RESUMEN
T cell polyfunctionality is a hallmark of protective immunity against pathogens and cancer, yet the molecular mechanism governing it remains mostly elusive. We found that canonical Wnt agonists inhibited human memory CD8+ T cell differentiation while simultaneously promoting the generation of highly polyfunctional cells. Downstream effects of Wnt activation persisted after removal of the drug, and T cells remained polyfunctional following subsequent cell division, indicating the effect is epigenetically regulated. Wnt activation induced a gene expression pattern that is enriched with stem cell-specific gene signatures and upregulation of protein arginine methyltransferase 1 (PRMT1), a known epigenetic regulator. PRMT1+CD8+ T cells are associated with enhanced polyfunctionality, especially the ability to produce IL-2. In contrast, inhibition of PRMT1 ameliorated the effects of Wnt on polyfunctionality. Chromatin immunoprecipitation revealed that H4R3me2a, a permissive transcription marker mediated by PRMT1, increased at the IL-2 promoter loci following Wnt activation. In vivo, Wnt-treated T cells exhibited superior polyfunctionality and persistence. When applied to cytomegalovirus (CMV) donor-seropositive, recipient-seronegative patients (D+/R-) lung transplant patient samples, Wnt activation enhanced CMV-specific T cell polyfunctionality, which is important in controlling CMV diseases. These findings reveal a molecular mechanism governing T cell polyfunctionality and identify PRMT1 as a potential target for T cell immunotherapy.
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Linfocitos T CD8-positivos/inmunología , Epigénesis Genética/inmunología , Células T de Memoria/inmunología , Proteína-Arginina N-Metiltransferasas/inmunología , Proteínas Represoras/inmunología , Vía de Señalización Wnt/inmunología , Humanos , Interleucina-2/inmunología , Trasplante de Pulmón , Proteínas Wnt/inmunologíaRESUMEN
Overexpression of WLS, an upstream protein in the Wnt pathway, has been implicated in several non-osteogenic tumours. This study represents the first attempt at evaluating WLS expression in various bone and soft tissue tumours using YJ5, a monoclonal antibody specific to WLS, with the aim of elucidating its utility in discerning tumours with aberrant Wnt signalling and as a marker of osteogenic lineage in challenging cases. Tumour tissue sections of 144 bone mass lesions and 63 soft tissue mass lesions were immunostained with the YJ5 antibody following standardised protocols. Subsequent assessment of immunoreactivity segregated cases into one of three groups: absent/weak, moderate, or strong YJ5 immunoreactivity. For the bone tumours, strong YJ5 immunoreactivity was seen in almost all osteosarcomas and chondroblastomas, all osteoblastomas and osteoid osteomas. In contrast, all other cartilaginous tumours, chordomas, aneurysmal bone cysts, chondromyxoid fibromas, most fibrous dysplasias and most giant cell tumours exhibited absent/weak YJ5 immunostaining. For the soft tissue tumours, a more heterogeneous pattern of YJ5 immunoreactivity was observed. Because diffuse and strong YJ5 expression is identified in almost all benign and malignant bone tumours with osteoblastic activity, it can be potentially utilised as an immunohistochemical marker to support osteogenic lineage. If interpreted in the appropriate context, this marker is useful in determining whether a malignant bone tumour is an osteosarcoma, particularly in those subtypes with no or minimal osteoid or unusual morphological features. This marker can also complement SATB2 to denote osteogenic lineage.
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Anticuerpos Monoclonales , Péptidos y Proteínas de Señalización Intracelular , Osteosarcoma , Receptores Acoplados a Proteínas G , Biomarcadores de Tumor/metabolismo , Neoplasias Óseas/patología , Huesos/patología , Humanos , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intracelular/inmunología , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de Unión a la Región de Fijación a la Matriz/análisis , Proteínas de Unión a la Región de Fijación a la Matriz/metabolismo , Osteosarcoma/diagnóstico , Osteosarcoma/patología , Receptores Acoplados a Proteínas G/inmunología , Receptores Acoplados a Proteínas G/metabolismo , Estudios Retrospectivos , Neoplasias de los Tejidos Blandos/patología , Factores de Transcripción/análisis , Factores de Transcripción/metabolismo , Proteínas Wnt/inmunología , Proteínas Wnt/metabolismo , Vía de Señalización WntRESUMEN
CD8+ T cells do not rely solely on cytotoxic functions for significant HIV control. Moreover, the noncytotoxic CD8+ T cell antiviral response is a primary mediator of natural HIV control such as that seen in HIV elite controllers and long-term nonprogressors that does not require combined antiretroviral therapy. In this study, we investigated the biological factors contributing to the noncytotoxic control of HIV replication mediated by primary human CD8+ T cells. We report that canonical Wnt signaling inhibits HIV transcription in an MHC-independent, noncytotoxic manner and that mediators of this pathway correlate with HIV controller clinical status. We show that CD8+ T cells express all 19 Wnts and CD8+ T cell-conditioned medium (CM) induced canonical Wnt signaling in infected recipient cells while simultaneously inhibiting HIV transcription. Antagonizing canonical Wnt activity in CD8+ T cell CM resulted in increased HIV transcription in infected cells. Further, Wnt2b expression was upregulated in HIV controllers versus viremic patients, and in vitro depletion of Wnt2b and/or Wnt9b from CD8+ CM reversed HIV inhibitory activity. Finally, plasma concentration of Dkk-1, an antagonist of canonical Wnt signaling, was higher in viremic patients with lower CD4 counts. This study demonstrates that canonical Wnt signaling inhibits HIV and significantly correlates with HIV controller status.
