RESUMEN
A subpopulation of deeply quiescent, so-called dormant hematopoietic stem cells (dHSCs) resides at the top of the hematopoietic hierarchy and serves as a reserve pool for HSCs. The state of dormancy protects the HSC pool from exhaustion throughout life; however, excessive dormancy may prevent an efficient response to hematological stresses. Despite the significance of dHSCs, the mechanisms maintaining their dormancy remain elusive. Here, we identify CD38 as a novel and broadly applicable surface marker for the enrichment of murine dHSCs. We demonstrate that cyclic adenosine diphosphate ribose (cADPR), the product of CD38 cyclase activity, regulates the expression of the transcription factor c-Fos by increasing the release of Ca2+ from the endoplasmic reticulum (ER). Subsequently, we uncover that c-Fos induces the expression of the cell cycle inhibitor p57Kip2 to drive HSC dormancy. Moreover, we found that CD38 ecto-enzymatic activity at the neighboring CD38-positive cells can promote human HSC quiescence. Together, CD38/cADPR/Ca2+/c-Fos/p57Kip2 axis maintains HSC dormancy. Pharmacological manipulations of this pathway can provide new strategies to improve the success of stem cell transplantation and blood regeneration after injury or disease.
Asunto(s)
ADP-Ribosil Ciclasa 1 , ADP-Ribosa Cíclica , Animales , Humanos , Ratones , Calcio/metabolismo , ADP-Ribosa Cíclica/metabolismo , Células Madre Hematopoyéticas , ADP-Ribosil Ciclasa 1/metabolismo , Inhibidor p57 de las Quinasas Dependientes de la Ciclina/metabolismoRESUMEN
HyperSpectral (HS) images have been successfully used for brain tumor boundary detection during resection operations. Nowadays, these classification maps coexist with other technologies such as MRI or IOUS that improve a neurosurgeon's action, with their incorporation being a neurosurgeon's task. The project in which this work is framed generates an unified and more accurate 3D immersive model using HS, MRI, and IOUS information. To do so, the HS images need to include 3D information and it needs to be generated in real-time operating room conditions, around a few seconds. This work presents Graph cuts Reference depth estimation in GPU (GoRG), a GPU-accelerated multiview depth estimation tool for HS images also able to process YUV images in less than 5.5 s on average. Compared to a high-quality SoA algorithm, MPEG DERS, GoRG YUV obtain quality losses of -0.93 dB, -0.6 dB, and -1.96% for WS-PSNR, IV-PSNR, and VMAF, respectively, using a video synthesis processing chain. For HS test images, GoRG obtains an average RMSE of 7.5 cm, with most of its errors in the background, needing around 850 ms to process one frame and view. These results demonstrate the feasibility of using GoRG during a tumor resection operation.
Asunto(s)
Algoritmos , Neoplasias Encefálicas , Encéfalo , Humanos , Imagen por Resonancia MagnéticaRESUMEN
Transferrin purification from mice serum samples by immunoaffinity chromatography (IAC) was optimized in order to study the possible modifications occurring in its glycans in collagen-induced arthritis (CIA) samples. SDS-PAGE and nanoLC-MS/MS were used to monitor the IAC purification performance. Afterward, a relative quantification of mouse transferrin (mTf) glycan isomers using [12C6]/[13C6]-aniline was used to unequivocally detect alterations in the glycan profile of CIA mice. In addition, multivariate data analysis was applied to identify the most meaningful glycan isomers for the discrimination between control and pathological samples. Partial least-squares discriminant analysis (PLS-DA) revealed that five out of fifteen mTf glycan isomers could be potential biomarkers of CIA, most of them corresponding to highly sialylated structures (H6N5S3_2, H6N5S3_3, and H5N4S3_2). Moreover, some of these glycan isomers also seemed to be related with the progression of CIA, especially H6N5S2 and H6N5S3_2, as their overexpression increased with the clinical score of the pathology. Hence, the established methodology not only provides valuable information to find glycan-based biomarkers of CIA, but also leaves the door open to evaluate, in the future, glycosylation changes of many other inflammatory diseases, in which transferrin has been described to be altered.
Asunto(s)
Artritis Experimental , Transferrina , Animales , Glicosilación , Ratones , Polisacáridos , Espectrometría de Masas en TándemRESUMEN
Previous work from our group has shown that Cd38-/- mice develop a milder pristane-induced lupus disease than WT or Art2-/- counterparts, demonstrating a new role for CD38 in promoting aberrant inflammation and lupus-like autoimmunity via a Transient Receptor Potential Melastatin 2 (TRPM2)-dependent apoptosis-driven mechanism. In this study we asked whether CD38 may play a role in the expression and function of regulatory B cells (IL-10-producing B cells or B10 cells). In pristane-treated mice the frequency of spleen CD19âºCD1dhiCD5⺠B cells, which are highly enriched in B10 cells, was significantly increased in Cd38-/- splenocytes compared to WT, while the frequency of peritoneal plasmacytoid dendritic cells (pDCs), which are major type I Interferon (IFN) producers, was greatly diminished. The low proportion of pDCs correlated with lower amounts of IFN-α in the peritoneal lavage fluids of the Cd38-/- mice than of WT and Art2-/- mice. Functional ex vivo assays showed increased frequencies of IL-10-producing B cells in Cd38-/- splenocytes than in WT upon stimulation with an agonist anti-CD40 mAb. Overall these results strongly suggest that Cd38-/- mice are better suited than WT mice to generate and expand regulatory B10 cells following the appropriate stimulation.
