentium was delayed despite an amplified TH1/TH17 response. Delayed clearance of C. rodentium from Chrm3 mice was associated with prolonged adherence of bacteria to colonic mucosa, decreased goblet cell number, and decreased mucin 2 gene expression. Treatment of bone marrow-derived macrophages with bethanechol, a muscarinic-selective agonist, induced a classically activated macrophage phenotype, which was dependent on M3R expression. Chrm3 bone marrow-derived macrophages retained their ability to attain a classically activated macrophage phenotype when treated with the TH1 cytokine IFN-γ. CONCLUSIONS: In Chrm3 mice, mucin production is attenuated and is associated with prolonged adherence of C. rodentium to colonic mucosa. The immune response, as characterized by production of TH1/TH17 cytokines, in C. rodentium-infected Chrm3 mice is intact. In addition, M3R activity promotes the development of classically activated macrophages. Our data establish a role for M3R in host defense against C. rodentium through effects on goblet cell mucus production and in the modulation of macrophage phenotype and function.


Assuntos
Citrobacter rodentium/imunologia , Infecções por Enterobacteriaceae/microbiologia , Infecções por Enterobacteriaceae/prevenção & controle , Células Caliciformes/imunologia , Macrófagos/imunologia , Receptor Muscarínico M3/fisiologia , Animais , Proliferação de Células , Citocinas/genética , Citocinas/metabolismo , Infecções por Enterobacteriaceae/imunologia , Feminino , Imunofluorescência , Células Caliciformes/microbiologia , Células Caliciformes/patologia , Macrófagos/microbiologia , Macrófagos/patologia , Masculino , Camundongos , Camundongos Knockout , Mucinas/genética , Mucinas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
8.
J Crohns Colitis ; 9(6): 463-76, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25820018

RESUMO

BACKGROUND AND AIMS: Mast cells [MCs] are implicated in epithelial barrier alterations that characterize inflammatory and functional bowel disorders. In this study, we describe mast cell proteinases [chymases and tryptases] and tight junction [TJ] proteins kinetics in a rat model of postinfectious gut dysfunction. METHODS: Jejunal tissues of control and -infected rats were used. Inflammation-related changes in MCs and the expression of TJ-related proteins were evaluated by immunostaining and reverse transcription-quantitative polymerase chain reaction. Epithelial barrier function was assessed in vitro (Ussing chambers) and in vivo. RESULTS: After infection, intestinal inflammation was associated with a generalized overexpression of MC chymases, peaking between Days 6 and 14. Thereafter, a mucosal MC hyperplasia and a late increase in connective tissue MC counts were observed. From Day 2 post-infection, TJ proteins occludin and claudin-3 expression was down-regulated whereas the pore-forming protein claudin-2 was overexpressed. The expression of proglucagon, precursor of the barrier-enhancing factor glucagon-like peptide-2, was reduced. These changes were associated with an increase in epithelial permeability, both in vitro and in vivo. CONCLUSIONS: Proteinases expression and location of mucosal and connective tissue MCs indicate a time-related pattern in the maturation of intestinal MCs following infection. Altered expression of TJ-related proteins is consistent with a loss of epithelial tightness, and provides a molecular mechanism for the enhanced epithelial permeability observed in inflammatory conditions of the gut.


Assuntos
Mucosa Intestinal/patologia , Mucosa Intestinal/fisiopatologia , Mastócitos/enzimologia , Junções Íntimas/metabolismo , Triquinelose/fisiopatologia , Animais , Quimases/metabolismo , Claudina-2/metabolismo , Claudina-3/metabolismo , Hiperplasia/parasitologia , Doenças Inflamatórias Intestinais/fisiopatologia , Interleucina-6/metabolismo , Mucosa Intestinal/parasitologia , Jejuno/fisiopatologia , Masculino , Ocludina/metabolismo , Permeabilidade , Proglucagon/metabolismo , Ratos , Ratos Sprague-Dawley , Técnicas de Cultura de Tecidos , Trichinella spiralis , Triptases/metabolismo
9.
Gut Microbes ; 5(2): 254-8, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24637799