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Linfocitos T CD8-positivos , Regulación de la Expresión Génica/inmunología , Glicoproteínas , Infecciones por VIH , VIH-1 , Inmunidad Celular , Proteínas Wnt , Vía de Señalización Wnt/inmunología , Adulto , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/patología , Femenino , Glicoproteínas/sangre , Glicoproteínas/inmunología , Infecciones por VIH/sangre , Infecciones por VIH/inmunología , Infecciones por VIH/patología , VIH-1/inmunología , VIH-1/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intercelular/sangre , Péptidos y Proteínas de Señalización Intercelular/inmunología , Masculino , Proteínas Wnt/sangre , Proteínas Wnt/inmunologíaRESUMEN
Root resorption is usually inflammatory in nature and has a tight link with immune system. Intermittent parathyroid hormone (iPTH) could promote cementum regeneration. The cross-talk of immune cells and cementoblasts may play an important role in the regeneration which stayed to be elucidated. In this study, a CD8+ T cells-OCCM-30 cells coculture system was established in vitro to investigate whether CD8+ T cells could enhance the anabolic effect of iPTH on cementoblasts and to find out the potential link of the effect with Wnt signal pathway. Determined by real-time PCR and Western Blot, we found an amplified cementogenesis in the OCCM-30 cells from coculture system, including increased mRNA and protein expression of Alp, Opn and Runx2, ALP activity and mineralization. We also found iPTH could increase the expression of Wnt10b in CD8+ T cells by ELISA. In addition, Wnt10b would promote the proliferation of OCCM-30 cells, while the effect on differentiation was various in different culture medium. These results demonstrated that the stimulating effect of iPTH on cementoblasts could be mediated through an interaction with CD8+ T cells, and T-cell-induced Wnt10b might be a key mechanism in the mediation.
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Anabolizantes/inmunología , Linfocitos T CD8-positivos/inmunología , Cemento Dental/inmunología , Hormona Paratiroidea/inmunología , Proteínas Wnt/inmunología , Animales , Línea Celular , Técnicas de Cocultivo , Ratones Endogámicos BALB C , Proteínas Wnt/genéticaRESUMEN
The Wnt signaling pathway is a conserved pathway involved in important cellular processes such as the control of embryonic development, cellular polarity, cellular migration, and cell proliferation. In addition to playing a central role during embryogenesis, this pathway is also an essential part of adult homeostasis. Indeed, it controls the proliferation of epithelial cells in different organs such as intestine, lung, and kidney, and guarantees the maintenance of the mucosa in physiological conditions. The origin of this molecular pathway is the binding between Wnt ligands (belonging to a family of 19 different homologous secreted glycoproteins) and their specific membrane receptors, from the Frizzled receptor family. This specific interaction triggers the activation of the signaling cascade, which in turn activates or suppresses the expression of different genes in order to change the behavior of the cell. On the other hand, alterations of this pathway have been described in pathological conditions such as inflammation, fibrosis, and cancer. In recent years, macrophages-among other cell types-have emerged as a potential source of Wnt ligands. Due to their high plasticity, macrophages, which are central to the innate immune response, are capable of adopting different phenotypes depending on their microenvironment. In the past, two different phenotypes were described: a proinflammatory phenotype-M1 macrophages-and an anti-inflammatory phenotype-M2 macrophages-and a selective expression of Wnt ligands has been associated with said phenotypes. However, nowadays it is assumed that macrophages in vivo move through a continual spectrum of functional phenotypes. In both physiological and pathological (inflammation, fibrosis and cancer) conditions, the accumulation and polarization of macrophages conditions the future of the tissue, facilitating various scenarios, such as resolution of inflammation, activation of fibrosis, and cancer development due to the modulation of the Wnt signaling pathway, in autocrine and paracrine manner. In this work, we provide an overview of studies that have explored the role of macrophages and how they act as a source of Wnt ligands and as mediators of mucosal integrity.
