RESUMEN
Chronic infections, including Mycobacterium tuberculosis (Mtb)-caused tuberculosis (TB), can induce host immune exhaustion. However, the key checkpoint molecules involved in this process and the underlying regulatory mechanisms remain largely undefined, which impede the application of checkpoint-based immunotherapy in infectious diseases. Here, through adopting time-of-flight mass cytometry and transcriptional profiling to systematically analyze natural killer (NK) cell surface receptors, we identify leukocyte immunoglobulin like receptor B1 (LILRB1) as a critical checkpoint receptor that defines a TB-associated cell subset (LILRB1+ NK cells) and drives NK cell exhaustion in TB. Mechanistically, Mtb-infected macrophages display high expression of human leukocyte antigen-G (HLA-G), which upregulates and activates LILRB1 on NK cells to impair their functions by inhibiting mitogen-activated protein kinase (MAPK) signaling via tyrosine phosphatases SHP1/2. Furthermore, LILRB1 blockade restores NK cell-dependent anti-Mtb immunity in immuno-humanized mice. Thus, LILRB1-HLA-G axis constitutes a NK cell immune checkpoint in TB and serves as a promising immunotherapy target.
Asunto(s)
Antígenos HLA-G , Células Asesinas Naturales , Receptor Leucocitario Tipo Inmunoglobulina B1 , Mycobacterium tuberculosis , Tuberculosis , Receptor Leucocitario Tipo Inmunoglobulina B1/metabolismo , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Antígenos HLA-G/metabolismo , Antígenos HLA-G/genética , Antígenos HLA-G/inmunología , Humanos , Animales , Tuberculosis/inmunología , Tuberculosis/microbiología , Ratones , Mycobacterium tuberculosis/inmunología , Macrófagos/inmunología , Macrófagos/metabolismo , Receptores Inmunológicos/metabolismo , Receptores Inmunológicos/genética , Antígenos CDRESUMEN
Infectious diseases, such as Mycobacterium tuberculosis (Mtb)-caused tuberculosis (TB), remain a global threat exacerbated by increasing drug resistance. Host-directed therapy (HDT) is a promising strategy for infection treatment through targeting host immunity. However, the limited understanding of the function and regulatory mechanism of host factors involved in immune defense against infections has impeded HDT development. Here, we identify the ubiquitin ligase (E3) TRIM27 (tripartite motif-containing 27) as a host protective factor against Mtb by enhancing host macroautophagy/autophagy flux in an E3 ligase activity-independent manner. Mechanistically, upon Mtb infection, nuclear-localized TRIM27 increases and functions as a transcription activator of TFEB (transcription factor EB). Specifically, TRIM27 binds to the TFEB promoter and the TFEB transcription factor CREB1 (cAMP responsive element binding protein 1), thus enhancing CREB1-TFEB promoter binding affinity and promoting CREB1 transcription activity toward TFEB, eventually inducing autophagy-related gene expression as well as autophagy flux activation to clear the pathogen. Furthermore, TFEB activator 1 can rescue TRIM27 deficiency-caused decreased autophagy-related gene transcription and attenuated autophagy flux, and accordingly suppressed the intracellular survival of Mtb in cell and mouse models. Taken together, our data reveal that TRIM27 is a host defense factor against Mtb, and the TRIM27-CREB1-TFEB axis is a potential HDT-based TB target that can enhance host autophagy flux.Abbreviations: ATG5: autophagy related 5; BMDMs: bone marrow-derived macrophages; CFU: colony-forming unit; ChIP-seq: chromatin immunoprecipitation followed by sequencing; CREB1: cAMP responsive element binding protein 1; CTSB: cathepsin B; E3: ubiquitin ligase; EMSA: electrophoretic mobility shift assay; HC: healthy control; HDT: host-directed therapy; LAMP: lysosomal associated membrane protein; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MCOLN1: mucolipin TPR cation channel 1; Mtb: Mycobacterium tuberculosis; NLS: nuclear localization signal; PBMCs: peripheral blood mononuclear cells; PRKA/PKA: protein kinase cAMP-activated; qRT-PCR: quantitative real-time PCR; RFP: RET finger protein; TB: tuberculosis; TBK1: TANK binding kinase 1; TFEB: transcription factor EB; TRIM: tripartite motif; TSS: transcription start site; ULK1: unc-51 like autophagy activating kinase 1.
