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Gut bacteria are linked to neurodegenerative diseases but the risk factors beyond microbiota composition are limited. Here we used a pre-clinical model of multiple sclerosis (MS), experimental autoimmune encephalomyelitis (EAE), to identify microbial risk factors. Mice with different genotypes and complex microbiotas or six combinations of a synthetic human microbiota were analysed, resulting in varying probabilities of severe neuroinflammation. However, the presence or relative abundances of suspected microbial risk factors failed to predict disease severity. Akkermansia muciniphila, often associated with MS, exhibited variable associations with EAE severity depending on the background microbiota. Significant inter-individual disease course variations were observed among mice harbouring the same microbiota. Evaluation of microbial functional characteristics and host immune responses demonstrated that the immunoglobulin A coating index of certain bacteria before disease onset is a robust individualized predictor of disease development. Our study highlights the need to consider microbial community networks and host-specific bidirectional interactions when aiming to predict severity of neuroinflammation.
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BACKGROUND: The gut microbiota plays an important role in the development of behavior and immunity in infants and juveniles. Early weaning (EW), a form of social stress in mice, leads to increased anxiety and an enhanced stress response in the hypothalamic-pituitary-adrenal axis during adulthood. Early life stress also modulates the immune system and increases vulnerability to infection. However, studies investigating the causal relationships among juvenile stress, microbiota changes, and immune and behavioral deficits are limited. Therefore, we hypothesized that EW alters gut microbiota composition and impairs the development of the nervous and immune systems. RESULTS: EW mice moved longer distances in the marble-burying test and had longer immobility times in the tail suspension test than normal weaning (NW) mice. In parallel, the gut microbiome composition differed between NW and EW mice, and the abundance of Erysipelotrichacea in EW mice at 8 weeks of age was lower than that in NW mice. In an empirical study, germ-free mice colonized with the gut microbiota of EW mice (GF-EW mice) demonstrated higher depressive behavior than GF mice colonized with normal weaning microbiota (GF-NW mice). Immune cell profiles were also affected by the EW microbiota colonization; the number of CD4 + T cells in the spleen was reduced in GF-EW mice. CONCLUSION: Our results suggest that EW-induced alterations in the gut microbiota cause depressive behaviors and modulate the immune system.
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Mammalian gut microbes colonize the intestinal tract of their host and adapt to establish a microbial ecosystem. The host diet changes the nutrient profile of the intestine and has a high impact on microbiota composition. Genetic mutations in Escherichia coli, a prevalent species in the human gut, allow for adaptation to the mammalian intestine, as reported in previous studies. However, the extent of colonization fitness in the intestine elevated by genetic mutation and the effects of diet change on these mutations in E. coli are still poorly known. Here, we show that notable mutations in sugar metabolism-related genes (gatC, araC, and malI) were detected in the E. coli K-12 genome just 2 weeks after colonization in the germ-free mouse intestine. In addition to elevated fitness by deletion of gatC, as previously reported, deletion of araC and malI also elevated E. coli fitness in the murine intestine in a host diet-dependent manner. In vitro cultures of medium containing nutrients abundant in the intestine (e.g., galactose, N-acetylglucosamine, and asparagine) also showed increased E. coli fitness after deletion of the genes-of-interest associated with their metabolism. Furthermore, the host diet was found to influence the developmental trajectory of gene mutations in E. coli. Taken together, we suggest that genetic mutations in E. coli are selected in response to the intestinal environment, which facilitates efficient utilization of nutrients abundant in the intestine under laboratory conditions. Our study offers some insight into the possible adaptation mechanisms of gut microbes.IMPORTANCEThe gut microbiota is closely associated with human health and is greatly impacted by the host diet. Bacteria such as Escherichia coli live in the gut all throughout the life of a human host and adapt to the intestinal environment. Adaptive mutations in E. coli are reported to enhance fitness in the mammalian intestine, but to what extent is still poorly known. It is also unknown whether the host diet affects what genes are mutated and to what extent fitness is affected. This study suggests that genetic mutations in the E. coli K-12 strain are selected in response to the intestinal environment and facilitate efficient utilization of abundant nutrients in the germ-free mouse intestine. Our study provides a better understanding of these intestinal adaptation mechanisms of gut microbes.