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Macrófagos/inmunología , Membrana Mucosa/inmunología , Proteínas Wnt/inmunología , Vía de Señalización Wnt/inmunología , Humanos , Inmunidad Innata/inmunología , Inflamación/inmunología , Inflamación/metabolismo , Ligandos , Macrófagos/clasificación , Macrófagos/metabolismo , Membrana Mucosa/metabolismo , Neoplasias/inmunología , Neoplasias/metabolismo , Unión Proteica , Proteínas Wnt/metabolismoRESUMEN
OBJECTIVE: To investigate the participation of canonical Wnt and NF-κB signaling pathways in an experimental model of chronic arthritis induced by methylated bovine serum albumin (mBSA) in rat temporomandibular joint (TMJ). MATERIALS AND METHODS: Wistar rats were sensitized by mBSA+Complete Freund Adjuvant (CFA)/Incomplete Freund Adjuvant (IFA) on the first 14 days (1 ×/week). Subsequently, they received 1, 2 or 3 mBSA or saline solution injections into the TMJ (1 ×/week). Hypernociceptive threshold was assessed during the whole experimental period. 24 h after the mBSA injections, the TMJs were removed for histopathological and immunohistochemical analyses for TNF-α, IL-1ß, NF-κB, RANKL, Wnt-10b, ß-catenin and DKK1. RESULTS: The nociceptive threshold was significantly reduced after mBSA injections. An inflammatory infiltrate and thickening of the synovial membrane were observed only after mBSA booster injections. Immunolabeling of TNF-α, IL-1ß and Wnt-10b was increased in the synovial membrane in arthritic groups. The immunoexpression of nuclear ß-catenin was significantly higher only in the group that received 2 booster TMJ injections. However, NF-κB, RANKL and DKK1 immunoexpression were increased only in animals with 3 mBSA intra-articular injections. CONCLUSION: Our results suggest that canonical Wnt and NF-κB signaling pathways participate in the hypernociception and inflammatory response in TMJ synovial membrane during the development of rheumatoid arthritis in rats.
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Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Hiperalgesia/inmunología , FN-kappa B/inmunología , Articulación Temporomandibular/inmunología , Vía de Señalización Wnt , Animales , Artritis Experimental/patología , Artritis Reumatoide/patología , Adyuvante de Freund , Hiperalgesia/patología , Péptidos y Proteínas de Señalización Intercelular/inmunología , Interleucina-1beta/inmunología , Lípidos , Masculino , Ligando RANK/inmunología , Ratas Wistar , Albúmina Sérica Bovina , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Articulación Temporomandibular/patología , Factor de Necrosis Tumoral alfa/inmunología , Proteínas Wnt/inmunologíaRESUMEN
BACKGROUND/AIMS: Current drug therapies for osteoarthritis (OA) are not practical because of the cytotoxicity and severe side-effects associated with most of them. Artemisinin (ART), an antimalarial agent, is well known for its safety and selectivity to kill injured cells. Based on its anti-inflammatory activity and role in the inhibition of OA-associated Wnt/ß-catenin signaling pathway, which is crucial in the pathogenesis of OA, we hypothesized that ART might have an effect on OA. METHODS: The chondro-protective and antiarthritic effects of ART on interleukin-1-beta (IL-1ß)-induced and OA patient-derived chondrocytes were investigated in vitro using cell viability assay, glycosaminoglycan secretion, immunofluorescence, quantitative reverse transcription-polymerase chain reaction, and western blotting. We also used OA model rats constructed by anterior cruciate ligament transection and medial meniscus resection (ACLT+MMx) in the joints to investigate the effects of ART on OA by gross observation, morphological staining, immunohistochemistry, and enzyme-linked immunosorbent assay. RESULTS: ART exhibited potent anti-inflammatory effects by inhibiting the expression of proinflammatory chemokines and cytokines, including interleukin (IL)-1ß, IL-6, tumor necrosis factor alpha, and matrix metallopeptidase-13. It also showed favorable chondro-protective effect as evidenced by enhanced cell proliferation and viability, increased glycosaminoglycan deposition, prevention of chondrocyte apoptosis, and degeneration of cartilage. Further, ART inhibited OA progression and cartilage degradation via the Wnt/ß-catenin signaling pathway, suggesting that it might serve as a Wnt/ß-catenin antagonist to reduce inflammation and prevent cartilage degradation. CONCLUSION: In conclusion, ART alleviates IL-1ß-mediated inflammatory response and OA progression by regulating the Wnt/ß-catenin signaling pathway. Thereby, it might be developed as a potential therapeutic agent for OA.