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Autofagia , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice , Mycobacterium tuberculosis , Tuberculosis , Autofagia/fisiología , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Animales , Tuberculosis/inmunología , Tuberculosis/microbiología , Tuberculosis/metabolismo , Humanos , Ratones , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Ratones Endogámicos C57BL , Proteínas de Motivos Tripartitos/metabolismo , Proteínas de Motivos Tripartitos/genética , Macrófagos/metabolismo , Macrófagos/microbiología , Células HEK293 , Regiones Promotoras Genéticas/genética , Proteínas de Unión al ADN , Proteínas NuclearesRESUMEN
Smartphones are a potentially powerful platform for scientific instruments. Here, we demonstrate speckle spectroscopy with smartphone-level hardware. This technique promises greater performance thresholds than traditional diffraction gratings. Light is injected into an optical fiber and the emergent speckle patterns are imaged by a reversed-lens smartphone camera. The smartphone then uses an algorithm, running on a mobile computing app, to determine, in less than one second, the (hitherto unknown) input spectrum. We reconstruct a variety of visible-wavelength (470-670 nm) single and multi-peaked spectra using a tunable source. The latter also include a metameric pair, i.e., two spectra that are different, yet represent colors that are indistinguishable to the human eye.
RESUMEN
The regulation of antiviral immunity is crucial in maintaining host immune homeostasis, a process that involves dynamic modulations of host organelles. The Golgi apparatus is increasingly perceived as a host organelle functioning as a critical platform for innate immunity, but the detailed mechanism by which it regulates antiviral immunity remains elusive. Here, we identify the Golgi-localized G protein-coupled receptor 108 (GPR108) as a regulator of type Ι interferon responses by targeting interferon regulatory factor 3 (IRF3). Mechanistically, GPR108 enhances the ubiquitin ligase Smad ubiquitylation regulatory factor 1 (Smurf1)-mediated K63-linked polyubiquitination of phosphorylated IRF3 for nuclear dot 10 protein 52 (NDP52)-dependent autophagic degradation, leading to suppression of antiviral immune responses against DNA or RNA viruses. Taken together, our study provides insight into the crosstalk between the Golgi apparatus and antiviral immunity via a dynamic and spatiotemporal regulation of GPR108-Smurf1 axis, thereby indicating a potential target for treating viral infection.
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Antivirales , Receptores Acoplados a Proteínas G , Ubiquitina-Proteína Ligasas , Antivirales/metabolismo , Aparato de Golgi/metabolismo , Inmunidad Innata , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitinación , Receptores Acoplados a Proteínas G/metabolismoRESUMEN
Tartrate-resistant acid phosphatase type 5 (TRAP5) is an enzyme that is highly expressed in activated macrophages and osteoclasts and plays important biological functions in mammalian immune defense systems. In the study, we investigated the functions of tartrate-resistant acid phosphatase type 5b from Oreochromis niloticus (OnTRAP5b). The OnTRAP5b gene has an open reading frame of 975 bp, which encodes a mature peptide consisting of 302 amino acids with a molecular weight of 33.448 kDa. The OnTRAP5b protein contains a metallophosphatase domain with metal binding and active sites. Phylogenetic analysis revealed that OnTRAP5b is clustered with TRAP5b of teleost fish and shares a high amino acid sequence similarity with other TRAP5b in teleost fish (61.73-98.15%). Tissues expression analysis showed that OnTRAP5b was most abundant in the liver and was also widely expressed in other tissues. Upon challenge with Streptococcus agalactiae and Aeromonas hydrophila in vivo and in vitro, the expression of OnTRAP5b was significantly up-regulated. Additionally, the purified recombinant OnTRAP5b ((r)OnTRAP5) protein exhibited optimal phosphatase activity at pH 5.0 and an ideal temperature of 50 °C. The Vmax, Km, and kcat of purified (r)OnTRAP5b were found to be 0.484 µmol × min-1 × mg-1, 2.112 mM, and 0.27 s-1 with respect to pNPP as a substrate, respectively. Its phosphatase activity was differentially affected by metal ions (K+, Na+, Mg2+, Ca2+, Mn2+, Cu2+, Zn2+, and Fe3+) and inhibitors (sodium tartrate, sodium fluoride, and EDTA). Furthermore, (r)OnTRAP5b was found to promote the expression of inflammatory-related genes in head kidney macrophages and induce reactive oxygen expression and phagocytosis. Moreover, OnTRAP5b overexpression and knockdown had a significant effect on bacterial proliferation in vivo. When taken together, our findings suggest that OnTRAP5b plays a significant role in the immune response against bacterial infection in Nile tilapia.