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Ecossistema , Escherichia coli , Humanos , Animais , Camundongos , Escherichia coli/genética , Dieta , Intestinos/microbiologia , Mutação , MamíferosRESUMO
Multiple stenotic lesions may restrict the access sites for endovascular therapy in the lower extremity arteries. Because guide sheaths used for endovascular therapy have recently become easier to insert, they are directly inserted into the posterior tibial or dorsalis pedis artery to perform the transtibial approach. We herein describe an 81-year-old man who was admitted to our hospital because of claudication of the left lower extremity. He had a history of left iliofemoral and femorofemoral bypass surgery. The patient's symptom was due to a stenotic lesion extending from the left common femoral artery to the distal part of the left superficial femoral artery. In an angiographic procedure using the antegrade approach via the right radial artery, a multipurpose catheter became stuck in the middle of the left iliofemoral bypass. The antegrade ipsilateral approach was too close to the stenotic lesion for the insertion of the guide sheath. Therefore, a retrograde approach using a 5-French guide sheath inserted via the dorsalis pedis artery was successfully performed.
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The prevalence of peripheral artery disease (PAD) has been increasing in parallel with the increasing prevalence of the atherosclerotic disease. Therefore, we have to be familiar with the diagnostic approach used for ischemic symptoms in the lower limbs. Adventitial cystic disease (ACD) is rare but not negligible as one of the differential diagnoses of intermittent claudication (IC). Although duplex ultrasound and magnetic resonance imaging (MRI) are helpful tools for the diagnosis of ACD, further imaging modality is needed to avoid misdiagnosis. A 64-year-old man with a mitral valve prosthesis presented to our hospital with a one-month history of IC in the right calf after walking for approximately 50 meters. On physical examination, the pulse in the right popliteal artery was not palpable, nor were the dorsal pedis artery and posterior tibial artery, although there were no other symptoms of ischemia. His right ankle-brachial index (ABI) was 1.12 at rest but decreased to 0.50 after exercise. Three-dimensional computed tomography (CT) angiography revealed a severe stenotic lesion approximately 70 mm long in the right popliteal artery. Therefore, we diagnosed PAD in the right lower limb and planned endovascular therapy. The stenotic lesion was markedly reduced on catheter angiography when compared with CT angiography. However, intravascular ultrasound (IVUS) detected little atherosclerosis and cystic lesions within the wall in the right popliteal artery that did not involve the arterial lumen. Especially, IVUS clearly demonstrated that the crescent-shaped cyst compressed the arterial lumen eccentrically and other cysts surrounded the lumen circumferentially like petals. Because IVUS revealed these cysts to be extravascular structures, the patient was subsequently thought to have ACD of the right popliteal artery. Fortunately, his cysts reduced in size spontaneously and his symptoms disappeared. We have monitored the patient's symptoms, ABI, and findings on duplex ultrasound for seven years, during which there has been no recurrence. In this case, we diagnosed ACD in the popliteal artery by IVUS rather than duplex ultrasound and MRI.
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Although recent studies have highlighted the impact of gut microbes on the progression of obesity and its comorbidities, it is not fully understood how these microbes promote these disorders, especially in terms of the role of microbial metabolites. Here, we report that Fusimonas intestini, a commensal species of the family Lachnospiraceae, is highly colonized in both humans and mice with obesity and hyperglycemia, produces long-chain fatty acids such as elaidate, and consequently facilitates diet-induced obesity. High fat intake altered the expression of microbial genes involved in lipid production, such as the fatty acid metabolism regulator fadR. Monocolonization with a FadR-overexpressing Escherichia coli exacerbated the metabolic phenotypes, suggesting that the change in bacterial lipid metabolism is causally involved in disease progression. Mechanistically, the microbe-derived fatty acids impaired intestinal epithelial integrity to promote metabolic endotoxemia. Our study thus provides a mechanistic linkage between gut commensals and obesity through the overproduction of microbe-derived lipids.