RESUMO

Host defense is an orchestrated response involving changes in the expression of receptors and release of mediators from both immune and structural cells. There is a growing recognition of the important role of proteolytic pathways for the protective immune response to enteric pathogens. Enteric nematode infection induces a type 2 immune response with polarization of macrophages toward the alternatively activated phenotype (M2). The Th2 cytokines, IL-4, and IL-13, induce a STAT6-dependent upregulation of the expression of the protease inhibitor, serpinB2, which protects macrophages from apoptosis. M2 are critical to worm clearance and a novel role for serpinB2 is its regulation of the chemokine, CCL2, which is necessary for monocyte and/or macrophage influx into small intestine during infection. There is a growing list of factors including immune (LPS, Th2 cytokines) as well as hormonal (gastrin, 5-HT) that are linked to increased expression of serpinB2. Thus, serpinB2 represents an immune regulated factor that has multiple roles in the intestinal mucosa.


Assuntos
Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/parasitologia , Macrófagos/metabolismo , Macrófagos/parasitologia , Infecções por Nematoides/metabolismo , Animais , Citocinas/metabolismo , Humanos , Monócitos/metabolismo , Fator de Transcrição STAT6/metabolismo , Células Th2/metabolismo
10.
PLoS One ; 9(1): e84763, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24465430

RESUMO

Parasitic enteric nematodes induce a type 2 immune response characterized by increased production of Th2 cytokines, IL-4 and IL-13, and recruitment of alternatively activated macrophages (M2) to the site of infection. Nematode infection is associated with changes in epithelial permeability and inhibition of sodium-linked glucose absorption, but the role of M2 in these effects is unknown. Clodronate-containing liposomes were administered prior to and during nematode infection to deplete macrophages and prevent the development of M2 in response to infection with Nippostrongylus brasiliensis. The inhibition of epithelial glucose absorption that is associated with nematode infection involved a macrophage-dependent reduction in SGLT1 activity, with no change in receptor expression, and a macrophage-independent down-regulation of GLUT2 expression. The reduced transport of glucose into the enterocyte is compensated partially by an up-regulation of the constitutive GLUT1 transporter consistent with stress-induced activation of HIF-1α. Thus, nematode infection results in a "lean" epithelial phenotype that features decreased SGLT1 activity, decreased expression of GLUT2 and an emergent dependence on GLUT1 for glucose uptake into the enterocyte. Macrophages do not play a role in enteric nematode infection-induced changes in epithelial barrier function. There is a greater contribution, however, of paracellular absorption of glucose to supply the energy demands of host resistance. These data provide further evidence of the ability of macrophages to alter glucose metabolism of neighboring cells.


Assuntos
Enterócitos/metabolismo , Macrófagos/imunologia , Nippostrongylus/imunologia , Infecções por Strongylida/imunologia , Animais , Transporte Biológico , Células Cultivadas , Ácido Clodrônico/administração & dosagem , Ácido Clodrônico/farmacologia , Enterócitos/imunologia , Enterócitos/parasitologia , Feminino , Expressão Gênica , Glucose/metabolismo , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Transportador de Glucose Tipo 2/genética , Transportador de Glucose Tipo 2/metabolismo , Imunidade Celular , Imunossupressores/administração & dosagem , Imunossupressores/farmacologia , Ativação de Macrófagos , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Transporte Proteico , Infecções por Strongylida/metabolismo , Regulação para Cima/imunologia
11.
J Immunol ; 190(11): 5779-87, 2013 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-23630350