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Cíclidos , Enfermedades de los Peces , Infecciones Estreptocócicas , Animales , Cíclidos/genética , Cíclidos/microbiología , Inmunidad Innata/genética , Fosfatasa Ácida Tartratorresistente/genética , Fosfatasa Ácida Tartratorresistente/metabolismo , Filogenia , Proteínas de Peces/metabolismo , Infecciones Estreptocócicas/veterinaria , Streptococcus agalactiae/genética , Regulación de la Expresión Génica , Mamíferos/metabolismoRESUMEN
Ferroptosis is a lipid peroxidation-driven and iron-dependent programmed cell death involved in multiple physical processes and various diseases. Emerging evidence suggests that several pathogens manipulate ferroptosis for their pathogenicity and dissemination, but the underlying molecular mechanisms remain elusive. Here, we identify that protein tyrosine phosphatase A (PtpA), an effector secreted by tuberculosis (TB)-causing pathogen Mycobacterium tuberculosis (Mtb), triggers ferroptosis to promote Mtb pathogenicity and dissemination. Mechanistically, PtpA, through its Cys11 site, interacts with host RanGDP to enter host cell nucleus. Then, the nuclear PtpA enhances asymmetric dimethylation of histone H3 arginine 2 (H3R2me2a) via targeting protein arginine methyltransferase 6 (PRMT6), thus inhibiting glutathione peroxidase 4 (GPX4) expression, eventually inducing ferroptosis to promote Mtb pathogenicity and dissemination. Taken together, our findings provide insights into molecular mechanisms of pathogen-induced ferroptosis, indicating a potential TB treatment via blocking Mtb PtpA-host PRMT6 interface to target GPX4-dependent ferroptosis.
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Ferroptosis , Mycobacterium tuberculosis , Tuberculosis , Humanos , Virulencia , Tuberculosis/microbiología , Mycobacterium tuberculosis/metabolismo , Histonas/metabolismo , Proteínas Tirosina Fosfatasas/metabolismo , Peroxidación de Lípido , Proteínas Nucleares/metabolismo , Proteína-Arginina N-Metiltransferasas/metabolismoRESUMEN
Dysregulation of gut homeostasis is associated with irritable bowel syndrome (IBS), a chronic functional gastrointestinal disorder affecting approximately 11.2% of the global population. The poorly understood pathogenesis of IBS has impeded its treatment. Here, we report that the E3 ubiquitin ligase tripartite motif-containing 27 (TRIM27) is weakly expressed in IBS but highly expressed in inflammatory bowel disease (IBD), a frequent chronic organic gastrointestinal disorder. Accordingly, knockout of Trim27 in mice causes spontaneously occurring IBS-like symptoms, including increased visceral hyperalgesia and abnormal stool features, as observed in IBS patients. Mechanistically, TRIM27 stabilizes ß-catenin and thus activates Wnt/ß-catenin signaling to promote intestinal stem cell (ISC) self-renewal. Consistent with these findings, Trim27 deficiency disrupts organoid formation, which is rescued by reintroducing TRIM27 or ß-catenin. Furthermore, Wnt/ß-catenin signaling activator treatment ameliorates IBS symptoms by promoting ISC self-renewal. Taken together, these data indicate that TRIM27 is critical for maintaining gut homeostasis, suggesting that targeting the TRIM27/Wnt/ß-catenin axis could be a potential treatment strategy for IBS. Our study also indicates that TRIM27 might serve as a potential biomarker for differentiating IBS from IBD.
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Enfermedades Inflamatorias del Intestino , Síndrome del Colon Irritable , Animales , Ratones , beta Catenina , Autorrenovación de las Células , Proteínas de Unión al ADN , Homeostasis , Síndrome del Colon Irritable/patología , Ubiquitina-Proteína Ligasas , Intestinos/metabolismoRESUMEN
The inflammasome-mediated cleavage of gasdermin D (GSDMD) causes pyroptosis and inflammatory cytokine release to control pathogen infection, but how pathogens evade this immune response remains largely unexplored. Here we identify the known protein phosphatase PtpB from Mycobacterium tuberculosis as a phospholipid phosphatase inhibiting the host inflammasome-pyroptosis pathway. Mechanistically, PtpB dephosphorylated phosphatidylinositol-4-monophosphate and phosphatidylinositol-(4,5)-bisphosphate in host cell membrane, thus disrupting the membrane localization of the cleaved GSDMD to inhibit cytokine release and pyroptosis of macrophages. Notably, this phosphatase activity requires PtpB binding to ubiquitin. Disrupting phospholipid phosphatase activity or the ubiquitin-interacting motif of PtpB enhanced host GSDMD-dependent immune responses and reduced intracellular pathogen survival. Thus, pathogens inhibit pyroptosis and counteract host immunity by altering host membrane composition.