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Ácidos Graxos , Microbioma Gastrointestinal , Humanos , Animais , Camundongos , Dieta Hiperlipídica , Obesidade/metabolismo , Bactérias/genética , Camundongos Endogâmicos C57BLRESUMO
BACKGROUND: ST-elevation is one of the most valuable electrocardiogram findings to diagnose acute myocardial infarction. However, more than a quarter of acute coronary occlusions are missed by this criterion, causing a delay in revascularization. Therefore, there should be awareness of the limitations of the current criteria and new electrocardiographic findings are required as a diagnostic tool to compensate for them. The Aslanger pattern is a specific electrocardiographic finding in acute inferior myocardial infarction with multivessel disease and allows the detection of inferior myocardial infarction that does not show ST-elevation, leading to rapid revascularization. However, in patients with the Aslanger pattern, the hemodynamic characteristics, such as the rate of shock and the use of mechanical circulatory support, as well as prognostic characteristics such as the in-hospital mortality rate, have not yet been clarified. METHODS: In this study, we retrospectively surveyed the current practice on the basis of ST-elevation myocardial infarction (STEMI) criteria in patients with acute coronary artery occlusion presenting with inferior myocardial infarction. We examined the clinical characteristics of the Aslanger pattern. RESULTS: Based on the STEMI criteria, 71.8% (51/72) of patients were diagnosed with STEMI from an acute electrocardiogram, and 28.2% (21/78) were diagnosed with non-STEMI. As expected, ruling out in all acute coronary artery occlusions using STEMI criteria alone was difficult. A total of 48% of patients with non-STEMI had the Aslanger pattern. In addition, 80% of patients with the Aslanger pattern had multivessel disease, 30% had the use of the mechanical circulatory support, and 20% had in-hospital mortality. CONCLUSION: This study suggests that the Aslanger pattern is useful not only for diagnosis, but also for predicting hemodynamic collapse and a poor prognosis. Therefore, we should share information on Aslanger pattern with other physicians and use this pattern in daily practice.
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The prevalence of autoimmune diseases (ADs) worldwide has rapidly increased over the past few decades. Thus, in addition to the classical risk factors for ADs, such as genetic polymorphisms, infections and smoking, environmental triggers have been considered. Recent sequencing-based approaches have revealed that patients with extra-intestinal ADs, such as multiple sclerosis, rheumatoid arthritis, type 1 diabetes and systemic lupus erythematosus, have distinct gut microbiota compositions compared to healthy controls. Faecal microbiota transplantation or inoculation with specific microbes in animal models of ADs support the hypothesis that alterations of gut microbiota influence autoimmune responses and disease outcome. Here, we describe the compositional and functional changes in the gut microbiota in patients with extra-intestinal AD and discuss how the gut microbiota affects immunity. Moreover, we examine how the gut microbiota might be modulated in patients with ADs as a potential preventive or therapeutic approach.
