RESUMO
Taste bud cells regenerate throughout life. Taste bud maintenance depends on continuous replacement of senescent taste cells with new ones generated by adult taste stem cells. More than a century ago it was shown that taste buds degenerate after their innervating nerves are transected and that they are not restored until after reinnervation by distant gustatory ganglion neurons. Thus, neuronal input, likely via neuron-supplied factors, is required for generation of differentiated taste cells and taste bud maintenance. However, the identity of such a neuron-supplied niche factor(s) remains unclear. Here, by mining a published RNA-sequencing dataset of geniculate ganglion neurons and by in situ hybridization, we demonstrate that R-spondin-2, the ligand of Lgr5 and its homologs Lgr4/6 and stem-cell-expressed E3 ligases Rnf43/Znrf3, is expressed in nodose-petrosal and geniculate ganglion neurons. Using the glossopharyngeal nerve transection model, we show that systemic delivery of R-spondin via adenovirus can promote generation of differentiated taste cells despite denervation. Thus, exogenous R-spondin can substitute for neuronal input for taste bud cell replenishment and taste bud maintenance. Using taste organoid cultures, we show that R-spondin is required for generation of differentiated taste cells and that, in the absence of R-spondin in culture medium, taste bud cells are not generated ex vivo. Thus, we propose that R-spondin-2 may be the long-sought neuronal factor that acts on taste stem cells for maintaining taste tissue homeostasis.
Assuntos
Regeneração , Papilas Gustativas/fisiologia , Trombospondinas/metabolismo , Animais , Diferenciação Celular , Camundongos , Organoides , Papilas Gustativas/citologiaRESUMO
"Taste-like" tuft cells in the intestine trigger type 2 immunity in response to worm infection. The secretion of interleukin-13 (IL-13) from type 2 innate lymphoid cells (ILC2) represents a key step in the tuft cell-ILC2 cell-intestinal epithelial cell circuit that drives the clearance of worms from the gut via type 2 immune responses. Hallmark features of type 2 responses include tissue remodeling, such as tuft and goblet cell expansion, and villus atrophy, yet it remains unclear if additional molecular changes in the gut epithelium facilitate the clearance of worms from the gut. Using gut organoids, we demonstrated that IL-4 and IL-13, two type 2 cytokines with similar functions, not only induced the classical type 2 responses (e.g., tuft cell expansion) but also drastically up-regulated the expression of gasdermin C genes (Gsdmcs). Using an in vivo worm-induced type 2 immunity model, we confirmed the up-regulation of Gsdmcs in Nippostrongylus brasiliensis-infected wild-type C57BL/6 mice. Consistent with gasdermin family members being principal effectors of pyroptosis, overexpression of Gsdmc2 in human embryonic kidney 293 (HEK293) cells triggered pyroptosis and lytic cell death. Moreover, in intestinal organoids treated with IL-4 or IL-13, or in wild-type mice infected with N. brasiliensis, lytic cell death increased, which may account for villus atrophy observed in worm-infected mice. Thus, we propose that the up-regulated Gsdmc family may be major effectors for type 2 responses in the gut and that Gsdmc-mediated pyroptosis may provide a conduit for the release of antiparasitic factors from enterocytes to facilitate the clearance of worms.