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Inflamasomas , Piroptosis , Citocinas/metabolismo , Inflamasomas/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de Unión a Fosfato/metabolismo , Fosfolípidos , Monoéster Fosfórico Hidrolasas/metabolismo , Ubiquitina/metabolismoRESUMEN
The eukaryotic-type protein kinase G (PknG), one of the eleven eukaryotic type serine-threonine protein kinase (STPK) in Mycobacterium tuberculosis (Mtb), is involved in mycobacterial survival within macrophages, presumably by suppressing phagosome and autophagosome maturation, which makes PknG an attractive drug target. However, the exact mechanism by which PknG inhibits pathogen clearance during mycobacterial infection remains largely unknown. Here, we show that PknG promotes macroautophagy/autophagy induction but inhibits autophagosome maturation, causing an overall effect of blocked autophagy flux and enhanced pathogen intracellular survival. PknG prevents the activation of AKT (AKT serine/threonine kinase) via competitively binding to its pleckstrin homology (PH) domain, leading to autophagy induction. Remarkably, PknG could also inhibit autophagosome maturation to block autophagy flux via targeting host small GTPase RAB14. Specifically, PknG directly interacts with RAB14 to block RAB14-GTP hydrolysis. Furthermore, PknG phosphorylates TBC1D4/AS160 (TBC1 domain family member 4) to suppress its GTPase-activating protein (GAP) activity toward RAB14. In macrophages and in vivo, PknG promotes Mtb intracellular survival through blocking autophagy flux, which is dependent on RAB14. Taken together, our data unveil a dual-functional bacterial effector that tightly regulates host autophagy flux to benefit pathogen intracellular survival.Abbreviations: AKT: AKT serine/threonine kinase; ATG5: autophagy related 5; BMDMs: bone marrow-derived macrophages; DTT: dithiothreitol; FBS: fetal calf serum; GAP: GTPase-activating protein; MOI: multiplicity of infection; Mtb: Mycobacterium tuberculosis; MTOR: mechanistic target of rapamycin kinase; OADC: oleic acid-albumin-dextrose-catalase; PC, phosphatidylcholine; PH: pleckstrin homology; PI3K: phosphoinositide 3-kinase; PknG: protein kinase G; PtdIns(3,4,5)P3: phosphatidylinositol(3,4,5)-trisphosphate; SQSTM1: sequestosome 1; STPK: serine-threonine protein kinase; TB: tuberculosis; TBC1D4: TBC1 domain family member 4; TPR: tetratricopeptide repeat; ULK1: unc-51 like autophagy activating kinase 1; WT: wild-type.
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Mycobacterium tuberculosis , Tuberculosis , Autofagia/fisiología , Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de GMP Cíclico/farmacología , Proteínas Activadoras de GTPasa/metabolismo , Humanos , Mycobacterium tuberculosis/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas , Proteínas Proto-Oncogénicas c-akt/metabolismo , Serina , Tuberculosis/microbiología , Proteínas de Unión al GTP rab/metabolismoRESUMEN
Tumour necrosis factor-α (TNF-α) is a multifunctional cytokine involved in immune system homeostasis, antimicrobial defence, regulation of apoptosis, cell proliferation and differentiation. Although the pro-inflammatory property of TNF-α has been made new progress, detailed research on host defence against bacterial infection and inducing apoptosis remains to be revealed in early vertebrates. Here, we reported the TNF-α homologue (ToTNF-α) from pufferfish (Takifugu obscurus). The open reading frame (ORF) of ToTNF-α was 753 bp, encoding a protein of 250 aa contained the TNF family signature and conserved cysteine residues. The mRNA expression of ToTNF-α had a wide range of tested tissues, with the highest expression in the skin. After Aeromonas hydrophila infection, the mRNA expression of ToTNF-α was significantly up-regulated both in vivo and in vitro experiments. After stimulation by recombinant protein of ToTNF-α ((r)ToTNF-α), the relative expressions of endogenous TNF-α, caspase 8, caspase 3, p53, and Bax inhibitor-1 in head kidney leucocytes were all notably up-regulated. These results showed that ToTNF-α might induce apoptosis depend on pro- and anti-apoptotic proteins at mRNA level. Moreover, flow cytometry analysis indicated that the (r)ToTNF-α can induce apoptosis of head kidney leucocytes. Taken together, these characteristics suggest that ToTNF-α can participate in immune response against A. hydrophila and induce apoptosis at mRNA and cellular level, which will help to understand the mechanism of apoptosis and immune response in teleost fish.
