Interleukin 21 Drives a Hypermetabolic State and CD4+ T-Cell-Associated Pathogenicity in Chronic Intestinal Inflammation.

BACKGROUND & AIMS: Incapacitated regulatory T cells (Tregs) contribute to immune-mediated diseases. Inflammatory Tregs are evident during human inflammatory bowel disease; however, mechanisms driving the development of these cells and their function are not well understood. Therefore, we investigated the role of cellular metabolism in Tregs relevant to gut homeostasis. METHODS: Using human Tregs, we performed mitochondrial ultrastructural studies via electron microscopy and confocal imaging, biochemical and protein analyses using proximity ligation assay, immunoblotting, mass cytometry and fluorescence-activated cell sorting, metabolomics, gene expression analysis, and real-time metabolic profiling utilizing the Seahorse XF analyzer. We used a Crohn's disease single-cell RNA sequencing dataset to infer the therapeutic relevance of targeting metabolic pathways in inflammatory Tregs. We examined the superior functionality of genetically modified Tregs in CD4+ T-cell-induced murine colitis models. RESULTS: Mitochondria-endoplasmic reticulum appositions, known to mediate pyruvate entry into mitochondria via voltage-dependent anion channel 1 (VDAC1), are abundant in Tregs. VDAC1 inhibition perturbed pyruvate metabolism, eliciting sensitization to other inflammatory signals reversible by membrane-permeable methyl pyruvate supplementation. Notably, interleukin (IL) 21 diminished mitochondria-endoplasmic reticulum appositions, resulting in enhanced enzymatic function of glycogen synthase kinase 3 ß, a putative negative regulator of VDAC1, and a hypermetabolic state that amplified Treg inflammatory response. Methyl pyruvate and glycogen synthase kinase 3 ß pharmacologic inhibitor (LY2090314) reversed IL21-induced metabolic rewiring and inflammatory state. Moreover, IL21-induced metabolic genes in Tregs in vitro were enriched in human Crohn's disease intestinal Tregs. Adoptively transferred Il21r-/- Tregs efficiently rescued murine colitis in contrast to wild-type Tregs. CONCLUSIONS: IL21 triggers metabolic dysfunction associated with Treg inflammatory response. Inhibiting IL21-induced metabolism in Tregs may mitigate CD4+ T-cell-driven chronic intestinal inflammation.

CD8+ T cells recognizing a neuron-restricted antigen injure axons in a model of multiple sclerosis.

CD8+ T cells outnumber CD4+ cells in multiple sclerosis (MS) lesions associated with disease progression, but the pathogenic role and antigenic targets of these clonally expanded effectors are unknown. Based on evidence that demyelination is necessary but not sufficient for disease progression in MS, we previously hypothesized that CNS-infiltrating CD8+ T cells specific for neuronal antigens directly drive the axonal and neuronal injury that leads to cumulative neurologic disability in patients with MS. We now show that demyelination induced expression of MHC class I on neurons and axons and resulted in presentation of a neuron-specific neoantigen (synapsin promoter-driven chicken ovalbumin) to antigen-specific CD8+ T cells (anti-ovalbumin OT-I TCR-transgenic T cells). These neuroantigen-specific effectors surveilled the CNS in the absence of demyelination but were not retained. However, upon induction of demyelination via cuprizone intoxication, neuroantigen-specific CD8+ T cells proliferated, accumulated in the CNS, and damaged neoantigen-expressing neurons and axons. We further report elevated neuronal expression of MHC class I and ß2-microglobulin transcripts and protein in gray matter and white matter tracts in tissue from patients with MS. These findings support a pathogenic role for autoreactive anti-axonal and anti-neuronal CD8+ T cells in MS progression.

G9a Modulates Lipid Metabolism in CD4 T Cells to Regulate Intestinal Inflammation.