Asunto(s)
ADP-Ribosil Ciclasa 1/inmunología , Linfocitos B Reguladores/inmunología , Lupus Eritematoso Sistémico/inmunología , Glicoproteínas de Membrana/inmunología , ADP-Ribosil Ciclasa 1/genética , Animales , Autoinmunidad , Linfocitos B Reguladores/patología , Células Dendríticas/inmunología , Células Dendríticas/patología , Modelos Animales de Enfermedad , Eliminación de Gen , Interferón Tipo I/inmunología , Interleucina-10/inmunología , Lupus Eritematoso Sistémico/patología , Glicoproteínas de Membrana/genética , Ratones , Ratones Endogámicos C57BLRESUMEN
Leishmaniasis is important as a cause of hemophagocytic lymphohistiocytosis (HLH) and must be considered and excluded in patients with HLH because it can cause severe or even fatal complications. When HLH is present, there is a deficient downregulation of the immune response, leading to an uncontrolled inflammation. We report a case of visceral leishmaniasis-HLH where the therapy with tocilizumab, targeting interleukin 6, help to regulate the immune response for the infection of Leishmania.
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Anticuerpos Monoclonales Humanizados/administración & dosificación , Leishmaniasis Visceral/complicaciones , Linfohistiocitosis Hemofagocítica/tratamiento farmacológico , Adulto , Anticuerpos Monoclonales Humanizados/farmacología , Anticuerpos Monoclonales Humanizados/uso terapéutico , Femenino , Humanos , Leishmaniasis Visceral/tratamiento farmacológico , Leishmaniasis Visceral/inmunología , Linfohistiocitosis Hemofagocítica/etiología , Linfohistiocitosis Hemofagocítica/inmunología , Receptores de Interleucina-6/antagonistas & inhibidores , Receptores de Interleucina-6/inmunologíaRESUMEN
Collagen-type-II-induced arthritis (CIA) is an autoimmune disease, which involves a complex host systemic response including inflammatory and autoimmune reactions. CIA is milder in CD38(-/-) than in wild-type (WT) mice. ProteoMiner-equalized serum samples were subjected to 2D-DiGE and MS-MALDI-TOF/TOF analyses to identify proteins that changed in their relative abundances in CD38(-/-) versus WT mice either with arthritis (CIA(+) ), with no arthritis (CIA(-) ), or with inflammation (complete Freund's adjuvant (CFA)-treated mice). Multivariate analyses revealed that a multiprotein signature (n = 28) was able to discriminate CIA(+) from CIA(-) mice, and WT from CD38(-/-) mice within each condition. Likewise, a distinct multiprotein signature (n = 16) was identified which differentiated CIA(+) CD38(-/-) mice from CIA(+) WT mice, and lastly, a third multiprotein signature (n = 18) indicated that CD38(-/-) and WT mice could be segregated in response to CFA treatment. Further analyses showed that the discriminative power to distinguish these groups was reached at protein species level and not at the protein level. Hence, the need to identify and quantify proteins at protein species level to better correlate proteome changes with disease processes. It is crucial for plasma proteomics at the low-abundance protein species level to apply the ProteoMiner enrichment. All MS data have been deposited in the ProteomeXchange with identifiers PXD001788, PXD001799 and PXD002071 (http://proteomecentral.proteomexchange.org/dataset/PXD001788, http://proteomecentral.proteomexchange.org/dataset/PXD001799 and http://proteomecentral.proteomexchange.org/dataset/PXD002071).