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Antiinflamatorios/uso terapéutico , Artemisininas/uso terapéutico , Condrocitos/efectos de los fármacos , Osteoartritis/tratamiento farmacológico , Vía de Señalización Wnt/efectos de los fármacos , Adulto , Anciano , Animales , Antiinflamatorios/farmacología , Antimaláricos/farmacología , Antimaláricos/uso terapéutico , Artemisininas/farmacología , Células Cultivadas , Condrocitos/inmunología , Condrocitos/patología , Femenino , Humanos , Interleucina-1beta/inmunología , Masculino , Persona de Mediana Edad , Osteoartritis/inmunología , Osteoartritis/patología , Ratas Sprague-Dawley , Proteínas Wnt/inmunología , Adulto JovenRESUMEN
Despite the dramatic antitumor efficiency of certain immune checkpoint inhibitors, immunotherapy has met a bottleneck regarding the response rate and resistance in cancer patients. Increasing evidence indicates that Wnt/ß-catenin signaling, one of the best-characterized cancer drivers, promotes cancer progression by regulating the tumor-immune cycle in most of the nodes, including dendritic cells, T cells, and tumor cells. Specifically, abnormal Wnt/ß-catenin signaling directly alters a number of regulators critical for the antitumor activities of T cells, especially effector T cells, T helper cells, and regulatory T cells. We propose that targeting Wnt/ß-catenin signaling would potentially improve clinical outcomes of cancer patients by overcoming the primary, adaptive, and acquired resistance to immunotherapy.
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Inmunoterapia/métodos , Neoplasias/inmunología , Neoplasias/terapia , Vía de Señalización Wnt/inmunología , Animales , Linfocitos T CD8-positivos/inmunología , Células Dendríticas/inmunología , Humanos , Interleucina-1/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Reguladores/inmunología , Proteínas Wnt/inmunología , beta Catenina/inmunologíaRESUMEN
The Wnt gene family encodes secreted glycoproteins involved in a wide variety of biological processes, including embryo development, cell proliferation and differentiation, and tissue regeneration. The Wnt pathway exists in all metazoan animals, however, the relevant research is rare in crustaceans. Here we described 12 Wnt genes representing 12 Wnt gene subfamilies in the Pacific white shrimp, Litopenaeus vannamei. Based on homolog annotations and phylogenetic analyses, we named these 12 Wnt genes as LvWnt1, LvWnt2, LvWnt4-11, LvWnt16, and LvWntA. All the corresponding LvWnt proteins shared a conserved Wnt1 domain and 22 conserved cysteine residues. LvWnt1 and LvWnt6 were adjacent in a scaffold in the shrimp genome. Furthermore, we performed expression analyses of LvWnt genes at different developmental stages, during the molting process, in different tissues and after different pathogenic infection. We showed that each LvWnt gene had a unique expression pattern at different developmental stages but only a few of them expressed in adult shrimp. All the investigated LvWnt genes were initially expressed at the gastrula or limb bud embryo stages. Among them, LvWnt8 was specifically high expressed only in early embryos. LvWntA and LvWnt5 displayed high and similar expression profiles during the molting process, and LvWnt6 and LvWnt16 were specifically expressed in the thoracic ganglion, ventral nerve, intestines and gill tissues, respectively. We also found the expression of LvWntA, LvWnt5, LvWnt6, LvWnt9, and LvWnt16 were varied in the different tissues after infected with Staphylococcus aureus, Vibrio parahaemolyticus and white spot syndrome virus (WSSV), which indicated that they might participate in immune response in L. vannamei. This study provided an insight into the repertoire of the Wnt gene structure and expression in shrimps, and furthermore, might promote the understanding of development, growth and immune response of shrimps and crustaceans.
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Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Penaeidae/genética , Penaeidae/inmunología , Proteínas Wnt/genética , Proteínas Wnt/inmunología , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/química , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/inmunología , Secuencia de Bases , Perfilación de la Expresión Génica , Familia de Multigenes/genética , Familia de Multigenes/inmunología , Filogenia , Alineación de Secuencia , Staphylococcus aureus/fisiología , Vibrio parahaemolyticus/fisiología , Virus del Síndrome de la Mancha Blanca 1/fisiología , Proteínas Wnt/químicaRESUMEN
Renal repair after injury is dependent on clonal expansion of proliferation-competent cells. In the human kidney, the expression of CD133 characterizes a population of resident scattered cells with resistance to damage and ability to proliferate. However, the biological function of the CD133 molecule is unknown. By RNA sequencing, we found that cells undergoing cisplatin damage lost the CD133 signature and acquired metanephric mesenchymal and regenerative genes such as SNAIL1, KLF4, SOX9, and WNT3. CD133 was reacquired in the recovery phase. In CD133-Kd cells, lack of CD133 limited cell proliferation after injury and was specifically correlated with deregulation of Wnt signaling and E-cadherin pathway. By immunoprecipitation, CD133 appeared to form a complex with E-cadherin and ß-catenin. In parallel, CD133-Kd cells showed lower ß-catenin levels in basal condition and after Wnt pathway activation and reduced TCF/LEF promoter activation in respect to CD133+ cells. Finally, the lack of CD133 impaired generation of nephrospheres while favoring senescence. These data indicate that CD133 may act as a permissive factor for ß-catenin signaling, preventing its degradation in the cytoplasm. Therefore, CD133 itself appears to play a functional role in renal tubular repair through maintenance of proliferative response and control of senescence. Stem Cells Translational Medicine 2018;7:283-294.