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Apoptosis , Enfermedades de los Peces/inmunología , Takifugu/inmunología , Factor de Necrosis Tumoral alfa/química , Aeromonas hydrophila/fisiología , Secuencia de Aminoácidos , Animales , Enfermedades de los Peces/microbiología , Proteínas de Peces/análisis , Regulación de la Expresión Génica , Infecciones por Bacterias Gramnegativas/veterinaria , ARN Mensajero , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Upon Mycobacterium tuberculosis (Mtb) infection, protein kinase G (PknG), a eukaryotic-type serine-threonine protein kinase (STPK), is secreted into host macrophages to promote intracellular survival of the pathogen. However, the mechanisms underlying this PknG-host interaction remain unclear. Here, we demonstrate that PknG serves both as a ubiquitin-activating enzyme (E1) and a ubiquitin ligase (E3) to trigger the ubiquitination and degradation of tumor necrosis factor receptor-associated factor 2 (TRAF2) and TGF-ß-activated kinase 1 (TAK1), thereby inhibiting the activation of NF-κB signaling and host innate responses. PknG promotes the attachment of ubiquitin (Ub) to the ubiquitin-conjugating enzyme (E2) UbcH7 via an isopeptide bond (UbcH7 K82-Ub), rather than the usual C86-Ub thiol-ester bond. PknG induces the discharge of Ub from UbcH7 by acting as an isopeptidase, before attaching Ub to its substrates. These results demonstrate that PknG acts as an unusual ubiquitinating enzyme to remove key components of the innate immunity system, thus providing a potential target for tuberculosis treatment.
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Mycobacterium tuberculosis , Proteínas Quinasas Dependientes de GMP Cíclico , Mycobacterium tuberculosis/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Ubiquitina/genética , Ubiquitina/metabolismo , Enzimas Ubiquitina-Conjugadoras/genética , Enzimas Ubiquitina-Conjugadoras/metabolismo , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , UbiquitinaciónRESUMEN
CD154, a member of the TNF superfamily, is a multifunctional molecule highly expressed in activated T cells, and plays important roles in T cell-dependent humoral immune response. In this study, CD154 of Nile tilapia (Oreochromis niloticus) was identified, and its functions in the T cell-dependent immune response were demonstrated. The open reading frame (ORF) of OnCD154 is 699 bp, encoding a protein of 232 amino acids with a 23 amino acid transmembrane region. Amino acid sequence of OnCD154 is highly homologous to that of other teleost fish, especially rainbow trout. Quantitative real-time PCR (qRT-PCR) demonstrated that mRNA of OnCD154 is highly expressed in immune organs, especially in spleen, thymus, gills, head kidney, etc. In addition, the anti-OnCD154 polyclonal antibody (anti-(r)OnCD154) was successfully prepared, and it can react with natural protein in head kidney leukocytes. Following two immunizations with keyhole limpet hemocyanin (KLH) in vivo, the significantly up-regulated expression level of OnCD154 mRNA appeared earlier (fifth day) and higher (42.9 folds) in the second challenge than the first on in head kidney. Further, after stimulation with KLH in vitro, the expressions of T cell-dependent immune response-related molecules (activated T cell specific surface molecules CD3ε and CD154) and B cell differentiation-related molecules (Blimp1 and sIgM) and CD40 were significantly up-regulated in head kidney leukocytes. Moreover, the up-regulated expressions of these molecules were blocked with the treatment of anti-(r)OnCD154 antibody. Taken together, these results indicate that OnCD154 might get involved in T cell-dependent immune response, and provide a new insight into the humoral immune response of teleost fish.