Assuntos
Morte Celular , Proteínas de Ligação a DNA/metabolismo , Enterócitos/patologia , Imunidade Inata/imunologia , Intestino Delgado/patologia , Infecções por Strongylida/complicações , Células Th2/imunologia , Animais , Proliferação de Células , Proteínas de Ligação a DNA/genética , Enterócitos/imunologia , Enterócitos/metabolismo , Enterócitos/parasitologia , Feminino , Interleucina-13/metabolismo , Interleucina-4/metabolismo , Intestino Delgado/imunologia , Intestino Delgado/metabolismo , Intestino Delgado/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nippostrongylus/fisiologia , Transdução de Sinais , Infecções por Strongylida/imunologia , Infecções por Strongylida/metabolismo , Infecções por Strongylida/parasitologiaRESUMO
Angiogenesis is a critical aspect of wound healing. We investigated the role of keratinocytes in promoting angiogenesis in mice with lineage-specific deletion of the transcription factor FOXO1. The results indicate that keratinocyte-specific deletion of Foxo1 reduces VEGFA expression in mucosal and skin wounds and leads to reduced endothelial cell proliferation, reduced angiogenesis, and impaired re-epithelialization and granulation tissue formation. In vitro FOXO1 was needed for VEGFA transcription and expression. In a porcine dermal wound-healing model that closely resembles healing in humans, local application of a FOXO1 inhibitor reduced angiogenesis. This is the first report that FOXO1 directly regulates VEGFA expression and that FOXO1 is needed for normal angiogenesis during wound healing. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Assuntos
Proteína Forkhead Box O1/metabolismo , Fatores de Transcrição Forkhead/metabolismo , Gengiva/metabolismo , Mucosa Bucal/metabolismo , Neovascularização Fisiológica , Pele/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Cicatrização , Ferimentos e Lesões/metabolismo , Animais , Linhagem Celular , Modelos Animais de Doenças , Feminino , Proteína Forkhead Box O1/deficiência , Proteína Forkhead Box O1/genética , Fatores de Transcrição Forkhead/genética , Gengiva/lesões , Gengiva/patologia , Humanos , Queratinócitos/metabolismo , Queratinócitos/patologia , Masculino , Camundongos Knockout , Mucosa Bucal/lesões , Mucosa Bucal/patologia , Transdução de Sinais , Pele/lesões , Pele/patologia , Suínos , Porco Miniatura , Fator A de Crescimento do Endotélio Vascular/genética , Ferimentos e Lesões/genética , Ferimentos e Lesões/patologiaRESUMO
Grooming, as an evolutionarily conserved repetitive behavior, is common in various animals, including humans, and serves essential functions including, but not limited to, hygiene maintenance, thermoregulation, de-arousal, stress reduction, and social behaviors. In rodents, grooming involves a patterned and sequenced structure, known as the syntactic chain with four phases that comprise repeated stereotyped movements happening in a cephalocaudal progression style, beginning from the nose to the face, to the head, and finally ending with body licking. The context-dependent occurrence of grooming behavior indicates its adaptive significance. This review briefly summarizes the neural substrates responsible for rodent grooming behavior and explores its relevance in rodent models of neuropsychiatric disorders and neurodegenerative diseases with aberrant grooming phenotypes. We further emphasize the utility of rodent grooming as a reliable measure of repetitive behavior in neuropsychiatric models, holding promise for translational psychiatry. Herein, we mainly focus on rodent self-grooming. Allogrooming (grooming being applied on one animal by its conspecifics via licking or carefully nibbling) and heterogrooming (a form of grooming behavior directing towards another animal, which occurs in other contexts, such as maternal, sexual, aggressive, or social behaviors) are not covered due to space constraints.
RESUMO
Taste bud cells are renewed throughout life in a process requiring innervation. Recently, we reported that R-spondin substitutes for neuronal input for taste cell regeneration. R-spondin amplifies WNT signaling by interacting with stem-cell-expressed E3 ubiquitin ligases RNF43/ZNRF3 (negative regulators of WNT signaling) and G-protein-coupled receptors LGR4/5/6 (positive regulators of WNT signaling). Therefore, we hypothesized that RNF43/ZNRF3 may serve as a brake, controlled by gustatory neuron-produced R-spondin, for regulating taste tissue homeostasis. Here, we show that mice deficient for Rnf43/Znrf3 in KRT5-expressing epithelial stem/progenitor cells (RZ dKO) exhibited taste cell hyperplasia; in stark contrast, epithelial tissue on the tongue degenerated. WNT signaling blockade substantially reversed all these effects in RZ dKO mice. Furthermore, innervation becomes dispensable for taste cell renewal in RZ dKO mice. We thus demonstrate important but distinct functions of RNF43/ZNRF3 in regulating taste versus lingual epithelial tissue homeostasis.
Assuntos
Epitélio/metabolismo , Língua/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Animais , Benzenoacetamidas/farmacologia , Nervo Glossofaríngeo/cirurgia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Piridinas/farmacologia , Células-Tronco/citologia , Células-Tronco/metabolismo , Paladar/fisiologia , Papilas Gustativas/metabolismo , Língua/citologia , Ubiquitina-Proteína Ligases/deficiência , Ubiquitina-Proteína Ligases/genética , Via de Sinalização Wnt/efeitos dos fármacosRESUMO
We used multilocus PCR and electrospray ionization mass spectrometry (PCR/ESI-MS) to determine the genotype and drug resistance profiles for 96 Mycobacterium tuberculosis isolates circulating in regions of high and low tuberculosis (TB) endemicity in China. The dominant principal genetic group (PGG) circulating in China was PGG1, and drug-resistant gene mutations were more diversified in the region of low rather than high TB endemicity.