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Antígeno AC133/inmunología , Lesión Renal Aguda/inmunología , Riñón/inmunología , Proteínas Wnt/inmunología , Lesión Renal Aguda/genética , Línea Celular Tumoral , Proliferación Celular/genética , Proliferación Celular/fisiología , Técnicas de Silenciamiento del Gen , Humanos , Factor 4 Similar a Kruppel , TransfecciónRESUMEN
The Wnt signaling mediated by Wnt proteins that orchestrate and influence a myriad of cellular processes, such as cell proliferation, differentiation, tumorigenesis, apoptosis, and participation in immune defense during microbe infection. Wnt5b is one of the Wnt signaling molecules that initiate the cascade. In this study, we cloned and characterized a Wnt5b homolog from Litopenaeus vannamei designed as LvWnt5b. The full length of LvWnt5b transcript was 1726 bp with an 1107 bp open reading frame that encoded a 368 aa protein, which contained 24 discontinuous and highly conserved cysteine. Real-time quantitative PCR showed that the transcriptional level of LvWnt5b was down-regulated when infected with white spot syndrome virus (WSSV). Knock-down of LvWnt5b resulted in inhibition of the transcriptional level of WSSV gene ie1, indicating that LvWnt5b mediated signaling pathway may play an important role in defense against WSSV infection. When LvWnt5b was silenced, caspase3/7 activity in hemocytes was increased significantly, and the transcription of viral gene was decreased as well. Moreover, overexpression of LvWnt5b in HEK293T cells led to inhibition of caspase3/7 activity, which further proved the role of LvWnt5b in restraining apoptosis. The study showed that the shrimp may decrease the expression of LvWnt5b initiatively to act as an immune defense mechanism against WSSV infection via promoting apoptosis. It will be helpful for understanding the function of Wnt signaling pathway in virus invasion and host defense.
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Apoptosis/genética , Inmunidad Innata/genética , Penaeidae/genética , Penaeidae/inmunología , Virus del Síndrome de la Mancha Blanca 1/fisiología , Proteínas Wnt/genética , Proteínas Wnt/inmunología , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/química , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/inmunología , Secuencia de Bases , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/inmunología , Células HEK293 , Humanos , Filogenia , Transducción de Señal , Proteínas Wnt/químicaRESUMEN
The variation of macrophage functions suggests the involvement of multiple signalling pathways in fine tuning their differentiation. Macrophages that originate from monocytes in the blood migrate to tissue in response to homeostatic or 'danger' signals and undergo substantial morphological and functional modifications to meet the needs of the dominant signals in the microenvironment. Wnts are secreted glycoproteins that play a significant role in organ and cell differentiation, yet their impact on monocyte differentiation is not clear. In this study, we assessed the role of Wnt1 and Wnt7a on the differentiation of monocytes and the subsequent phenotype and function of monocyte-derived macrophages (MDMs). We show that Wnt7a decreased the expression of CD14, CD11b, CD163 and CD206, whereas Wnt1 had no effect. The Wnt7a effect on CD11b was also observed in the brain and spleen of Wnt7a-/- adult brain mouse tissue and in embryonic Wnt7a-/- tissue. Wnt7a reduced the phagocytic capacity of M-MDMs, decreased interleukin-10 (IL-10) and IL-12 secretion and increased IL-6 secretion. Collectively, these findings demonstrate that Wnt7a generates an MDM phenotype with both pro-inflammatory and alternative MDM cytokine profiles and reduced phagocytic capacity. As such, Wnt7a can have a significant impact on macrophage responses in health and disease.
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Citocinas/inmunología , Regulación de la Expresión Génica/inmunología , Macrófagos/inmunología , Monocitos/inmunología , Fagocitosis , Proteínas Wnt/inmunología , Animales , Antígenos CD/genética , Antígenos CD/inmunología , Encéfalo/inmunología , Citocinas/genética , Femenino , Humanos , Macrófagos/citología , Masculino , Ratones , Ratones Noqueados , Monocitos/citología , Bazo/inmunología , Proteínas Wnt/genéticaRESUMEN
Multiple cytokines produced by immune cells induce remodeling and aid in maintaining bone homeostasis through differentiation of bone-forming osteoblasts and bone-resorbing osteoclasts. Here, we investigate bone remodeling controlled by the tumor necrosis factor (TNF) superfamily cytokine LIGHT. LIGHT-deficient mice (Tnfsf14-/- ) exhibit spine deformity and reduced femoral cancellous bone mass associated with an increase in the osteoclast number and a slight decrease of osteoblasts compared with WT mice. The effect of LIGHT in bone cells can be direct or indirect, mediated by both the low expression of the anti-osteoclastogenic osteoprotegerin (OPG) in B and T cells and reduced levels of the pro-osteoblastogenic Wnt10b in CD8+ T cells in Tnfsf14-/- mice. LIGHT stimulation increases OPG levels in B, CD8+ T, and osteoblastic cells, as well as Wnt10b expression in CD8+ T cells. The high bone mass in Light and T- and B-cell-deficient mice (Rag- /Tnfsf14- ) supports the cooperative role of the immune system in bone homeostasis. These results implicate LIGHT as a potential target in bone disease. © 2017 American Society for Bone and Mineral Research.