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Ligando de CD40/genética , Ligando de CD40/inmunología , Cíclidos/genética , Cíclidos/inmunología , Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Humoral/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Ligando de CD40/química , Femenino , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Filogenia , Alineación de Secuencia/veterinaria , TranscriptomaRESUMEN
Previous reports have suggested a link between pulmonary tuberculosis (TB), which is caused by Mycobacterium tuberculosis (Mtb), and the development of lung adenocarcinoma (LUAD) and sarcoidosis. Furthermore, these lung diseases share certain clinical similarities that can challenge differential diagnosis in some cases. Here, through comparison of lung transcriptome-derived molecular signatures of TB, LUAD and sarcoidosis patients, we identify certain shared disease-related expression patterns. We also demonstrate that MKI67, an over-expressed gene shared by TB and LUAD, is a key mediator in Mtb-promoted tumor cell proliferation, migration, and invasion. Moreover, we reveal a distinct ossification-related TB lung signature, which may be associated with the activation of the BMP/SMAD/RUNX2 pathway in Mtb-infected macrophages that can restrain mycobacterial survival and promote osteogenic differentiation of mesenchymal stem cells. Taken together, these findings provide novel pathogenic links and potential molecular markers for better understanding and differential diagnosis of pulmonary TB, LUAD and sarcoidosis.
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Adenocarcinoma/genética , Neoplasias Pulmonares/genética , Sarcoidosis/genética , Transcriptoma/genética , Tuberculosis Pulmonar/genética , Adenocarcinoma/diagnóstico , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Biomarcadores , Diagnóstico Diferencial , Humanos , Pulmón/metabolismo , Pulmón/patología , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Sarcoidosis/diagnóstico , Sarcoidosis/metabolismo , Sarcoidosis/patología , Transducción de Señal/genética , Tuberculosis Pulmonar/diagnóstico , Tuberculosis Pulmonar/metabolismo , Tuberculosis Pulmonar/patologíaRESUMEN
Siglec-1, one of the sialic acid-binding immunoglobulin-type lectins, is closely related to the recognition of host-pathogen and cell-cell interactions in the adaptive and innate immune systems. In this communication, a Siglec-1-like gene (OnSiglec-1-like) from Nile tilapia (Oreochromis niloticus) was analyzed. Relative expression revealed that the OnSiglec-1-like was expressed in all tested tissues, and the highest expression was found in the anterior kidney. Upon Streptococcus agalactiae (S. agalactiae) infection, the expression of OnSiglec-1-like was up-regulated in anterior kidney and spleen significantly in vivo. Additionally, the same phenomenon was observed in anterior kidney leukocytes upon LPS and S. agalactiae challenges as well in vitro. Western-blotting and ELISA analyses revealed that recombinant OnSiglec-1-like protein possessed high binding activity to LTA, LPS and S. agalactiae. Further, the recombinant OnSiglec-1-like was able to agglutinate S. agalactiae. Moreover, with the digestion of specific sialidase, the phagocytic ability of macrophages to S. agalactiae was greatly enhanced. Taken together, these results indicated that the Siglec-1-like possesses conserved functions of agglutination and promotion of macrophage phagocytic activity in Nile tilapia.
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Cíclidos/genética , Cíclidos/inmunología , Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Lectina 1 Similar a Ig de Unión al Ácido Siálico/genética , Lectina 1 Similar a Ig de Unión al Ácido Siálico/inmunología , Inmunidad Adaptativa/genética , Aglutinación/inmunología , Animales , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Inmunidad Innata/genética , Macrófagos/inmunología , Fagocitosis/inmunología , Infecciones Estreptocócicas/inmunología , Infecciones Estreptocócicas/veterinaria , Streptococcus agalactiae/fisiologíaRESUMEN
Interleukin 6 (IL-6) is a pleiotropic cytokine that exerts its biological functions through interaction with its receptor system consisting of a ligand-specific IL-6 receptor (IL-6R) and a common signal-transducing receptor (gp130). In this study, OnIL-6R and Ongp130 genes from Nile tilapia (Oreochromis niloticus) were identified, and their roles in bacterial or viral infection and in regulation of inflammatory response involved in IL-6 were investigated. The open reading frames (ORFs) of OnIL-6R and Ongp130 are 2019 bp and 2679 bp, encoding 672 and 892 amino acids, respectively. Domain analysis of the deduced amino acid sequences of OnIL-6R and Ongp130 showed that both of them contained a conserved Ig-like domain, FNIII domains, and a WSXWS motif. The transcripts of OnIL-6R and Ongp130 were widely expressed in all examined tissues. Following in vivo challenges with Streptococcus agalactia, Poly I: C and lipopolysaccharide (LPS), the mRNAs of OnIL-6R and Ongp130 were notably induced in liver, head kidney and spleen. The transcriptional up-regulations of OnIL-6R and Ongp130 were also detected in Nile tilapia monocytes/macrophages and lymphocytes after in vitro stimulations with S. agalactiae, Poly I: C and LPS. Besides, increasing mRNA levels of the inflammation-related cytokines (IL-1ß, TNF-α, IL-6, IL-10, and MIF) induced by recombinant OnIL-6 could be further enhanced by co-treatment with recombinant soluble OnIL-6R in lymphocytes. Furthermore, recombinant soluble Ongp130 suppressed the induction of expression of these cytokines in lymphocytes when co-stimulated with (r)OnIL-6 and (r)sOnIL-6R. Taken together, these results indicated that OnIL-6R and Ongp130 were likely involved in the resistance to bacterial or viral infection in Nile tilapia. Moreover, soluble OnIL-6R and soluble Ongp130 have an agonistic effect or antagonistic effect in the inflammation response involved in OnIL-6.