Assuntos
Tipagem Molecular/métodos , Mycobacterium tuberculosis/química , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Tuberculose Resistente a Múltiplos Medicamentos/epidemiologia , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Antituberculosos/farmacologia , Proteínas de Bactérias/genética , China/epidemiologia , DNA Bacteriano/genética , Farmacorresistência Bacteriana , Doenças Endêmicas , Genótipo , Humanos , Mutação de Sentido Incorreto , Mycobacterium tuberculosis/isolamento & purificaçãoRESUMO
Fracture healing is a multistage process characterized by inflammation, cartilage formation, bone deposition, and remodeling. Chondrocytes are important in producing cartilage that forms the initial anlagen for the hard callus needed to stabilize the fracture site. We examined the role of FOXO1 by selective ablation of FOXO1 in chondrocytes mediated by Col2α1 driven Cre recombinase. Experimental mice with lineage-specific FOXO1 deletion (Col2α1Cre+FOXO1L/L) and negative control littermates (Col2α1Cre-FOXO1L/L) were used for in vivo, closed fracture studies. Unexpectedly, we found that in the early phases of fracture healing, FOXO1 deletion significantly increased the amount of cartilage formed, whereas, in later periods, FOXO1 deletion led to a greater loss of cartilage. FOXO1 was functionally important as its deletion in chondrocytes led to diminished bone formation on day 22. Mechanistically, the early effects of FOXO1 deletion were linked to increased proliferation of chondrocytes through enhanced expression of cell cycle genes that promote proliferation and reduced expression of those that inhibit it and increased expression of cartilage matrix genes. At later time points experimental mice with FOXO1 deletion had greater loss of cartilage, enhanced formation of osteoclasts, increased IL-6 and reduced numbers of M2 macrophages. These results identify FOXO1 as a transcription factor that regulates chondrocyte behavior by limiting the early expansion of cartilage and preventing rapid cartilage loss at later phases.
Assuntos
Condrócitos , Consolidação da Fratura , Animais , Calo Ósseo , Cartilagem , Proteína Forkhead Box O1/genética , Camundongos , OsteoclastosRESUMO
Chondrocytes play an essential role in fracture healing by producing cartilage, which forms an anlage for endochondral ossification that stabilizes the healing fracture callus. More recently it has been appreciated that chondrocytes have the capacity to produce factors that may affect the healing process. We examined the role of chondrocytes in angiogenesis during fracture healing and the role of the transcription factor forkhead box-O 1 (FOXO1), which upregulates wound healing in soft tissue. Closed fractures were induced in experimental mice with lineage-specific FOXO1 deletion by Cre recombinase under the control of a collagen-2α1 promoter element (Col2α1Cre+ FOXO1L/L ) and Cre recombinase negative control littermates containing flanking loxP sites (Col2α1Cre- FOXO1L/L ). Experimental mice had significantly reduced CD31+ new vessel formation. Deletion of FOXO1 in chondrocytes in vivo suppressed the expression of vascular endothelial growth factor-A (VEGFA) at both the protein and mRNA levels. Overexpression of FOXO1 in chondrocytes in vitro increased VEGFA mRNA levels and VEGFA transcriptional activity whereas silencing FOXO1 reduced it. Moreover, FOXO1 interacted directly with the VEGFA promoter and a deacetylated FOXO1 mutant enhanced VEGFA expression whereas an acetylated FOXO1 mutant did not. Lastly, FOXO1 knockdown by siRNA significantly reduced the capacity of chondrocytes to stimulate microvascular endothelial cell tube formation in vitro. The results indicate that chondrocytes play a key role in angiogenesis which is FOXO1 dependent and that FOXO1 in chondrocytes regulates a potent angiogenic factor, VEGFA. These studies provide new insight into fracture healing given the important role of vessel formation in the fracture repair process. © 2018 American Society for Bone and Mineral Research.