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Remodelación Ósea/inmunología , Hueso Esponjoso/inmunología , Fémur/inmunología , Miembro 14 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/deficiencia , Animales , Linfocitos B/inmunología , Linfocitos B/patología , Remodelación Ósea/genética , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/patología , Hueso Esponjoso/patología , Fémur/fisiología , Ratones , Ratones Noqueados , Osteoblastos/inmunología , Osteoclastos/inmunología , Osteoclastos/patología , Osteoprotegerina/genética , Osteoprotegerina/inmunología , Miembro 14 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/inmunología , Proteínas Wnt/genética , Proteínas Wnt/inmunologíaRESUMEN
With regard to post-surgical pain, the incidence of chronic post-thoracotomy pain (CPTP) is second only to that caused by amputation and the underlying mechanism remains elusive. The emerging role of Wnts has been confirmed in the pathogenesis of neuropathic and inflammatory pain, both of which are known components of CPTP. We investigated whether Wnt3a and Wnt5a were involved in the development of CPTP, concerning their regulation of inflammatory responses in a previously established rat model. We observed up regulated protein levels of Wnt3a, Wnt5a, ß-catenin, and TLR4, along with activated astrocytes and pro-inflammatory cytokines, in both dorsal root ganglia and the spinal cord dorsal horn. Furthermore, intrathecal inhibition of Wnt5a but not Wnts relieved mechanical hyperalgesia, down regulated expression of TLR4, and inactivated astrocytes and pro-inflammatory cytokines. These results suggest Wnt5a, but not Wnts, contributes to the development of CPTP, possibly by regulating the inflammatory response.
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Dolor Crónico/inmunología , Dolor Crónico/prevención & control , Dolor Postoperatorio/inmunología , Dolor Postoperatorio/prevención & control , Toracotomía/efectos adversos , Proteína Wnt-5a/inmunología , Animales , Dolor Crónico/etiología , Factores Inmunológicos/inmunología , Masculino , Ratas , Ratas Sprague-Dawley , Proteínas Wnt/inmunologíaRESUMEN
A disturbed inflammatory response following myocardial infarction (MI) is associated with poor prognosis and increased tissue damage. Monocytes are key players in healing after MI, but little is known about the role of the cardiac niche in monocyte activation. This study investigated microenvironment-dependent changes in inflammatory monocytes after MI RNA sequencing analysis of murine Ly6Chigh monocytes on day 3 after MI revealed differential regulation depending on location. Notably, the local environment strongly impacted components of the WNT signaling cascade. Analysis of WNT modulators revealed a strong upregulation of WNT Inhibitory Factor 1 (WIF1) in cardiomyocytes-but not fibroblasts or endothelial cells-upon hypoxia. Compared to wild-type (WT) littermates, WIF1 knockout mice showed severe adverse remodeling marked by increased scar size and reduced ejection fraction 4 weeks after MI While FACS analysis on day 1 after MI revealed no differences in neutrophil numbers, the hearts of WIF1 knockouts contained significantly more inflammatory monocytes than hearts from WT animals. Next, we induced AAV-mediated cardiomyocyte-specific WIF1 overexpression, which attenuated the monocyte response and improved cardiac function after MI, as compared to control-AAV-treated animals. Finally, WIF1 overexpression in isolated cardiomyocytes limited the activation of non-canonical WNT signaling and led to reduced IL-1ß and IL-6 expression in monocytes/macrophages. Taken together, we investigated the cardiac microenvironment's interaction with recruited monocytes after MI and identified a novel mechanism of monocyte activation. The local initiation of non-canonical WNT signaling shifts the accumulating myeloid cells toward a pro-inflammatory state and impacts healing after myocardial infarction.
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Monocitos/inmunología , Infarto del Miocardio/inmunología , Miocardio/inmunología , Proteínas Wnt/inmunología , Proteínas Adaptadoras Transductoras de Señales , Animales , Proteínas de la Matriz Extracelular/genética , Proteínas de la Matriz Extracelular/inmunología , Corazón , Humanos , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/inmunología , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Interleucina-6/genética , Interleucina-6/inmunología , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Infarto del Miocardio/genética , Miocitos Cardíacos/inmunología , Transducción de Señal , Proteínas Wnt/genéticaRESUMEN
The etiology determines quality and extent of the immune response after udder infection (mastitis). Infections with Gram negative bacteria (e.g. Escherichia coli) will quickly elicit strong inflammation of the udder, fully activate its immune defence via pathogen receptor driven activation of IκB/NF-κB signaling. This often eradicates the pathogen. In contrast, Gram-positive bacteria (e.g. Staphylococcus aureus) will slowly elicit a much weaker inflammation and immune response, frequently resulting in chronic infections. However, it was unclear which immune regulatory pathways are specifically triggered by S. aureus causing this partial immune subversion. We therefore compared in first lactating cows the earliest (1-3 h) udder responses against infection with mastitis causing pathogens of either species. Global transcriptome profiling, bioinformatics analysis and experimental validation of key aspects revealed as S. aureus infection specific features the (i) failure to activating IκB/NF-κB signaling; (ii) activation of the wnt/ß-catenin cascade resulting in active suppression of NF-κB signaling and (iii) rearrangement of the actin-cytoskeleton through modulating Rho GTPase regulated pathways. This facilitates invasion of pathogens into host cells. Hence, S. aureus mastitis is characterized by eliciting unbalanced immune suppression rather than inflammation and invasion of S. aureus into the epithelial cells of the host causing sustained infection.