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Cíclidos/inmunología , Receptor gp130 de Citocinas/genética , Proteínas de Peces/genética , Receptores de Interleucina-6/genética , Infecciones Estreptocócicas/inmunología , Streptococcus agalactiae/fisiología , Virosis/inmunología , Animales , Clonación Molecular , Receptor gp130 de Citocinas/metabolismo , Citocinas/metabolismo , Resistencia a la Enfermedad , Proteínas de Peces/metabolismo , Inmunidad Innata , Mediadores de Inflamación/metabolismo , Poli I-C/inmunología , Receptores de Interleucina-6/metabolismo , Regulación hacia ArribaRESUMEN
Microwave-assisted pyrolysis of waste suffers from the problem that the waste generally has low microwave absorptivity thereby resulting in low heating rate and low pyrolysis temperature. In this case, fast microwave-assisted pyrolysis is proposed and developed to help the pyrolysis of waste. This study describes two methods that can be used to realize fast microwave-assisted pyrolysis of waste: (1) premixed method (wastes are mixed with microwave absorbent) and (2) non-premixed method (wastes are poured onto the heated microwave absorbent bed). Then, biofuels (bio-oil, bio-gas, and bio-char) produced from fast microwave-assisted pyrolysis of wastes are reviewed. The review results show that the yields of bio-oil, bio-gas, and bio-char obtained from fast microwave-assisted pyrolysis of wastes varied significantly in the ranges of 2-96 wt%, 2.4-86.8 wt%, and 0.3-83.2 wt%, respectively. Although the present research focused mainly on the premixed method, non-premixed/continuous fast microwave-assisted pyrolysis is still promising and challenging.
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Biocombustibles , Microondas , Calefacción , Calor , PirólisisRESUMEN
Complement component 1q (C1q), together with C1r and C1s to form C1, recognize and bind immune complex to initiate the classical complement pathway. In this study, C1q subunit molecules (XlC1qA, XlC1qB, XlC1qC) were cloned and analyzed from Xenopus laevis (X. laevis). The open reading frame (ORF) of XlC1qA is 819 bp of nucleotide sequence encoding 272 amino acids, the ORF of XlC1qB is 711 bp encoding 236 aa, and the XlC1qC is consists of 732 bp encoding 243 aa. The deduced amino acid sequences contain a collagen-like region (CLR), Gly-X-Y repeats in the N-terminus and a C1q family domain at the C-terminus. Phylogenetic analysis revealed that the XlC1qs are clustered with the amphibian clade. Expression analysis indicated that the XlC1qs exhibited constitutive expression in all examined tissues, with the highest expression in liver. Additionally, XlC1q could interact with heat-aggregated mouse IgG and IgM, Xenopus IgM and Nile tilapia IgM, respectively, indicating the functional conservation of XlC1q binding to immunoglobulins. Further, XlC1qs can inhibit C1q-dependent hemolysis of sensitized sheep red blood cells with concentration-dependent manner. These data collectively suggest that the function of C1qs in X. laevis may be conserved in interaction with immunoglobulins, as that of mammals and teleosts.