Assuntos
Condrócitos/metabolismo , Proteína Forkhead Box O1/metabolismo , Consolidação da Fratura , Neovascularização Fisiológica , Fator A de Crescimento do Endotélio Vascular/biossíntese , Animais , Linhagem Celular , Colágeno Tipo II/biossíntese , Colágeno Tipo II/genética , Regulação para Baixo , Células Endoteliais/patologia , Proteína Forkhead Box O1/genética , Deleção de Genes , Camundongos , Camundongos Transgênicos , Molécula-1 de Adesão Celular Endotelial a Plaquetas/biossíntese , Transcrição Gênica , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
The importance of some putative efflux pumps in conferring drug resistance on certain strain of Mycobacterium tuberculosis was determined by using multidrug-resistant (MDR) strain 1499 bearing defined mutations in rpoB and katG. The clinical isolate 1499 was tested for drug sensitivity and expression level of five putative efflux pump genes by real-time RT-PCR. In the presence of rifampicin or isoniazid, this isolate showed increased transcription of putative efflux pump genes Rv1258c and Rv1410c. Rv1819c was overexpressed only upon isoniazid exposure (p<0.05). Overexpression of some efflux genes is associated with multidrug resistance in certain clinical isolate of M. tuberculosis.
Assuntos
Transportadores de Cassetes de Ligação de ATP/biossíntese , Antituberculosos/farmacologia , Proteínas de Bactérias/biossíntese , Expressão Gênica , Proteínas de Membrana Transportadoras/biossíntese , Mycobacterium tuberculosis/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Alquil e Aril Transferases/biossíntese , Alquil e Aril Transferases/genética , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Farmacorresistência Bacteriana Múltipla/genética , Humanos , Isoniazida/farmacologia , Proteínas de Membrana Transportadoras/genética , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rifampina/farmacologiaRESUMO
Purpose: Myopia induction accompanies increased scleral cyclic adenosine phosphate (cAMP) levels and collagen degradation in mammalian models. We compared the scleral gene expression changes following monocular form deprivation (FD) with those induced by adenylate cyclase activation with forskolin (FSK) in guinea pigs. Methods: Guinea pigs were assigned to FD, FSK-treated, and age-matched (AM) control groups. FSK was injected monocularly into the inferior palpebral subconjunctiva daily for 4 days. After scleral RNA extraction, a gene microarray scanner and software were used to evaluate the gene expression patterns, followed by pathway analysis using Gene Ontology tools. Quantitative PCR (qPCR) was used to analyze the expression of 10 candidate genes in separate sets of form-deprived, vehicle-injected, and AM animals. Results: FSK injections differentially regulated 13 collagen subtypes compared to AM and FD groups. FSK also downregulated Acta2 and Tgf-ß2 compared to the AM eyes. Collagen subtypes and Acta2 underwent larger downregulation in the FSK group than during FD. FSK differentially regulated Rarb, Rxrg, Fzd5, Ctnnd2, Dkk2, and Dkk3, which have been linked to ocular growth. Only a few genes were differentially expressed between the FD and AM groups. There was 80% agreement in the direction of gene regulation between microarray and qPCR results. No significant differences were identified between vehicle-injected and AM eyes. Conclusions: Collagen, a major scleral extracellular matrix component, is degraded during myopia. Given that FSK and FD both promote myopia through increased collagen degradation, targeting cAMP signaling pathway genes could suppress myopia development.
Assuntos
AMP Cíclico/fisiologia , Miopia/metabolismo , Esclera/metabolismo , Privação Sensorial/fisiologia , Animais , Colforsina/farmacologia , Modelos Animais de Doenças , Regulação para Baixo , Colágenos Associados a Fibrilas/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Cobaias , Análise em Microsséries , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Type 1 diabetes impairs fracture healing. We tested the hypothesis that diabetes affects chondrocytes to impair fracture healing through a mechanism that involves the transcription factor FOXO1. Type 1 diabetes was induced by streptozotocin in mice with FOXO1 deletion in chondrocytes (Col2α1Cre+FOXO1L/L) or littermate controls (Col2α1Cre-FOXO1L/L) and closed femoral fractures induced. Diabetic mice had 77% less cartilage and 30% less bone than normoglycemics evaluated histologically and by micro-computed tomography. Both were reversed with lineage-specific FOXO1 ablation. Diabetic mice had a threefold increase in osteoclasts and a two- to threefold increase in RANKL mRNA or RANKL-expressing chondrocytes compared with normoglycemics. Both parameters were rescued by FOXO1 ablation in chondrocytes. Conditions present in diabetes, high glucose (HG), and increased advanced glycation end products (AGEs) stimulated FOXO1 association with the RANKL promoter in vitro, and overexpression of FOXO1 increased RANKL promoter activity in luciferase reporter assays. HG and AGE stimulated FOXO1 nuclear localization, which was reversed by insulin and inhibitors of TLR4, histone deacetylase, nitric oxide, and reactive oxygen species. The results indicate that chondrocytes play a prominent role in diabetes-impaired fracture healing and that high levels of glucose, AGEs, and tumor necrosis factor-α, which are elevated by diabetes, alter RANKL expression in chondrocytes via FOXO1.