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Doenças Autoimunes , Diabetes Mellitus Tipo 1 , Microbioma Gastrointestinal , Enteropatias , Lúpus Eritematoso Sistêmico , Animais , Humanos , Lúpus Eritematoso Sistêmico/etiologia , Lúpus Eritematoso Sistêmico/terapia , Fatores de Risco , DisbioseRESUMO
Entrapment of devices, such as a Rota bar, an extension catheter, or an intravascular ultrasound device, during percutaneous coronary intervention has been reported and bailout strategies have been discussed. However, there have been few reports on entrapment of devices during endovascular treatment (EVT). A 70-year-old man was referred to our clinic for the management of rest pain in his left lower limb. His left ankle-brachial index was unmeasurable and computed tomography angiography revealed total occlusion of the left common, external iliac, and superficial femoral arteries (SFA). He was diagnosed as having symptomatic limb-threatening ischemia and EVT was planned. The first EVT was performed on an occluding lesion in the left iliac artery. We used a transradial approach and deployed two bare nitinol stents in the left iliac artery without complications. One week after the first EVT, the second EVT was performed on an occluding lesion in the left SFA. A 6.0-French (Fr) guide sheath was inserted antegradely through the left common femoral artery. The occluded lesion was dilated with a 4.0 mm plain balloon, following which intravascular ultrasound revealed a localized severe stenotic lesion in the distal part of the SFA. A 6.0 mm drug-eluting stent was deployed to cover the stenotic lesion in the distal part of the SFA without pre-dilation; however, the stenotic lesion did not dilate sufficiently. When we attempted to extract the stent delivery catheter, we could not detach its tip from the localized severe stenotic lesion and were unable to remove it by force or external compression. Therefore, we decided to implement a double guide technique by inserting a 4.0-Fr sheath simultaneously into the left common femoral artery adjacent to the first puncture site together with another 0.014-inch guidewire via a 4.0-Fr sheath to get past the lesion in which the catheter tip was embedded. We then used a 3.0-mm plain balloon to dilate the severe stenotic lesion sufficiently to enable the removal of the stent delivery catheter. Another 6.0-mm drug-eluting stent was then deployed, after the first stent, to cover the occluded lesion in the middle part of the SFA. Hemostasis was safely achieved at both puncture sites by manual compression. A double guide technique, as in percutaneous coronary intervention, is useful for the bailout of an entrapped device during EVT. Careful consideration of the access site and size and length of the second guide sheath are necessary.
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Background: The effect of oral microbiota on the intestinal microbiota has garnered growing attention as a mechanism linking periodontal diseases to systemic diseases. However, the salivary microbiota is diverse and comprises numerous bacteria with a largely similar composition in healthy individuals and periodontitis patients. Aim: We explored how health-associated and periodontitis-associated salivary microbiota differently colonized the intestine and their subsequent systemic effects. Methods: The salivary microbiota was collected from a healthy individual and a periodontitis patient and gavaged into C57BL/6NJcl[GF] mice. Gut microbial communities, hepatic gene expression profiles, and serum metabolites were analyzed. Results: The gut microbial composition was significantly different between periodontitis-associated microbiota-administered (PAO) and health-associated oral microbiota-administered (HAO) mice. The hepatic gene expression profile demonstrated a distinct pattern between the two groups, with higher expression of lipid and glucose metabolism-related genes. Disease-associated metabolites such as 2-hydroxyisobutyric acid and hydroxybenzoic acid were elevated in PAO mice. These metabolites were significantly correlated with characteristic gut microbial taxa in PAO mice. Conversely, health-associated oral microbiota were associated with higher levels of beneficial serum metabolites in HAO mice. Conclusion: The multi-omics approach used in this study revealed that periodontitis-associated oral microbiota is associated with the induction of disease phenotype when they colonized the gut of germ-free mice.
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The bacterial microbiota works as a community that consists of many individual organisms, i.e., cells. To fully understand the function of bacterial microbiota, individual cells must be identified; however, it is difficult with current techniques. Here, we develop a method, Barcoding Bacteria for Identification and Quantification (BarBIQ), which classifies single bacterial cells into taxa-named herein cell-based operational taxonomy units (cOTUs)-based on cellularly barcoded 16S rRNA sequences with single-base accuracy, and quantifies the cell number for each cOTU in the microbiota in a high-throughput manner. We apply BarBIQ to murine cecal microbiotas and quantify in total 3.4 × 105 bacterial cells containing 810 cOTUs. Interestingly, we find location-dependent global differences in the cecal microbiota depending on the dietary vitamin A deficiency, and more differentially abundant cOTUs at the proximal location than the distal location. Importantly, these location differences are not clearly shown by conventional 16S rRNA gene-amplicon sequencing methods, which quantify the 16S rRNA genes, not the cells. Thus, BarBIQ enables microbiota characterization with the identification and quantification of individual constituent bacteria, which is a cornerstone for microbiota studies.
