RESUMEN
Activation of Toll-like receptors (TLRs) by pathogens triggers cytokine production and T cell activation, immune defense mechanisms that are linked to immunopathology. Here we show that IFN-γ production by CD4(+) T(H)1 cells during mucosal responses to the protozoan parasite Toxoplasma gondii resulted in dysbiosis and the elimination of Paneth cells. Paneth cell death led to loss of antimicrobial peptides and occurred in conjunction with uncontrolled expansion of the Enterobacteriaceae family of Gram-negative bacteria. The expanded intestinal bacteria were required for the parasite-induced intestinal pathology. The investigation of cell type-specific factors regulating T(H)1 polarization during T. gondii infection identified the T cell-intrinsic TLR pathway as a major regulator of IFN-γ production in CD4(+) T cells responsible for Paneth cell death, dysbiosis and intestinal immunopathology.
Asunto(s)
Infecciones por Enterobacteriaceae/patología , Enterobacteriaceae/crecimiento & desarrollo , Células de Paneth/patología , Transducción de Señal/inmunología , Células TH1/patología , Toxoplasma/crecimiento & desarrollo , Toxoplasmosis Animal/patología , Animales , Linfocitos T CD4-Positivos , Muerte Celular , Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/complicaciones , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/microbiología , Regulación de la Expresión Génica , Interacciones Huésped-Parásitos , Interacciones Huésped-Patógeno , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-12/genética , Interleucina-12/inmunología , Activación de Linfocitos , Ratones , Ratones Transgénicos , Células de Paneth/microbiología , Células de Paneth/parasitología , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunología , Células TH1/microbiología , Células TH1/parasitología , Toxoplasma/inmunología , Toxoplasmosis Animal/complicaciones , Toxoplasmosis Animal/inmunología , Toxoplasmosis Animal/parasitología , alfa-Defensinas/deficienciaRESUMEN
BACKGROUND AND AIMS: Microbial dysbiosis is associated with alcohol-related hepatitis (AH), with the mechanisms yet to be elucidated. The present study aimed to determine the effects of alcohol and zinc deficiency on Paneth cell (PC) antimicrobial peptides, α-defensins, and to define the link between PC dysfunction and AH. APPROACH AND RESULTS: Translocation of pathogen-associated molecular patterns (PAMPs) was determined in patients with severe AH and in a mouse model of alcoholic steatohepatitis. Microbial composition and PC function were examined in mice. The link between α-defensin dysfunction and AH was investigated in α-defensin-deficient mice. Synthetic human α-defensin 5 (HD5) was orally given to alcohol-fed mice to test the therapeutic potential. The role of zinc deficiency in α-defensin was evaluated in acute and chronic mouse models of zinc deprivation. Hepatic inflammation was associated with PAMP translocation and lipocalin-2 (LCN2) and chemokine (C-X-C motif) ligand 1 (CXCL1) elevation in patients with AH. Antibiotic treatment, lipopolysaccharide injection to mice, and in vitro experiments showed that PAMPs, but not alcohol, directly induced LCN2 and CXCL1. Chronic alcohol feeding caused systemic dysbiosis and PC α-defensin reduction in mice. Knockout of functional α-defensins synergistically affected alcohol-perturbed bacterial composition and the gut barrier and exaggerated PAMP translocation and liver damage. Administration of HD5 effectively altered cecal microbial composition, especially increased Akkermansia muciniphila, and reversed the alcohol-induced deleterious effects. Zinc-regulated PC homeostasis and α-defensins function at multiple levels, and dietary zinc deficiency exaggerated the deleterious effect of alcohol on PC bactericidal activity. CONCLUSIONS: Taken together, the study suggests that alcohol-induced PC α-defensin dysfunction is mediated by zinc deficiency and involved in the pathogenesis of AH. HD5 administration may represent a promising therapeutic approach for treating AH.
Asunto(s)
Traslocación Bacteriana , Hígado Graso Alcohólico/microbiología , Hígado Graso Alcohólico/fisiopatología , Microbiota/fisiología , Células de Paneth/fisiología , Zinc/deficiencia , alfa-Defensinas/deficiencia , Animales , Modelos Animales de Enfermedad , Disbiosis/etiología , Etanol/toxicidad , Hígado Graso Alcohólico/complicaciones , Humanos , Metaloproteinasa 7 de la Matriz/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microbiota/efectos de los fármacosRESUMEN
Crohn's disease may principally involve the whole gastrointestinal tract. Most commonly, the inflammation occurs in the small intestine and/or in the colon with stable disease location over the years. The pathogenesis of both disease phenotypes is complex, the likely primary defect lies in the innate rather than adaptive immunity, particularly in the chemical antimicrobial barrier of the mucosa. Crohn's ileitis is associated with a reduced expression of the Wnt signalling pathway transcription factor T-cell factor 4 (TCF4), which is regulating Paneth cell differentiation. As a result, the alpha-defensins and principal Paneth cell products HD5 and HD6 are deficiently expressed in ileal disease, independent of current inflammation. In contrast, Crohn's colitis is typically associated with an impaired induction of the beta-defensins HBD2 and HBD3 caused by fewer gene copy numbers in the gene locus of the beta-defensins on chromosome 8. This ileal and colonic defect in innate defence mediated by a deficiency of the protective alpha- and beta-defensins may enable the luminal microbes to invade the mucosa and trigger the inflammation. A better understanding of the exact molecular mechanisms behind ileal and colonic Crohn's disease may give rise to new therapeutic strategies based on a stimulation of the protective innate immune system.