Assuntos
Diabetes Mellitus Experimental/metabolismo , Fraturas do Fêmur/metabolismo , Proteína Forkhead Box O1/metabolismo , Consolidação da Fratura/genética , Animais , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Diabetes Mellitus Experimental/genética , Fraturas do Fêmur/genética , Proteína Forkhead Box O1/genética , Consolidação da Fratura/efeitos dos fármacos , Regulação da Expressão Gênica , Glucose/farmacologia , Produtos Finais de Glicação Avançada/farmacologia , Camundongos , Camundongos Knockout , Ligante RANK/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Microtomografia por Raio-XRESUMO
Numerous physiological processes of mammals, including bone metabolism, are regulated by the circadian clock system, which consists of a central regulator, the suprachiasmatic nucleus (SCN), and the peripheral oscillators of the BMAL1/CLOCK-PERs/CRYs system. Various bone turnover markers and bone metabolism-regulating hormones such as melatonin and parathyroid hormone (PTH) display diurnal rhythmicity. According to previous research, disruption of the circadian clock due to shift work, sleep restriction, or clock gene knockout is associated with osteoporosis or other abnormal bone metabolism, showing the importance of the circadian clock system for maintaining homeostasis of bone metabolism. Moreover, common causes of osteoporosis, including postmenopausal status and aging, are associated with changes in the circadian clock. In our previous research, we found that agonism of the circadian regulators REV-ERBs inhibits osteoclast differentiation and ameliorates ovariectomy-induced bone loss in mice, suggesting that clock genes may be promising intervention targets for abnormal bone metabolism. Moreover, osteoporosis interventions at different time points can provide varying degrees of bone protection, showing the importance of accounting for circadian rhythms for optimal curative effects in clinical treatment of osteoporosis. In this review, we summarize current knowledge about circadian rhythms and bone metabolism.
Assuntos
Osso e Ossos/metabolismo , Osso e Ossos/fisiologia , Ritmo Circadiano/fisiologia , Animais , Relógios Circadianos/fisiologia , Homeostase/fisiologia , Humanos , Melatonina/metabolismo , Osteoporose/metabolismo , Osteoporose/fisiopatologia , Pós-Menopausa/metabolismo , Pós-Menopausa/fisiologiaRESUMO
Neural physiological functions and synaptic changes underlying the pathogenesis of depression have obtained great achievements. However, neuronal morphological changes under a depressive state have not been well understood yet. Here a depressive-like YFP-H transgenic mouse model was produced by light deprivation (LD), and morphological changes of retinal ganglion cells (RGCs) and primary visual and auditory cortical layer 5 pyramidal cells (L5PCs) were investigated. Three distinct RGC subtypes were identified based on soma- and dendritic field (DF) size. RGA cells were highlighted by large soma and medium-sized to large DF. RGB cells were characterized by small- to medium-sized soma and small- to medium-sized DF. RGC cells were typical of small- to medium-sized soma and large DF. LD showed cell-type-specific morphological orchestrations on RGCs and predominantly promoted the dendritic growth of RGA cells, leaving no significant effect on RGB and RGC cells. LD produced a consistently suppressed effect on the morphology of primary visual and auditory cortical L5PCs. LD enhanced the dendritic spine density of primary visual cortical L5PCs, implying a compensation mechanism underlying morphological changes in individual cortical L5PCs. The increased morphological complexity of RGA cells and the simplified morphology of cortical L5PCs suggest a broad range of neuronal morphological "cross-modal plasticity" among different brain areas. Our observations in morphological changes of RGCs and cortical L5PCs under a depressive-like state will provide some insights into the pathogenesis of depression at a single neuronal morphological level.