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Sequenciamento de Nucleotídeos em Larga Escala , Microbiota , Animais , Bactérias/genética , DNA Bacteriano/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Camundongos , Microbiota/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
The bacterial composition of the gut lumen and mucosa is distinct and the mucosa-associated bacteria are thought to play a more critical role in interactions with the host immune system. However, limited studies of the gut mucosal microbiota in humans have been available due to methodological challenges. Here, we evaluated the potential use of colonic lavage samples for mucosal microbiota analysis in humans. Among the different types of colonic mucosal samples collected from healthy volunteers, the lavage samples contained a higher amount of bacterial DNA and were less contaminated with host DNA compared to mucosal brushing (brush) and biopsy. Although 16S gene amplicon sequencing showed that the bacterial composition of the lavage was intermediate between that of feces and biopsy, mucosal bacteria abundant in the biopsy were also enriched in lavage samples. Furthermore, differences in mucosal microbes between non-smokers and smokers were detectable in lavage samples. Our data emphasize that colonic lavage is suitable for analysis of the mucosal microbiota. Given its minimal invasiveness and high bacterial DNA content, the colonic lavage will promote research on the human mucosal microbiota, especially in gastrointestinal disorders.
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Colo/microbiologia , Endoscopia , Microbioma Gastrointestinal/genética , Bactérias/genética , Bactérias/isolamento & purificação , Fumar Cigarros , DNA Bacteriano , Fezes/microbiologia , Humanos , Mucosa Intestinal/microbiologia , Metagenômica/métodos , RNA Ribossômico 16S/genéticaRESUMO
Background & Aims: Periodontitis increases the risk of nonalcoholic fatty liver disease (NAFLD); however, the underlying mechanisms are unclear. Here, we show that gut dysbiosis induced by oral administration of Porphyromonas gingivalis, a representative periodontopathic bacterium, is involved in the aggravation of NAFLD pathology. Methods: C57BL/6N mice were administered either vehicle, P. gingivalis, or Prevotella intermedia, another periodontopathic bacterium with weaker periodontal pathogenicity, followed by feeding on a choline-deficient, l-amino acid-defined, high-fat diet with 60 kcal% fat and 0.1% methionine (CDAHFD60). The gut microbial communities were analyzed by pyrosequencing the 16S ribosomal RNA genes. Metagenomic analysis was used to determine the relative abundance of the Kyoto Encyclopedia of Genes and Genomes pathways encoded in the gut microbiota. Serum metabolites were analyzed using nuclear magnetic resonance-based metabolomics coupled with multivariate statistical analyses. Hepatic gene expression profiles were analyzed via DNA microarray and quantitative polymerase chain reaction. Results: CDAHFD60 feeding induced hepatic steatosis, and in combination with bacterial administration, it further aggravated NAFLD pathology, thereby increasing fibrosis. Gene expression analysis of liver samples revealed that genes involved in NAFLD pathology were perturbed, and the two bacteria induced distinct expression profiles. This might be due to quantitative and qualitative differences in the influx of bacterial products in the gut because the serum endotoxin levels, compositions of the gut microbiota, and serum metabolite profiles induced by the ingested P. intermedia and P. gingivalis were different. Conclusions: Swallowed periodontopathic bacteria aggravate NAFLD pathology, likely due to dysregulation of gene expression by inducing gut dysbiosis and subsequent influx of gut bacteria and/or bacterial products.