RESUMO

SerpinB2, a member of the serine protease inhibitor family, is expressed by macrophages and is significantly upregulated by inflammation. Recent studies implicated a role for SerpinB2 in the control of Th1 and Th2 immune responses, but the mechanisms of these effects are unknown. In this study, we used mice deficient in SerpinB2 (SerpinB2(-/-)) to investigate its role in the host response to the enteric nematode, Heligmosomoides bakeri. Nematode infection induced a STAT6-dependent increase in intestinal SerpinB2 expression. The H. bakeri-induced upregulation of IL-4 and IL-13 expression was attenuated in SerpinB2(-/-) mice coincident with an impaired worm clearance. In addition, lack of SerpinB2 in mice resulted in a loss of the H. bakeri-induced smooth muscle hypercontractility and a significant delay in infection-induced increase in mucosal permeability. Th2 immunity is generally linked to a CCL2-mediated increase in the infiltration of macrophages that develop into the alternatively activated phenotype (M2). In H. bakeri-infected SerpinB2(-/-) mice, there was an impaired infiltration and alternative activation of macrophages accompanied by a decrease in the intestinal CCL2 expression. Studies in macrophages isolated from SerpinB2(-/-) mice showed a reduced CCL2 expression, but normal M2 development, in response to stimulation of Th2 cytokines. These data demonstrate that the immune regulation of SerpinB2 expression plays a critical role in the development of Th2-mediated protective immunity against nematode infection by a mechanism involving CCL2 production and macrophage infiltration.


Assuntos
Mucosa Intestinal/metabolismo , Intestinos/imunologia , Infecções por Nematoides/imunologia , Infecções por Nematoides/metabolismo , Inibidor 2 de Ativador de Plasminogênio/metabolismo , Células Th2/imunologia , Células Th2/metabolismo , Animais , Citocinas/imunologia , Citocinas/metabolismo , Regulação da Expressão Gênica , Mucosa Intestinal/imunologia , Mucosa Intestinal/parasitologia , Intestinos/parasitologia , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Camundongos Knockout , Monócitos/imunologia , Monócitos/metabolismo , Músculo Liso/metabolismo , Músculo Liso/parasitologia , Infecções por Nematoides/genética , Inibidor 2 de Ativador de Plasminogênio/deficiência , Inibidor 2 de Ativador de Plasminogênio/genética
12.
Infect Immun ; 81(7): 2546-53, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23649095

RESUMO

Heligmosomoides bakeri is a nematode with parasitic development exclusively in the small intestine of infected mice that induces a potent STAT6-dependent Th2 immune response. We previously demonstrated that host protective expulsion of adult H. bakeri worms from a challenge infection was delayed in selenium (Se)-deficient mice. In order to explore mechanisms associated with the delayed expulsion, 3-week-old female BALB/c mice were placed on a torula yeast-based diet with or without 0.2 ppm Se, and after 5 weeks, they were inoculated with H. bakeri infective third-stage larvae (L3s). Two weeks after inoculation, the mice were treated with an anthelmintic and then rested, reinoculated with L3s, and evaluated at various times after reinoculation. Analysis of gene expression in parasite-induced cysts and surrounding tissue isolated from the intestine of infected mice showed that the local-tissue Th2 response was decreased in Se-deficient mice compared to that in Se-adequate mice. In addition, adult worms recovered from Se-deficient mice had higher ATP levels than worms from Se-adequate mice, indicating greater metabolic activity in the face of a suboptimal Se-dependent local immune response. Notably, the process of worm expulsion was restored within 2 to 4 days after feeding a Se-adequate diet to Se-deficient mice. Expulsion was associated with an increased local expression of Th2-associated genes in the small intestine, intestinal glutathione peroxidase activity, secreted Relm-ß protein, anti-H. bakeri IgG1 production, and reduced worm fecundity and ATP-dependent metabolic activity.


Assuntos
Heligmosomatoidea/imunologia , Imunidade Celular/efeitos dos fármacos , Selênio/deficiência , Infecções por Strongylida/imunologia , Trifosfato de Adenosina/metabolismo , Animais , Anti-Helmínticos/farmacologia , Ativação Enzimática , Feminino , Fertilidade , Regulação da Expressão Gênica/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Heligmosomatoidea/patogenicidade , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/imunologia , Larva/imunologia , Larva/patogenicidade , Camundongos , Camundongos Endogâmicos BALB C , Selênio/farmacologia , Infecções por Strongylida/parasitologia , Células Th2/citologia , Células Th2/imunologia , Fatores de Tempo
13.
PLoS One ; 8(3): e59441, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23536877