BACKGROUND & AIMS: Although T-cell intrinsic expression of G9a has been associated with murine intestinal inflammation, mechanistic insight into the role of this methyltransferase in human T-cell differentiation is ill defined, and manipulation of G9a function for therapeutic use against inflammatory disorders is unexplored. METHODS: Human naive T cells were isolated from peripheral blood and differentiated in vitro in the presence of a G9a inhibitor (UNC0642) before being characterized via the transcriptome (RNA sequencing), chromatin accessibility (assay for transposase-accessible chromatin by sequencing), protein expression (cytometry by time of flight, flow cytometry), metabolism (mitochondrial stress test, ultrahigh performance liquid chromatography-tandem mas spectroscopy) and function (T-cell suppression assay). The in vivo role of G9a was assessed using 3 murine models. RESULTS: We discovered that pharmacologic inhibition of G9a enzymatic function in human CD4 T cells led to spontaneous generation of FOXP3+ T cells (G9a-inibitors-T regulatory cells [Tregs]) in vitro that faithfully reproduce human Tregs, functionally and phenotypically. Mechanistically, G9a inhibition altered the transcriptional regulation of genes involved in lipid biosynthesis in T cells, resulting in increased intracellular cholesterol. Metabolomic profiling of G9a-inibitors-Tregs confirmed elevated lipid pathways that support Treg development through oxidative phosphorylation and enhanced lipid membrane composition. Pharmacologic G9a inhibition promoted Treg expansion in vivo upon antigen (gliadin) stimulation and ameliorated acute trinitrobenzene sulfonic acid-induced colitis secondary to tissue-specific Treg development. Finally, Tregs lacking G9a expression (G9a-knockout Tregs) remain functional chronically and can rescue T-cell transfer-induced colitis. CONCLUSION: G9a inhibition promotes cholesterol metabolism in T cells, favoring a metabolic profile that facilitates Treg development in vitro and in vivo. Our data support the potential use of G9a inhibitors in the treatment of immune-mediated conditions including inflammatory bowel disease.

Fracture characteristics of anterior resin-bonded zirconia-fixed partial dentures.

Resin-bonded fixed partial dentures (RBFPD) are used as a minimal invasive, tooth-preventing alternative for replacing anterior teeth. Zirconia cantilever restorations were supposed to show sufficient strength for a clinical application. The aim of this investigation was to determine the fracture characteristics of cantilever and two-retainer RBFPD, which are fabricated by computer-manufactured high-strength zirconia. Human incisors and canines were used to form three groups of 14 RBFPDs with different types of preparation: group 1, an invasive cantilever; group 2, a minimal-invasive cantilever and group 3, a two-retainer RBFPD control. After thermal cycling and mechanical loading, which was performed to simulate oral service, all restorations were loaded to fracture in a universal testing machine. One half of the specimens were investigated as a control without simulated service. Mode of failure was determined for the three designs. Both cantilever groups showed comparable fracture resistance of 227 N (no. 1) and 210 N (no. 2) before thermal cycling and mechanical loading. The resistance after aging was reduced to 210 N for the invasive cantilever RBFPD and to 179 N for the minimal invasive group. Three-unit RBFPDs showed a significantly higher (p < 0.02) fracture resistance than cantilever bridges before (426 N) as well as after aging (360 N). Predominant failure was FPD and retainer fracture for the invasive cantilever design, debonding for the minimal cantilever design and RBFPD fracture for the two-retainer design. The present study revealed a significantly higher fracture resistance for two-retainer RBFPDs than for cantilever RBFPDs. The frequency of adhesive debonding increased for non-retentive prepared cantilever RBFPDs.

Cidade saudável: uma experiência de interdisciplinaridade e intersetorialidade / Heathy city: an interdisciplinary and intersectorial experience

Estratégias de promoção da saúde, como projetos de cidades saudáveis, vêm sendo desenvolvidas por alguns municípios brasileiros, com o intuito de responder às mudanças decorrentes da crescente urbanização e de suas consequências para a saúde e qualidade de vida das populações. Dada a complexidade dos problemas enfrentados, pressupostos como a interdisciplinaridade, a intersetorialidade e a participação social são considerados fundamentos que devem orientar uma nova prática, buscando superar uma lógica de gestão municipal predominante: verticalizada, setorial e dicotômica. O aparato governamental de algumas cidades envolvidas no movimento por cidades saudáveis tem experimentado diferentes formas de organização para dar respostas articuladas e integradas aos problemas e desafios presentes.

Políticas públicas e investimentos: a intersetorialidade / Public police and investment: the intersectoriality

Busca, dentro dos contornos de uma concepçäo específica de Estado, no processo de descentralizaçäo administrativa brasileira, as responsabilidades e o papel dos governos locais na definiçäo de políticas públicas