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Infecciones por Escherichia coli/inmunología , Escherichia coli/inmunología , Interacciones Huésped-Patógeno , Mastitis Bovina/inmunología , Infecciones Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Transcriptoma/inmunología , Citoesqueleto de Actina/inmunología , Citoesqueleto de Actina/patología , Citoesqueleto de Actina/ultraestructura , Animales , Bovinos , Células Epiteliales/inmunología , Células Epiteliales/patología , Células Epiteliales/ultraestructura , Escherichia coli/patogenicidad , Infecciones por Escherichia coli/genética , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/patología , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Inmunidad Innata , Glándulas Mamarias Animales/inmunología , Glándulas Mamarias Animales/microbiología , Glándulas Mamarias Animales/patología , Mastitis Bovina/genética , Mastitis Bovina/microbiología , Mastitis Bovina/patología , Inhibidor NF-kappaB alfa/genética , Inhibidor NF-kappaB alfa/inmunología , FN-kappa B/genética , FN-kappa B/inmunología , Transducción de Señal , Especificidad de la Especie , Infecciones Estafilocócicas/genética , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/patología , Staphylococcus aureus/patogenicidad , Proteínas Wnt/genética , Proteínas Wnt/inmunología , beta Catenina/genética , beta Catenina/inmunología , Proteínas de Unión al GTP rho/genética , Proteínas de Unión al GTP rho/inmunologíaRESUMEN
Members of the family of serine proteinase inhibitors, such as kallistatin, have been shown to inhibit canonical Wnt-TCF/LEF-ß-catenin signaling via their interactions with the Wnt co-receptor LRP6. Yet the effects of transgenic overexpression of anti-Wnt serpins on hematopoiesis and lymphopoiesis are not well known. We studied the effects of human kallistatin (SERPINA4) on Wnt reporter activity in various cell types throughout the hematopoietic system and associated impacts on circulating white blood cell profiles. Transgenic overexpression of kallistatin suppressed Wnt-TCF/LEF-ß-catenin signaling in bone marrow, as demonstrated using a Wnt reporter mouse. Further, kallistatin overexpression and treatment were associated with reduced Wnt-TCF/LEF-ß-catenin activity in CD34+ c-kit+ bone marrow cells and CD19+ B lymphocytes, with reduced levels of these populations in bone marrow and peripheral circulation, respectively. The presence of CD3+CD4+, CD3+CD8+, and CD3- NK1.1+ T lymphocytes were not significantly affected. Our data suggest that overexpression of kallistatin interferes with lymphopoiesis, ultimately impacting the level of circulating CD19+ B lymphocytes.
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Antígenos CD19/inmunología , Linfocitos B/inmunología , Linfopoyesis/inmunología , Serpinas/inmunología , Proteínas Wnt/inmunología , Vía de Señalización Wnt/inmunología , Animales , Antígenos CD/genética , Antígenos CD/inmunología , Antígenos CD/metabolismo , Antígenos CD19/genética , Antígenos CD19/metabolismo , Linfocitos B/citología , Linfocitos B/metabolismo , Pollos , Humanos , Ratones , Ratones Transgénicos , Serpinas/biosíntesis , Serpinas/genética , Linfocitos T/citología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Proteínas Wnt/antagonistas & inhibidores , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , Vía de Señalización Wnt/genéticaRESUMEN
Adiponectin secreted from adipocytes is an anti-diabetic and anti-atherogenic adipokine. Adiponectin level is known to fall significantly in obesity induced type 2 diabetes which worsen insulin sensitivity because of aberrant lipid management. However, underlying mechanism of adiponectin decrease in obese diabetic condition is yet unclear. We report here that lowering of plasma adiponectin coincided with the higher Fetuin A (FetA) level in high fat diet (HFD) induced obese diabetic mice. Knock down of FetA gene (FetAKD) elevated adiponectin level markedly in HFD mice, while reinforcement of FetA into FetAKDHFD mice reduced its level again. These results indicate FetA's involvement in the lowering of adiponectin level in obesity induced diabetic mice. Our findings to understand how FetA could affect adiponectin decrease demonstrated that FetA could enhance Wnt3a expression in the adipocyte of HFD mice. FetA addition to 3T3L1 adipocyte incubation elevated Wnt3a expression in a dose dependent manner. Overexpression of Wnt3a by FetA inhibited PPARγ and adiponectin. FetA failed to reduce PPARγ and adiponectin in Wnt3a gene knocked down 3T3L1` adipocytes. All these suggest that FetA mediate its inhibitory effect on adiponectin through Wnt3a-PPARγ pathway. Inhibition of adiponectin expression through FetA and Wnt3a significantly compromised with the activation of AMPK and its downstream signalling molecules which adversely affected lipid management causing loss of insulin sensitivity. Downregulation of adiponectin in inflamed adipocyte by FetA through the mediation of Wnt3a and PPARγ is a new report.