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Microbioma Gastrointestinal , Hepatopatia Gordurosa não Alcoólica/microbiologia , Porphyromonas gingivalis , Prevotella intermedia , Administração Oral , Animais , Deficiência de Colina , Dieta Hiperlipídica , Fezes/microbiologia , Células Hep G2 , Humanos , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/patologia , RNA Ribossômico 16SRESUMO
The balance between bacterial colonization and its containment in the intestine is indispensable for the symbiotic relationship between humans and their bacteria. One component to maintain homeostasis at the mucosal surfaces is immunoglobulin A (IgA), the most abundant immunoglobulin in mammals1,2. Several studies have revealed important characteristics of poly-reactive IgA3,4, which is produced naturally without commensal bacteria. Considering the dynamic changes within the gut environment, however, it remains uncertain how the commensal-reactive IgA pool is shaped and how such IgA affects the microbial community. Here we show that acetate-one of the major gut microbial metabolites-not only increases the production of IgA in the colon, but also alters the capacity of the IgA pool to bind to specific microorganisms including Enterobacterales. Induction of commensal-reactive IgA and changes in the IgA repertoire by acetate were observed in mice monocolonized with Escherichia coli, which belongs to Enterobacterales, but not with the major commensal Bacteroides thetaiotaomicron, which suggests that acetate directs selective IgA binding to certain microorganisms. Mechanistically, acetate orchestrated the interactions between epithelial and immune cells, induced microbially stimulated CD4 T cells to support T-cell-dependent IgA production and, as a consequence, altered the localization of these bacteria within the colon. Collectively, we identified a role for gut microbial metabolites in the regulation of differential IgA production to maintain mucosal homeostasis.
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Acetatos/farmacologia , Bactérias/imunologia , Microbioma Gastrointestinal/imunologia , Imunoglobulina A/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Colo/imunologia , Dieta , Ácidos Graxos Voláteis/metabolismo , Homeostase/imunologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , SimbioseRESUMO
Accumulating evidence indicates that gut microorganisms have a pathogenic role in autoimmune diseases, including in multiple sclerosis1. Studies of experimental autoimmune encephalomyelitis (an animal model of multiple sclerosis)2,3, as well as human studies4-6, have implicated gut microorganisms in the development or severity of multiple sclerosis. However, it remains unclear how gut microorganisms act on the inflammation of extra-intestinal tissues such as the spinal cord. Here we show that two distinct signals from gut microorganisms coordinately activate autoreactive T cells in the small intestine that respond specifically to myelin oligodendrocyte glycoprotein (MOG). After induction of experimental autoimmune encephalomyelitis in mice, MOG-specific CD4+ T cells are observed in the small intestine. Experiments using germ-free mice that were monocolonized with microorganisms from the small intestine demonstrated that a newly isolated strain in the family Erysipelotrichaceae acts similarly to an adjuvant to enhance the responses of T helper 17 cells. Shotgun sequencing of the contents of the small intestine revealed a strain of Lactobacillus reuteri that possesses peptides that potentially mimic MOG. Mice that were co-colonized with these two strains showed experimental autoimmune encephalomyelitis symptoms that were more severe than those of germ-free or monocolonized mice. These data suggest that the synergistic effects that result from the presence of these microorganisms should be considered in the pathogenicity of multiple sclerosis, and that further study of these microorganisms may lead to preventive strategies for this disease.
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Encefalomielite Autoimune Experimental/microbiologia , Microbioma Gastrointestinal/imunologia , Inflamação/patologia , Medula Espinal/patologia , Linfócitos T/imunologia , Linfócitos T/patologia , Animais , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/patologia , Encefalomielite Autoimune Experimental/prevenção & controle , Feminino , Vida Livre de Germes , Inflamação/imunologia , Intestino Delgado/imunologia , Intestino Delgado/microbiologia , Intestino Delgado/patologia , Limosilactobacillus reuteri/química , Limosilactobacillus reuteri/imunologia , Limosilactobacillus reuteri/patogenicidade , Masculino , Camundongos , Esclerose Múltipla/imunologia , Esclerose Múltipla/microbiologia , Esclerose Múltipla/patologia , Glicoproteína Mielina-Oligodendrócito/química , Glicoproteína Mielina-Oligodendrócito/imunologia , Medula Espinal/imunologia , Células Th17/imunologia , Células Th17/patologiaRESUMO
Acquired immune responses are initiated by activation of CD4+ helper T (Th) cells via recognition of antigens presented by conventional dendritic cells (cDCs). DCs instruct Th-cell polarization program into specific effector Th subset, which will dictate the type of immune responses. Hence, it is important to unravel how differentiation and/or activation of DC are linked with Th-cell-intrinsic mechanism that directs differentiation toward a specific effector Th subset. Here, we show that loss of Runx/Cbfß transcription factors complexes during DC development leads to loss of CD103+CD11b+ cDC2s and alters characteristics of CD103-CD11b+ cDCs in the intestine, which was accompanied with impaired differentiation of Rorγt+ Th17 cells and type 3 Rorγt+ regulatory T cells. We also show that a Runx-binding enhancer in the Rorc gene is essential for T cells to integrate cDC-derived signals to induce Rorγt expression. These findings reveal that Runx/Cbfß complexes play crucial and complementary roles in cDCs and Th cells to shape converging type 3 immune responses.