Asunto(s)
Colon/inmunología , Enfermedad de Crohn/inmunología , Íleon/inmunología , Inmunidad Innata , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice , Colitis Ulcerosa/inmunología , Proteínas de Unión al ADN/metabolismo , Humanos , Inmunidad Mucosa , Células de Paneth/inmunología , Células Madre/inmunología , Factor de Transcripción 4 , Factores de Transcripción/metabolismo , alfa-Defensinas/deficiencia , beta-Defensinas/deficiencia , beta-Defensinas/genéticaRESUMEN
In about 70% of patients Crohn's disease (CD) affects the small intestine. This disease location is stable over time and associated with a genetic background different from isolated colonic disease. A characteristic feature of small intestinal host defense is the presence of Paneth cells at the bottom of the crypts of Lieberkühn. These cells produce different broad spectrum antimicrobial peptides (AMPs) most abundantly the α-defensins HD-5 and -6 (DEFA5 und DEFA6). In small intestinal Crohn's disease both these PC products are specifically reduced. As a functional consequence, ileal extracts from Crohn's disease patients are compromised in clearing bacteria and enteroadherent E. coli colonize the mucosa. Mechanisms for defective antimicrobial Paneth cell function are complex and include an association with a NOD2 loss of function mutation, a disturbance of the Wnt pathway transcription factor TCF7L2 (also known as TCF4), the autophagy factor ATG16L1, the endosomal stress protein XBP1, the toll-like receptor TLR9, the calcium mediated potassium channel KCNN4 as well as mutations or inactivation of HD5. Thus we conclude that small intestinal Crohn's disease is most likely a complex disease of the Paneth cell: Paneth's disease.
Asunto(s)
Enfermedad de Crohn/inmunología , Ileítis/inmunología , Células de Paneth/inmunología , Células de Paneth/patología , Animales , Proteínas Relacionadas con la Autofagia , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/inmunología , Proteínas Portadoras/genética , Proteínas Portadoras/inmunología , Enfermedad de Crohn/genética , Enfermedad de Crohn/fisiopatología , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/inmunología , Expresión Génica/inmunología , Humanos , Ileítis/genética , Ileítis/fisiopatología , Inmunidad Innata/genética , Inmunidad Innata/inmunología , Inmunidad Mucosa/genética , Inmunidad Mucosa/inmunología , Canales de Potasio de Conductancia Intermedia Activados por el Calcio/genética , Canales de Potasio de Conductancia Intermedia Activados por el Calcio/inmunología , Ratones , Mutación/inmunología , Proteína Adaptadora de Señalización NOD2/genética , Proteína Adaptadora de Señalización NOD2/inmunología , Células de Paneth/citología , Factores de Transcripción del Factor Regulador X , Transducción de Señal/genética , Transducción de Señal/inmunología , Receptor Toll-Like 9/genética , Receptor Toll-Like 9/inmunología , Factor de Transcripción 4 , Proteína 2 Similar al Factor de Transcripción 7/genética , Proteína 2 Similar al Factor de Transcripción 7/inmunología , Factores de Transcripción/genética , Factores de Transcripción/inmunología , Proteínas Wnt/genética , Proteínas Wnt/inmunología , Proteína 1 de Unión a la X-Box , alfa-Defensinas/deficiencia , alfa-Defensinas/genética , alfa-Defensinas/inmunologíaRESUMEN
Ileal Crohn's disease (CD), a chronic mucosal inflammation, is characterized by two pertinent features: a specific decrease of Paneth cell-produced antimicrobial alpha-defensins and the presence of mucosal-adherent bacteria. A mutation in NOD2, the muramyl dipeptide recognition receptor, is found in some patients, which leads to an even more pronounced alpha-defensin decrease. However, the underlying mechanism remains unclear for the majority of patients. In this study, we report a reduced expression in ileal CD of the Wnt-signaling pathway transcription factor Tcf-4, a known regulator of Paneth cell differentiation and alpha-defensin expression. Within specimens, the levels of Tcf-4 mRNA showed a high degree of correlation with both HD5 and HD6 mRNA. The levels of Tcf-4 mRNA were decreased in patients with ileal disease irrespective of degree of inflammation, but were not decreased in colonic CD or ulcerative colitis. As a functional indicator of Tcf-4 protein, quantitative binding analysis with nuclear extracts from small intestine biopsies to a Tcf-4 high-affinity binding site in the HD-5 and HD-6 promoters showed significantly reduced activity in ileal CD. Furthermore, a causal link was shown in a murine Tcf-4 knockout model, where the comparably reduced expression of Tcf-4 in heterozygous (+/-) mice was sufficient to cause a significant decrease of both Paneth cell alpha-defensin levels and bacterial killing activity. Finally, the association between Paneth cell alpha-defensins and Tcf-4 was found to be independent of the NOD2 genotype. This new link established between a human inflammatory bowel disease and the Wnt pathway/Tcf-4 provides a novel mechanism for pathogenesis in patients with ileal CD.