Assuntos
Luz , Plasticidade Neuronal/fisiologia , Células Piramidais/citologia , Células Ganglionares da Retina/citologia , Privação Sensorial/fisiologia , Animais , Proteínas de Bactérias/genética , Dendritos/fisiologia , Espinhas Dendríticas/fisiologia , Proteínas Luminescentes/genética , Camundongos , Camundongos TransgênicosRESUMO
BACKGROUND: Cytokines play an important role in cell-mediated immune responses against Mycobacterium tuberculosis (Mtb) infection. Cytokine profile specifically associated with active tuberculosis (ATB) patients, subjects with latent tuberculosis infection (LTBI) and non-infected individuals remains to be determined. METHODS: We enrolled a total of 92 subjects including patients with ATB (n = 25), LTBI (n = 36) and healthy controls (HC, n = 31) to investigate the cytokine production by peripheral blood mononuclear cells after Mtb purified protein derivative (PPD) stimulation which was evaluated by a beads-based multiplex assay system. RESULTS: The production of IL-1ß, IL-2, IL-6, IL-10, IL-17, G-CSF, IFN-γ, IP-10, MIP-1α and TNF-α was abundantly induced by PPD in all three groups. The levels of IL-2, IL-10, IFN-γ, IP-10 and TNF-α were significantly higher in LTBI group than in ATB group. The combination of PPD-stimulated IL-2 and IL-10 accurately identified 84.0% of ATB and 88.9% of LTBI. We validated the use of PPD-stimulated IL-2 and IL-10 test combined with T-SPOT.TB test in a cohort of 44 subjects with TB suspicion. The sensitivity and specificity of the combined test were 83.3% and 92.3%, respectively. The PPD-stimulated IL-2/IFN-γ ratio (p < 0.001) in LTBI subjects was significantly higher than in active TB patients. CONCLUSION: Our study identified cytokine patterns characteristic of ATB and LTBI. Cytokines such as IL-2 and IL-10 may serve as biomarkers for distinguishing ATB from LTBI and healthy control and may contribute to intervention and improvement in TB diagnosis.
Assuntos
Citocinas/biossíntese , Tuberculose/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/metabolismo , Estudos de Casos e Controles , Células Cultivadas , Criança , Pré-Escolar , Diagnóstico Diferencial , Feminino , Humanos , Testes de Liberação de Interferon-gama , Tuberculose Latente/diagnóstico , Tuberculose Latente/imunologia , Leucócitos Mononucleares/imunologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Tuberculina/imunologia , Teste Tuberculínico , Tuberculose/imunologiaRESUMO
Neutrophils play an essential role in the innate immune response to microbial infection and are particularly important in clearing bacterial infection. We investigated the role of the transcription factor FOXO1 in the response of neutrophils to bacterial challenge with Porphyromonas gingivalis in vivo and in vitro. In these experiments, the effect of lineage-specific FOXO1 deletion in LyzM.Cre+FOXO1L/L mice was compared with matched littermate controls. FOXO1 deletion negatively affected several critical aspects of neutrophil function in vivo including mobilization of neutrophils from the bone marrow (BM) to the vasculature, recruitment of neutrophils to sites of bacterial inoculation, and clearance of bacteria. In vitro FOXO1 regulated neutrophil chemotaxis and bacterial killing. Moreover, bacteria-induced expression of CXCR2 and CD11b, which are essential for several aspects of neutrophil function, was dependent on FOXO1 in vivo and in vitro. Furthermore, FOXO1 directly interacted with the promoter regions of CXCR2 and CD11b. Bacteria-induced nuclear localization of FOXO1 was dependent upon toll-like receptor (TLR) 2 and/or TLR4 and was significantly reduced by inhibitors of reactive oxygen species (ROS and nitric oxide synthase) and deacetylases (Sirt1 and histone deacetylases). These studies show for the first time that FOXO1 activation by bacterial challenge is needed to mobilize neutrophils to transit from the BM to peripheral tissues in response to infection as well as for bacterial clearance in vivo. Moreover, FOXO1 regulates neutrophil function that facilitates chemotaxis, phagocytosis, and bacterial killing.