RESUMO

Type 2 immunity is essential for host protection against nematode infection but is detrimental in allergic inflammation or asthma. There is a major research focus on the effector molecules and specific cell types involved in the initiation of type 2 immunity. Recent work has implicated an important role of epithelial-derived cytokines, IL-25 and IL-33, acting on innate immune cells that are believed to be the initial sources of type 2 cytokines IL-4/IL-5/IL-13. The identities of the cell types that mediate the effects of IL-25/IL-33, however, remain to be fully elucidated. In the present study, we demonstrate that macrophages as IL-25/IL-33-responsive cells play an important role in inducing type 2 immunity using both in vitro and in vivo approaches. Macrophages produced type 2 cytokines IL-5 and IL-13 in response to the stimulation of IL-25/IL-33 in vitro, or were the IL-13-producing cells in mice administrated with exogenous IL-33 or infected with Heligmosomoides bakeri. In addition, IL-33 induced alternative activation of macrophages primarily through autocrine IL-13 activating the IL-4Rα-STAT6 pathway. Moreover, depletion of macrophages attenuated the IL-25/IL-33-induced type 2 immunity in mice, while adoptive transfer of IL-33-activated macrophages into mice with a chronic Heligmosomoides bakeri infection induced worm expulsion accompanied by a potent type 2 protective immune response. Thus, macrophages represent a unique population of the innate immune cells pivotal to type 2 immunity and a potential therapeutic target in controlling type 2 immunity-mediated inflammatory pathologies.


Assuntos
Interleucinas/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Transferência Adotiva , Animais , Citocinas/biossíntese , Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-13/biossíntese , Interleucina-33 , Interleucinas/administração & dosagem , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Infecções por Nematoides/imunologia , Infecções por Nematoides/prevenção & controle
14.
Biochim Biophys Acta ; 1830(2): 2410-26, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22922290

RESUMO

BACKGROUND: The discovery of markers to identify the intestinal stem cell population and the generation of powerful transgenic mouse models to study stem cell physiology have led to seminal discoveries in stem cell biology. SCOPE OF REVIEW: In this review we give an overview of the current knowledge in the field of intestinal stem cells (ISCs) highlighting the most recent progress on markers defining the ISC population and pathways governing intestinal stem cell maintenance and differentiation. Furthermore we review their interaction with other stem cell related pathways. Finally we give an overview of alteration of these pathways in human inflammatory gastrointestinal diseases. MAJOR CONCLUSIONS: We highlight the complex network of interactions occurring among different pathways and put in perspective the many layers of regulation that occur in maintaining the intestinal homeostasis. GENERAL SIGNIFICANCE: Understanding the involvement of ISCs in inflammatory diseases can potentially lead to new therapeutic approaches to treat inflammatory GI pathologies such as IBD and celiac disease and could reveal the molecular mechanisms leading to the pathogenesis of dysplasia and cancer in inflammatory chronic conditions. This article is part of a Special Issue entitled Biochemistry of Stem Cells.


Assuntos
Mucosa Intestinal/metabolismo , Transdução de Sinais , Células-Tronco/metabolismo , Animais , Intestinos/citologia , Camundongos , Células-Tronco/citologia
15.
Health Phys ; 103(4): 400-10, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23091877

RESUMO

The acute effects of irradiation on the gastrointestinal (GI) system are well documented, but the longer-term effects are less well known. Increased incidence of adenocarcinoma has been noted, but apart from descriptions of fibrosis, the development of other pathologies specific to survivors of acute radiation is poorly understood. Samples were taken from C57BL/6 mice irradiated with partial-body irradiation where the thorax, head, and forelimbs were shielded (i.e., sparing 40% of the bone marrow). Tissue from age-matched controls was also collected. There were clear pathological changes in the intestine associated with DEARE (Delayed Effects of Acute Radiation Exposure) at doses greater than 12 Gy, with a dose-related increase in observed pathologies. Mice maintained on the synthetic antibiotic ciprofloxacin during the acute phase (days 4 to 20), however, had a lower or delayed incidence of symptoms. After 20 d, mice developed structures similar to early adenomas. Abnormally high levels of apoptotic and mitotic cells were present in some crypts, along with the early adenomas, suggesting tissue regeneration and areas of deregulated cell turnover. Over time, there was inhibited crypt cell proliferation in animals with advanced symptoms, a blunting of the crypts and villi, and an enlargement of villus girth, with an increasingly acellular and fibrotic extracellular matrix (a characteristic that has been demonstrated previously in aging mice). Together these changes may lead to a reduced functional surface area and less motile intestine. These observations are similar to those seen in geriatric animals, suggesting a premature aging of the GI tract.