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Adipocitos/inmunología , Adiponectina/inmunología , Obesidad/inmunología , PPAR gamma/inmunología , Transducción de Señal , Proteínas Wnt/inmunología , alfa-2-Glicoproteína-HS/inmunología , Células 3T3-L1 , Animales , Células Cultivadas , Inflamación/inmunología , Resistencia a la Insulina , Lípidos/inmunología , Masculino , Ratones , Ratones Endogámicos BALB CRESUMEN
OBJECTIVES: Asthma is a chronic inflammatory, heterogeneous airway disease affecting millions of people around the world. Curcumin has been found to have anti-inflammatory and antifibrosis effects. Researchers reported that curcumin regulated Wnt/ß-catenin signaling in lots of cells. However, whether curcumin regulates the levels of Wnt/ß-Catenin signaling in lung tissues and DCs (dendritic cells) remains unclear. In this study, we assessed the effects of curcumin on DCs and asthma. METHODS: C57BL/6 mice immunized with OVA (ovalbumin) were challenged thrice with an aerosol of OVA every second day for 8 days. Dexamethasone or curcumin was administered intraperitoneally to OVA-immunized C57BL/6 mice on day 24 once a day for 9 days. Mice were analyzed for effects of curcumin on asthma, inflammatory cell infiltration and cytokine levels in lung tissue. DCs were isolated from mouse bone morrow. The surface markers CD40, CD86 and CD11c of DCs was detected by FACS (fluorescence activated cell sorting) and the function of DCs was detected by mixed lymphocyte reaction. The expression of GSK-3ß and ß-catenin was detected by Western Blot. RESULTS: Results showed that OVA increased the number of inflammatory factors in BALF (bronchoalveolar lavage fluid), elevated lung inflammation scores in mice. Curcumin dose-dependently reversed the alterations induced by OVA in the asthmatic mice. Curcumin activated Wnt/ß-catenin signaling pathway in DCs and asthmatic mouse lungs. CONCLUSIONS: Curcumin could influence the morphology and function of DCs, ease asthma symptom and inflammatory reaction through the activation of Wnt/ß-catenin signaling. These results provide new evidence new evidence for application of curcumin on asthma.
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Antiinflamatorios/farmacología , Asma/tratamiento farmacológico , Curcumina/farmacología , Neumonía/tratamiento farmacológico , Proteínas Wnt/efectos de los fármacos , beta Catenina/efectos de los fármacos , Animales , Asma/inmunología , Antígeno B7-2/biosíntesis , Biomarcadores , Líquido del Lavado Bronquioalveolar/citología , Antígeno CD11c/biosíntesis , Antígenos CD40/biosíntesis , Citocinas/efectos de los fármacos , Células Dendríticas/efectos de los fármacos , Dexametasona/farmacología , Modelos Animales de Enfermedad , Eosinófilos/efectos de los fármacos , Femenino , Mediadores de Inflamación/metabolismo , Ratones , Ratones Endogámicos C57BL , Neumonía/inmunología , Transducción de Señal , Proteínas Wnt/inmunología , beta Catenina/metabolismoRESUMEN
Wnt signaling plays a central role in tumor initiation and tumor progression. Mutations in Wnt pathway components, such as the tumor suppressor APC, lead to malignant transformation. While previous studies focused on Wnt-related changes in cancer cells, the impact of aberrant Wnt signaling on the tumor microenvironment is only beginning to emerge. In order to investigate the role of increased Wnt secretion on tumor growth and the microenvironment, we generated a novel germ cell tumor model by overexpressing the Wnt secretion factor Evi/Wls in mouse embryonic stem cells. Evi-overexpressing teratoma were characterized by enhanced tumor growth in supporting a tumor-promoting role of Wnt secretion. Interestingly, enhanced Evi expression correlated with impaired immune cell recruitment. Specifically, T- and B-cell infiltration was reduced in Evi-overexpressing teratomas, which was independent of teratoma size and differentiation. Our study suggests that Wnt secretion impairs immunosurveillance. Since immune cell infiltration has been shown to have prognostic value, the levels of secreted Wnt activity might impact the efficiency of cancer immunotherapy.