RESUMO
Light sensory experience plays a crucial role in the regulation of mood, and light deficiency is considered as one important factor potentially leading to depression. Women are twice as likely as men to suffer from depression. However, the physiological mechanism underlying sex differences in the prevalence, incidence and morbidity risk of depression is still poorly understood. The potential causal relationship between sex dimorphic behavioral deficits and altered intrinsic electrophysiological properties of Layer V pyramidal cells (L5PCs) in the motor cortex was investigated using a mouse model with depression-like behavior that was induced by light deprivation. The depression-like behavior was characterized by increased immobility and decreased activity in the forced swimming test and tail suspension test. Compared with male depressive-like mice, light deprivation (LD) induced longer immobile behavior while shorter active behavior in female depressive-like mice, indicating that LD produces a sexual dimorphic effect on depression-like behavior with more severe depressive-like symptoms in females. LD induced lower locomotor activity in female depressive-like mice as evidenced by the significant decrease in pole-climbing and swimming during the anti-static fatigue test and exhaustive swimming test correspondingly. LD also significantly decreased the intrinsic excitability of L5PCs in female depressive-like mice, which may explain the reduced active behavior and locomotor activity of female mice. Collectively, it indicates that LD produces a sexual dimorphic effect on the depression-like behavior, locomotor activity and neural excitability in mice, and may suggest a causal relationship between the more severe depressive conditions and decreased neural excitability of L5PCs in female mice. These divergent findings from male and female depressive-like mice may provide one potential route to the physiological mechanism underlying sex differences in the prevalence of depression at a level of single neurons.
Assuntos
Depressão/psicologia , Luz , Atividade Motora , Córtex Motor/fisiologia , Animais , Escuridão/efeitos adversos , Depressão/etiologia , Depressão/fisiopatologia , Feminino , Masculino , Camundongos Endogâmicos ICR , Células Piramidais/fisiologia , Caracteres SexuaisRESUMO
The development of myopia is associated with decreased ocular scleral collagen synthesis in humans and animal models. Collagen synthesis is, in part, under the influence of cyclic adenosine monophosphate (cAMP). We investigated the associations between cAMP, myopia development in guinea pigs, and collagen synthesis by human scleral fibroblasts (HSFs). Form-deprived myopia (FDM) was induced by unilateral masking of guinea pig eyes. Scleral cAMP levels increased selectively in the FDM eyes and returned to normal levels after unmasking and recovery. Unilateral subconjunctival treatment with the adenylyl cyclase (AC) activator forskolin resulted in a myopic shift accompanied by reduced collagen mRNA levels, but it did not affect retinal electroretinograms. The AC inhibitor SQ22536 attenuated the progression of FDM. Moreover, forskolin inhibited collagen mRNA levels and collagen secretion by HSFs. The inhibition was reversed by SQ22536. These results demonstrate a critical role of cAMP in control of myopia development. Selective regulation of cAMP to control scleral collagen synthesis may be a novel therapeutic strategy for preventing and treating myopia.
Assuntos
Colágeno/metabolismo , AMP Cíclico/metabolismo , Miopia/metabolismo , Esclera/metabolismo , Animais , Colforsina/efeitos adversos , Colforsina/farmacologia , Colágeno/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Cobaias , Pressão Intraocular/efeitos dos fármacos , Miopia/induzido quimicamente , Miopia/genética , Retina/efeitos dos fármacos , Retina/metabolismo , Privação SensorialRESUMO
Antigens encoded in the region of difference (RD) of Mycobacterium tuberculosis constitute a potential source of specific immunodiagnostic antigens for distinguishing tuberculosis (TB) infection from BCG vaccination. We evaluated the diagnostic potential of specific T-cell epitopes selected from two immunodominant antigens, Rv1985c and Rv3425, from RD2 and RD11, respectively, on the basis of epitope mapping, in TB patients and BCG-vaccinated healthy individuals. Using a whole-blood gamma interferon release assay, a wide array of epitopes was recognized on both Rv1985c and Rv3425 in TB patients. Those epitopes that could specifically discriminate TB infection from BCG vaccination were carefully selected, and the most promising peptide pools from Rv1985c showed a sensitivity of 53.9% and a specificity of 95.5%. When the novel specific peptides from Rv1985c joined the diagnostic antigens in the QuantiFERON-TB Gold In-Tube (QFT-IT) assay, the sensitivity was increased from 86.4% to 96.2%, with no drop in specificity. These results indicate that the peptide pools selected from Rv1985c and Rv3425 have the potential to diagnose TB infection by a method that may be routinely used in clinical laboratories.