Assuntos
Síndrome Aguda da Radiação/etiologia , Síndrome Aguda da Radiação/patologia , Gastroenteropatias/etiologia , Gastroenteropatias/patologia , Lesões Experimentais por Radiação/etiologia , Lesões Experimentais por Radiação/patologia , Irradiação Corporal Total/efeitos adversos , Animais , Relação Dose-Resposta à Radiação , Medicina Baseada em Evidências , Humanos , Dose Letal Mediana , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Doses de Radiação , Análise de Sobrevida , Taxa de Sobrevida , Fatores de Tempo
16.
Immunotherapy ; 4(9): 883-98, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23046232

RESUMO

Crohn's disease and ulcerative colitis are chronic, relapsing inflammatory disorders of the GI tract. In both Crohn's disease and ulcerative colitis, leukocytic infiltration of the mucosa is associated with epithelial damage. Recently, monoclonal antibodies directed against cell adhesion molecules (CAMs) involved in leukocyte extravasation have been developed. Natalizumab, the first drug brought to market targeting CAMs, is clinically effective but is associated with serious adverse effects including the uncommon, but often fatal, neurological disease progressive multifocal leukoencephalopathy. Vedolizumab targets a subset of the CAMs blocked by natalizumab and is currently in Phase III trials to study its efficacy and safety in patients with inflammatory bowel disease. Here, we discuss the current treatment options available for patients with Crohn's disease or ulcerative colitis, the history of CAM inhibitors, the current state of development of vedolizumab and its future role in inflammatory bowel disease, if approved by regulatory agencies.


Assuntos
Anticorpos Monoclonais Humanizados/uso terapêutico , Colite Ulcerativa/tratamento farmacológico , Doença de Crohn/tratamento farmacológico , Animais , Anticorpos Monoclonais Humanizados/efeitos adversos , Anticorpos Monoclonais Humanizados/farmacocinética , Anticorpos Monoclonais Humanizados/farmacologia , Controle de Medicamentos e Entorpecentes , Humanos
17.
Health Phys ; 103(4): 411-26, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22929470

RESUMO

The development of medical countermeasures against the acute gastrointestinal subsyndrome of the acute radiation syndrome in humans requires well characterized and validated animal models. These models must adhere to the criteria of the U.S. Food and Drug Administration's Animal Rule and consider the natural history and clinical context of the human radiation response and treatment in the nuclear terrorist scenario. The models must define the radiation dose- and time-dependent relationships for mortality and major signs of morbidity, including concurrent damage in other organs, such as the bone marrow, that may contribute to the overall mortality and morbidity. There are no such models of the gastrointestinal syndrome in response to total-body irradiation in the nonhuman primate. Herein, these parameters are defined for the rhesus macaque exposed to potentially lethal doses of radiation and administered medical management. Rhesus macaques (n = 69) were exposed bilaterally to 6 MV linear accelerator-derived photon total body irradiation to midline tissue (thorax) doses ranging from 10.0 to 14.0 Gy at 0.80 Gy min(-1). Following irradiation, all animals were administered supportive care consisting of fluids, anti-emetics, anti-diarrheal medication, antibiotics, blood transfusions, analgesics, and nutrition. The primary endpoint was survival at 15 d post-irradiation. Secondary endpoints included indices of dehydration, diarrhea, weight loss, hematological parameters, cellular histology of the small and large intestine, and mean survival time of decedents. Mortality within the 15-d in vivo study defined the acute gastrointestinal syndrome and provided an LD30/15 of 10.76 Gy, LD50/15 of 11.33 Gy, and an LD70/15 of 11.90 Gy. Intestinal crypt and villus loss were dose- and time-dependent with an apparent nadir 7 d post-irradiation and recovery noted thereafter. Severe myelosuppression and thrombocytopenia were noted in all animals, requiring the administration of antibiotics and blood transfusions. The model defines the dose response relationship and time course of acute gastrointestinal syndrome-induced morbidity and mortality in the rhesus macaque.