Asunto(s)
Enfermedad de Crohn/etiología , Ileítis/etiología , Células de Paneth/metabolismo , Factores de Transcripción TCF/metabolismo , Proteínas Wnt/metabolismo , alfa-Defensinas/deficiencia , Animales , Enfermedad de Crohn/genética , Enfermedad de Crohn/metabolismo , Humanos , Ileítis/genética , Ileítis/metabolismo , Ratones , Ratones Noqueados , Proteína Adaptadora de Señalización NOD2/genética , Regiones Promotoras Genéticas , Factores de Transcripción TCF/genética , Proteína 2 Similar al Factor de Transcripción 7 , Proteínas Wnt/genética , alfa-Defensinas/genéticaRESUMEN
Despite many years of intensive research the pathogenesis of inflammatory bowel diseases is still enigmatic. All efforts to identify the cause of Crohn's disease and ulcerative colitis in a dysregulation of specific immune mechanisms have failed. This review presents a novel pathogenetic concept, based on the expression of natural mucosal antibiotic peptides. These so called defensins are part of innate immunity and defend the mucosa as antimicrobial peptides against intraluminal potentially pathogenic and invasive bacteria. In contrast to ulcerative colitis, Crohn's disease is characterised by a diminished defensin expression. This defect may represent the molecular pathogenesis of this disease.
Asunto(s)
Colitis Ulcerosa/inmunología , Enfermedad de Crohn/inmunología , Traslocación Bacteriana/inmunología , Colitis Ulcerosa/diagnóstico , Enfermedad de Crohn/diagnóstico , Defensinas/deficiencia , Defensinas/fisiología , Humanos , Inmunidad Innata , Mucosa Intestinal/inmunología , alfa-Defensinas/deficiencia , alfa-Defensinas/fisiología , beta-Defensinas/deficiencia , beta-Defensinas/fisiologíaRESUMEN
BACKGROUND: The human platelet antigen (HPA) 1 through 5 and the human neutrophil antigen (HNA-1) systems are relevant to immune-related thrombocytopenia and neutropenia. The alloantigen distribution profiles in the population will aid in estimating the risk of alloimmunization. STUDY DESIGN AND METHODS: Genotyping of the genes that control the expression of the HPA-1 through -5 and HNA-1 systems in Taiwanese (n = 326) and Taiwan's indigenous peoples (n = 608) was performed by PCR with the sequence-specific primer (PCR-SSP) method. RESULTS: In the HPA system, HPA-1b and HPA-4b were absent among Taiwan's indigenous tribes and detected among other Taiwanese only with frequencies of <0.2 percent and <0.5 percent, respectively. The GP1BA*2 (HPA-2b) and GP1A*2 (HPA-5b) allele frequencies range from 1 percent to 7 percent and 0.4 percent to 3.5 percent among the two ethnic groups, respectively. GP2B*1 (HPA-3a) and GP2B*2 (HPA-3b) showed similar allele frequencies. In the HNA-1 system, the FCGR3B*1 (HNA-1a) allele frequency was about twice that of FCGR3B*2 (HNA-1b) in Taiwanese and also in most of the indigenous tribes. Three FCGR3B (HNA-1) null persons were found in one indigenous tribe (Ami tribe), for an FCGR3B null frequency of 19.8 percent. However, no FCGR3B*3 (HNA-1c) allele was detected in Taiwan. CONCLUSION: The frequencies of HPA-1b, -2b, and -5b in the Taiwanese population were much lower than those among whites. In Taiwan, all of the HNA-1 null found was due to the deletion of the FCGR3B gene, and this deletion may be widely distributed in the Ami tribe.