Assuntos
Antígenos de Bactérias , Vacina BCG/imunologia , Testes de Liberação de Interferon-gama/métodos , Mycobacterium tuberculosis/imunologia , Tuberculose/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos de Bactérias/imunologia , Diagnóstico Diferencial , Epitopos de Linfócito T/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Tuberculose/imunologia , Adulto JovemRESUMO
BACKGROUND: The Mycobacterium tuberculosis (Mtb)-specific T-cell interferon gamma release assays (IGRAs) are useful in detecting Mtb infection but perform poorly at distinguishing active tuberculosis disease (ATB) and latent tuberculosis infection (LTBI). This study is aimed at evaluating additional cytokines as biomarkers besides interferon-gamma (IFN-γ) to improve the identification of ATB and LTBI. METHODOLOGY/PRINCIPAL FINDINGS: Sixty-six patients with ATB, 73 household contacts (HHC) of ATB patients and 76 healthy controls (HC) were recruited to undergo QuantiFERON TB GOLD in-tube assay (QFT) and the enzyme-linked immunosorbent assay (ELISA) where the release of IFN-γ, IFN-γ inducible protein 10 (IP-10), Interleukin 2 (IL-2) and Tumor Necrosis Factor-α (TNF-α) was determined in the whole blood with or without antigen-stimulation. The positive rates of the QFT, IP-10 and IL-2 tests were 86.4%, 89.4% and 86.4% for the ATB group with no difference between them (p>0.05). However, QFT in combination with IP-10 and IL-2 significantly increased the detection rate to 95.5% in the ATB group (pâ=â0.03) and the indeterminate rate of all samples decreased from 2.3% (5/215) to 0.4% (1/215). The un-stimulated level of IP-10 was significantly higher in the HHC than the ATB and HC groups. The IP-10 responses were strongly associated with extended Mtb exposure time and the degree of smear-positivity of the index cases. The IL-2/IFN-γ ratio in the antigen-stimulated plasma could discriminate LTBI from ATB with a sensitivity of 77.2% and a specificity of 87.2%. CONCLUSION: The increased Mtb-specific antigen-stimulated expression of IP-10 and IL-2 may be useful for detecting both ATB and LTBI. Combining the QFT with IP-10 and IL-2 could increase the detection accuracy of active TB over the QFT alone.
Assuntos
Vacina BCG/metabolismo , Quimiocina CXCL10/sangue , Interleucina-2/sangue , Tuberculose Latente/sangue , Tuberculose/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos/metabolismo , Biomarcadores/metabolismo , Estudos Transversais , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Interferon gama/metabolismo , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Linfócitos T/metabolismo , Fatores de Tempo , Teste Tuberculínico , Fator de Necrose Tumoral alfa/metabolismoRESUMO
To explore biologic behaviors and disease relevance of microRNAs (miRNAs) in the development of active tuberculosis (ATB), we investigated the expression profile of Mycobacterium tuberculosis (MTB) purified protein derivative (PPD)-induced miRNAs to determine the specific miRNAs involved in the pathogenesis of ATB. The expression profile of miRNA under PPD challenge was first measured using microarray analysis in peripheral blood mononuclear cells isolated from ATB patients and healthy controls (HC). The remarkably reactive miRNAs were then validated in a larger cohort by quantitative real-time polymerase chain reaction (qRT-PCR). The receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic value of the determined PPD-responsive miRNAs. The potential targets for those miRNAs were also predicted by computational programs. Fourteen of 866 human miRNAs exhibited at least 1.8-fold difference in the ratio of expression level before and after stimulation with PPD between the ATB and HC groups. The qRT-PCR study validated the findings from microarray-based screening, in which miR-155 exhibited a fold change of 1.4 in the HC group and 3.7 in the ATB group upon PPD stimulation (p < 0.0001); miR-155* exhibited a fold change of 1.9 in the HC and 4.6 in the ATB group (p < 0.005). In ROC plots, the area under the curve was 0.8972 for miR-155 and 0.7945 for miR-155*. The background expression of these 2 microRNAs exhibited no differences between the ATB and HC groups. miR-155 and miR-155* exhibited characteristic expression by TB-specific antigen, suggesting that they can be potential diagnostic markers under the challenge of specific MTB antigens.