Assuntos
Síndrome Aguda da Radiação/etiologia , Gastroenteropatias/etiologia , Lesões Experimentais por Radiação/etiologia , Irradiação Corporal Total/efeitos adversos , Síndrome Aguda da Radiação/fisiopatologia , Animais , Relação Dose-Resposta à Radiação , Gastroenteropatias/fisiopatologia , Humanos , Dose Letal Mediana , Macaca mulatta , Masculino , Doses de Radiação , Lesões Experimentais por Radiação/fisiopatologia , Análise de Sobrevida , Taxa de Sobrevida
18.
Health Phys ; 103(4): 427-53, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22929471

RESUMO

The dose response relationship for the acute gastrointestinal syndrome following total-body irradiation prevents analysis of the full recovery and damage to the gastrointestinal system, since all animals succumb to the subsequent 100% lethal hematopoietic syndrome. A partial-body irradiation model with 5% bone marrow sparing was established to investigate the prolonged effects of high-dose radiation on the gastrointestinal system, as well as the concomitant hematopoietic syndrome and other multi-organ injury including the lung. Herein, cellular and clinical parameters link acute and delayed coincident sequelae to radiation dose and time course post-exposure. Male rhesus Macaca mulatta were exposed to partial-body irradiation with 5% bone marrow (tibiae, ankles, feet) sparing using 6 MV linear accelerator photons at a dose rate of 0.80 Gy min(-1) to midline tissue (thorax) doses in the exposure range of 9.0 to 12.5 Gy. Following irradiation, all animals were monitored for multiple organ-specific parameters for 180 d. Animals were administered medical management including administration of intravenous fluids, antiemetics, prophylactic antibiotics, blood transfusions, antidiarrheals, supplemental nutrition, and analgesics. The primary endpoint was survival at 15, 60, or 180 d post-exposure. Secondary endpoints included evaluation of dehydration, diarrhea, hematologic parameters, respiratory distress, histology of small and large intestine, lung radiographs, and mean survival time of decedents. Dose- and time-dependent mortality defined several organ-specific sequelae, with LD50/15 of 11.95 Gy, LD50/60 of 11.01 Gy, and LD50/180 of 9.73 Gy for respective acute gastrointestinal, combined hematopoietic and gastrointestinal, and multi-organ delayed injury to include the lung. This model allows analysis of concomitant multi-organ sequelae, thus providing a link between acute and delayed radiation effects. Specific and multi-organ medical countermeasures can be assessed for efficacy and interaction during the concomitant evolution of acute and delayed key organ-specific subsyndromes.


Assuntos
Síndrome Aguda da Radiação/etiologia , Gastroenteropatias/etiologia , Células-Tronco Hematopoéticas/efeitos da radiação , Insuficiência de Múltiplos Órgãos/etiologia , Lesões Experimentais por Radiação/etiologia , Irradiação Corporal Total/efeitos adversos , Síndrome Aguda da Radiação/fisiopatologia , Animais , Sobrevivência Celular/efeitos da radiação , Relação Dose-Resposta à Radiação , Gastroenteropatias/fisiopatologia , Células-Tronco Hematopoéticas/patologia , Humanos , Dose Letal Mediana , Macaca mulatta , Masculino , Doses de Radiação , Lesões Experimentais por Radiação/fisiopatologia , Análise de Sobrevida , Taxa de Sobrevida
19.
J Leukoc Biol ; 92(4): 753-64, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22782966