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Subunidade alfa 2 de Fator de Ligação ao Core/metabolismo , Subunidade alfa 3 de Fator de Ligação ao Core/metabolismo , Subunidade beta de Fator de Ligação ao Core/metabolismo , Células Dendríticas/metabolismo , Mucosa Intestinal/citologia , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Linfócitos T Reguladores/metabolismo , Células Th17/metabolismo , Imunidade Adaptativa , Animais , Diferenciação Celular/imunologia , Células Cultivadas , Subunidade alfa 2 de Fator de Ligação ao Core/genética , Subunidade alfa 3 de Fator de Ligação ao Core/genética , Subunidade beta de Fator de Ligação ao Core/genética , Células Dendríticas/imunologia , Mucosa Intestinal/imunologia , Camundongos , Camundongos Transgênicos , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Linfócitos T Reguladores/imunologia , Células Th17/imunologiaRESUMO
Interleukin-15 (IL-15) is a cytokine critical for maintenance of intestinal intra-epithelial lymphocytes (IELs), especially CD8ααâ+ IELs (CD8αα IELs). In the intestine, IL-15 is produced by intestinal epithelial cells (IECs), blood vascular endothelial cells (BECs) and hematopoietic cells. However, the precise role of intestinal IL-15 on IELs is still unknown. To address the question, we generated two kinds of IL-15 conditional knockout (IL-15cKO) mice: villin-Cre (Vil-Cre) and Tie2-Cre IL-15cKO mice. IEC-derived IL-15 was specifically deleted in Vil-Cre IL-15cKO mice, whereas IL-15 produced by BECs and hematopoietic cells was deleted in Tie2-Cre IL-15cKO mice. The cell number and frequency of CD8αα IELs and NK IELs were significantly reduced in Vil-Cre IL-15cKO mice. By contrast, CD8αα IELs were unchanged in Tie2-Cre IL-15cKO mice, indicating that IL-15 produced by BECs and hematopoietic cells is dispensable for CD8αα IELs. Expression of an anti-apoptotic factor, Bcl-2, was decreased, whereas Fas expression was increased in CD8αα IELs of Vil-Cre IL-15cKO mice. Forced expression of Bcl-2 by a Bcl-2 transgene partially restored CD8αα IELs in Vil-Cre IL-15cKO mice, suggesting that some IL-15 signal other than Bcl-2 is required for maintenance of CD8αα IELs. Furthermore, granzyme B production was reduced, whereas PD-1 expression was increased in CD8αα IELs of Vil-Cre IL-15cKO mice. These results collectively suggested that IEC-derived IL-15 is essential for homeostasis of IELs by promoting their survival and functional maturation.