Asunto(s)
Complemento C1q/inmunología , Xenopus laevis/inmunología , Animales , Inmunoglobulina G/inmunología , Inmunoglobulina M/inmunologíaRESUMEN
In this study, purification of polysaccharide ulvan by anion exchange chromatography was prepared, and the major polysaccharide fraction (FU) was collected at 1.0â¯M NaCl elute by anion exchange chromatography, then high sulfate content purified ulvan (HFU) was prepared with sulfur trioxide/N,N-dimethylformamide (SO3-DMF) in formamide. The antioxidant activity and the antihyperlipidemic activity of HFU in mice were determined. The results showed that treatment with HFU could improve the antioxidant and antihyperlipidemic activity. Compared with the hyperlipidemic group, the antihyperlipidemic activity of HFU (125â¯mg/kg) was the strongest, TC concentrations were significantly decreased by 26.7% (Pâ¯<â¯.01), significantly reduced LDL-C (32.6%, Pâ¯<â¯.01), significantly increased HDL-C (19.6%, Pâ¯<â¯.01), and HFU-treated group (250â¯mg/kg) exhibited optimal effects on TG (29.0%, Pâ¯<â¯.01); the HFU groups at the doses of 125â¯mg/kg could significantly decrease the MDA (29.9%, Pâ¯<â¯.01); the HFU groups at the doses of 500â¯mg/kg could increase the activities of GSH-Px obviously (19.9%, Pâ¯<â¯.01).
Asunto(s)
Antioxidantes/farmacología , Hipolipemiantes/farmacología , Extractos Vegetales/farmacología , Polisacáridos/química , Sulfatos/química , Ulva/química , Animales , Antioxidantes/química , Peso Corporal/efectos de los fármacos , Modelos Animales de Enfermedad , Radical Hidroxilo/análisis , Hipolipemiantes/química , Masculino , Ratones , Peso Molecular , Monosacáridos/aislamiento & purificación , Extractos Vegetales/química , Superóxidos/análisisRESUMEN
CD38 is a multifunctional cell surface molecule that plays a crucial role in B cell activation, differentiation, and maturation in mammals with an increased expression in B cell maturation. In this study, a CD38-like molecule (OnCD38) was cloned and identified from Nile tilapia (Oreochromis niloticus), and its functional characterization was investigated. The open reading frame of OnCD38 is 828 bp of the nucleotide sequence, encoding a polypeptide of 275â¯amino acids. The deduced amino acid sequence of OnCD38 is highly homologous to other teleost fish and similar to mammals, containing extracellular, intracellular and transmembrane regions. Subcellular localization studies revealed that OnCD38 molecules were presented on the surface of B cells. Three healthy tilapia were used in each experimental group and control group. Following keyhole limpet hemocyanin (KLH) challenge in vivo, the mRNA expression of OnCD38 was significantly up-regulated in peripheral blood, spleen, and head kidney, with an earlier up-regulation in the second challenge than the first one. The up-regulation of OnCD38 expression was also detected in head kidney leukocytes after stimulation with LPS, recombinant HomoIL-10 ((r)HomoIL-10), (r)OnIL-10, and LPS plus (r)OnIL-10 in vitro. Furthermore, the OnCD38 expression increased with the differentiation of B cells, reaching a high level (10.1 fold higher than resting mature B cells) at the plasma-like B cells. Taken together, in this study, these results indicate that the OnCD38 is likely involved in the T cell-dependent response and plays roles in B cell differentiation in Nile tilapia.
Asunto(s)
ADP-Ribosil Ciclasa 1/inmunología , Linfocitos B/inmunología , Cíclidos/inmunología , Proteínas de Peces/inmunología , Linfocitos T/inmunología , Animales , Diferenciación Celular/inmunología , Activación de Linfocitos/inmunologíaRESUMEN
Ulvan was the polysaccharide (U) from a large edible green algae Ulva pertusa. In this study, phosphorylated ulvan (PU) was prepared by the sodium trimetaphosphate -sodium tripolyphosphate method. Antioxidant and antihyperlipidemic effects of U and PU were investigated employing in vivo systems. The PU was confirmed by IR, 31P NMR and 13C NMR spectra. And in addition, we found that the PU3 group at the dose of 500 mg/kg showed stronger antioxidant activity. Compared with hyperlipidemia group, it significantly increased GSH-Px (34.29%; P < 0.01), SOD (20.04%; P < 0.01) and CAT (37.49%; P < 0.01). Treatment of hyperlipidemia mice with PU resulted in a significant decrease in TC, TG and LDL-C, and significant increase in HDL-C. The PU3 significantly increased HDL-C (33.70%; P < 0.01), decreased LDL-C (52.73%; P < 0.01) and TG (33.58%; P < 0.01) compared with hyperlipidemia group. The result showed that phosphorylation could improve hypolipidaemic and antioxidant activities in vivo. PU may be used as a drug for hyperlipidaemia treatment.