RESUMO

Studies of IL-4 have revealed a wealth of information on the diverse roles of this cytokine in homeostatic regulation and disease pathogenesis. Recent data suggest that instead of simple linear regulatory pathways, IL-4 drives regulation that is full of alternatives. In addition to the well-known dichotomous regulation of Th cell differentiation by IL-4, this cytokine is engaged in several other alternative pathways. Its own production involves alternative mRNA splicing, yielding at least two functional isoforms: full-length IL-4, encoded by the IL-4 gene exons 1-4, and IL-4δ2, encoded by exons 1, 3, and 4. The functional effects of these two isoforms are in some ways similar but in other ways quite distinct. When binding to the surface of target cells, IL-4 may differentially engage two different types of receptors. By acting on macrophages, a cell type critically involved in inflammation, IL-4 induces the so-called alternative macrophage activation. In this review, recent advances in understanding these three IL-4-related branch points--alternative splicing of IL-4, differential receptor engagement by IL-4, and differential regulation of macrophage activation by IL-4--are summarized in light of their contributions to inflammation.


Assuntos
Inflamação/etiologia , Interleucina-4/fisiologia , Animais , Asma/etiologia , Humanos , Interleucina-4/genética , Ativação de Macrófagos , Proteína Tirosina Fosfatase não Receptora Tipo 6/fisiologia , Receptores de Interleucina-13/química , Receptores de Interleucina-13/fisiologia , Receptores de Interleucina-4/química , Receptores de Interleucina-4/fisiologia , Escleroderma Sistêmico/etiologia , Transdução de Sinais , Tuberculose/etiologia
20.
Inflamm Bowel Dis ; 18(7): 1303-14, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22081509

RESUMO

BACKGROUND: Matriptase is a membrane-anchored serine protease encoded by suppression of tumorigenicity-14 (ST14) that is required for epithelial barrier homeostasis. However, its functional role in inflammatory bowel disease (IBD) is unexplored. METHODS: Matriptase expression in control, Crohn's disease, and ulcerative colitis tissue specimens was studied by quantitative polymerase chain reaction (qPCR) and immunostaining. Matriptase function was investigated by subjecting St14 hypomorphic and control littermates to dextran sodium sulfate (DSS)-induced colitis and by siRNA silencing in cultured monolayers. Mice were analyzed for clinical, histological, molecular, and cellular effects. RESULTS: Matriptase protein and ST14 mRNA levels are significantly downregulated in inflamed colonic tissues from Crohn's disease and ulcerative colitis patients. Matriptase-deficient St14 hypomorphic mice administered DSS for 7 days followed by water without DSS for 3 days develop a severe colitis, with only 30% of the St14 hypomorphic mice surviving to day 14, compared with 100% of control littermates. Persistent colitis in surviving St14 hypomorphic mice was associated with sustained cytokine production, an inability to recover barrier integrity, and enhanced claudin-2 expression. Cytokines implicated in barrier disruption during IBD suppress matriptase expression in T84 epithelial monolayers and restoration of matriptase improves barrier integrity in the cytokine-perturbed monolayers. CONCLUSIONS: These data demonstrate a critical role for matriptase in restoring barrier function to injured intestinal mucosa during colitis, which is suppressed by excessive activation of the immune system. Strategies to enhance matriptase-mediated barrier recovery could be important for intervening in the cycle of inflammation associated with IBD.


Assuntos
Colite Ulcerativa/metabolismo , Colite/prevenção & controle , Doença de Crohn/metabolismo , Intestinos/enzimologia , Serina Endopeptidases/metabolismo , Serina Endopeptidases/fisiologia , Animais , Western Blotting , Colite/induzido quimicamente , Colite/patologia , Colite Ulcerativa/genética , Colite Ulcerativa/patologia , Colo/enzimologia , Colo/patologia , Doença de Crohn/genética , Doença de Crohn/patologia , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Impedância Elétrica , Feminino , Humanos , Técnicas Imunoenzimáticas , Intestinos/lesões , Masculino , Proteínas de Membrana , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Serina Endopeptidases/genética
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