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Células Endoteliais/imunologia , Interleucina-15/imunologia , Intestinos/citologia , Intestinos/imunologia , Linfócitos Intraepiteliais/citologia , Linfócitos Intraepiteliais/imunologia , Animais , Feminino , Interleucina-15/deficiência , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos TransgênicosRESUMO
BACKGROUND: More than 2500 variants of the low-density lipoprotein receptor (LDLR) gene have been reported in familial hypercholesterolemia (FH). However, the effects of these variants on the pathophysiology of FH have not been fully clarified. OBJECTIVE: Our aim was to examine whether the c.2579C>T (p.A860V) variant of the LDLR gene affects the phenotype of FH. We present 2 index cases harboring biallelic LDLR variants, including the c.2579C>T (p.A860V) variant, which is defined as having uncertain significance in ClinVar. METHODS: Genetic analysis was performed for coding regions of the LDLR and proprotein convertase subtilisin/kexin type 9 (PCSK9) genes in 2 families. Detailed clinical and biochemical data were gathered from family members. RESULTS: In one family, the index case involved a patient who harbored biallelic A860V and c.1528-1529insA (p.T510Nfs) LDLR variants and had 8 children; the affected children had the p.T510Nfs variant, and the unaffected children had the A860V variant. In another family, the patient involved in the index case and his sister had biallelic A860V and c.1845+2T>C LDLR variants. There was no difference in FH phenotype between these siblings and their relatives who were heterozygous for the c.1845+2T>C variant. In addition, the allele frequency of the A860V variant (0.0062/0.0095) in the Japanese population, as indicated by 2 databases, was higher than expected based on the prevalence of heterozygous FH in the Japanese population (0.002-0.005). CONCLUSIONS: This is the first report to show using pedigree-based genetic analysis that the A860V variant of the LDLR gene is a benign variant.
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Hiperlipoproteinemia Tipo II/genética , Linhagem , Polimorfismo de Nucleotídeo Único , Receptores de LDL/genética , Adolescente , Criança , Feminino , Frequência do Gene , Humanos , Hiperlipoproteinemia Tipo II/patologia , Masculino , Pessoa de Meia-Idade , FenótipoRESUMO
Accumulating evidence suggests that dysbiosis plays a role in the pathogenesis of intestinal diseases including inflammatory bowel disease (IBD) as well as extra-intestinal disorders. As a modulator of the intestinal microbiota, we isolated a mouse monoclonal IgA antibody (clone W27) with high affinities for multiple commensal bacteria, but not for beneficial bacteria such as Lactobacillus casei (L. casei). Via specific recognition of an epitope in serine hydroxymethyltransferase (SHMT), a bacterial metabolic enzyme, W27 IgA selectively inhibited the in vitro growth of bound bacteria, including Escherichia coli (E. coli), while having no effect on unbound beneficial bacteria such as L. casei. By modulating the gut microbiota in vivo, oral administration of W27 IgA effectively prevented development of colitis in several mouse models. Here we discuss how intestinal IgA modulates the gut microbiota through recognition of SHMT.
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Anticorpos Antibacterianos/administração & dosagem , Microbioma Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/microbiologia , Imunoglobulina A/administração & dosagem , Fatores Imunológicos/administração & dosagem , Administração Oral , Animais , Antibacterianos/administração & dosagem , Anticorpos Monoclonais/administração & dosagem , Glicina Hidroximetiltransferase/imunologia , Inibidores do Crescimento/administração & dosagem , CamundongosRESUMO
Immunoglobulin A (IgA) is the main antibody isotype secreted into the intestinal lumen. IgA plays a critical role in the defence against pathogens and in the maintenance of intestinal homeostasis. However, how secreted IgA regulates gut microbiota is not completely understood. In this study, we isolated monoclonal IgA antibodies from the small intestine of healthy mouse. As a candidate for an efficient gut microbiota modulator, we selected a W27 IgA, which binds to multiple bacteria, but not beneficial ones such as Lactobacillus casei. W27 could suppress the cell growth of Escherichia coli but not L. casei in vitro, indicating an ability to improve the intestinal environment. Indeed W27 oral treatment could modulate gut microbiota composition and have a therapeutic effect on both lymphoproliferative disease and colitis models in mice. Thus, W27 IgA oral treatment is a potential remedy for inflammatory bowel disease, acting through restoration of host-microbial symbiosis.