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Artritis Experimental/sangre , Inflamación/sangre , Proteoma/análisis , ADP-Ribosil Ciclasa 1/genética , Animales , Artritis Experimental/complicaciones , Artritis Experimental/fisiopatología , Adyuvante de Freund , Inflamación/inducido químicamente , Masculino , Glicoproteínas de Membrana/genética , Ratones , Ratones Noqueados , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Electroforesis Bidimensional Diferencial en GelRESUMEN
PURPOSE: Magnetic resonance imaging (MRI) is a common technique in image-guided neurosurgery (IGN). Recent research explores the integration of methods like ultrasound and tomography, among others, with hyperspectral (HS) imaging gaining attention due to its non-invasive real-time tissue classification capabilities. The main challenge is the registration process, often requiring manual intervention. This work introduces an automatic, markerless method for aligning HS images with MRI. METHODS: This work presents a multimodal system that combines RGB-Depth (RGBD) and HS cameras. The RGBD camera captures the patient's facial geometry, which is used for registration with the preoperative MR through ICP. Once MR-depth registration is complete, the integration of HS data is achieved using a calibrated homography transformation. The incorporation of external tracking with a novel calibration method allows camera mobility from the registration position to the craniotomy area. This methodology streamlines the fusion of RGBD, HS and MR images within the craniotomy area. RESULTS: Using the described system and an anthropomorphic phantom head, the system has been characterised by registering the patient's face in 25 positions and 5 positions resulted in a fiducial registration error of 1.88 ± 0.19 mm and a target registration error of 4.07 ± 1.28 mm, respectively. CONCLUSIONS: This work proposes a new methodology to automatically register MR and HS information with a sufficient accuracy. It can support the neurosurgeons to guide the diagnosis using multimodal data over an augmented reality representation. However, in its preliminary prototype stage, this system exhibits significant promise, driven by its cost-effectiveness and user-friendly design.
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Imagen por Resonancia Magnética , Procedimientos Neuroquirúrgicos , Fantasmas de Imagen , Humanos , Imagen por Resonancia Magnética/métodos , Procedimientos Neuroquirúrgicos/métodos , Procedimientos Neuroquirúrgicos/instrumentación , Cirugía Asistida por Computador/métodos , Imágenes Hiperespectrales/métodos , Imagen Multimodal/métodos , Imagen Multimodal/instrumentaciónRESUMEN
CD38 is a multifunctional protein possessing ADP-ribosyl cyclase activity responsible for both the synthesis and the degradation of several Ca(2+)-mobilizing second messengers. In mammals, CD38 also functions as a receptor. In this study CD38 expression in CD4(+), CD8(+), or CD25(+) T cells was significantly higher in systemic lupus erythematosus (SLE) patients than in Normal controls. Increased CD38 expression in SLE T cells correlated with plasma levels of Th2 (IL-4, IL-10, IL-13) and Th1 (IL-1ß, IL-12, IFN-γ, TNF-α) cytokines, and was more prevalent in clinically active SLE patients than in Normal controls. In contrast, elevated anti-CD38 IgG autoantibodies were more frequent in clinically quiescent SLE patients (SLEDAI=0) than in Normal controls, and correlated with moderate increased plasma levels of IL-10 and IFN-γ. However, clinically active SLE patients were mainly discriminated from quiescent SLE patients by increased levels of IL-10 and anti-dsDNA antibodies, with odds ratios (ORs) of 3.7 and 4.8, respectively. Increased frequency of anti-CD38 autoantibodies showed an inverse relationship with clinical activity (OR=0.43), and in particular with the frequency of anti-dsDNA autoantibodies (OR=0.21). Increased cell death occurred in CD38(+) Jurkat T cells treated with anti-CD38(+) SLE plasmas, and not in these cells treated with anti-CD38(-) SLE plasmas, or Normal plasmas. This effect did not occur in CD38-negative Jurkat T cells, suggesting that it could be attributed to anti-CD38 autoantibodies. These results support the hypothesis that anti-CD38 IgG autoantibodies or their associated plasma factors may dampen immune activation by affecting the viability of CD38(+) effector T cells and may provide protection from certain clinical SLE features.
Asunto(s)
ADP-Ribosil Ciclasa 1/inmunología , Autoanticuerpos/sangre , Inmunoglobulina G/inmunología , Lupus Eritematoso Sistémico/inmunología , Lupus Eritematoso Sistémico/metabolismo , Subgrupos de Linfocitos T/inmunología , ADP-Ribosil Ciclasa 1/biosíntesis , Anticuerpos Antinucleares/sangre , Anticuerpos Antinucleares/inmunología , Autoanticuerpos/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Línea Celular Tumoral , Citocinas/biosíntesis , Citocinas/sangre , Femenino , Humanos , Inmunoglobulina G/sangre , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Células Jurkat , Lupus Eritematoso Sistémico/sangre , Activación de Linfocitos , Recuento de Linfocitos , Masculino , Fenotipo , Subgrupos de Linfocitos T/metabolismoRESUMEN
BACKGROUND: Chronic inflammation plays an important role in the development of cardiovascular risk factors. Although the prevalence of comorbidities and cardiovascular events has been described in patients with psoriasis, few studies have examined subclinical atherosclerosis in psoriasis patients. OBJECTIVE: Our objective was to investigate the prevalence of atheroma plaques in patients with severe psoriasis compared with control subjects and to analyze the association with metabolic syndrome, homocysteine levels and inflammatory parameters. PATIENTS AND METHODS: This case-control study included 133 patients, 72 with psoriasis and 61 controls consecutively admitted to the outpatient clinic in Dermatology Departments (Granada, Spain.) RESULTS: Carotid atheroma plaques were observed in 34.7% of the psoriatic patients versus 8.2% of the controls (p=0.001) and metabolic syndrome was diagnosed in 40.3% of the psoriatic patients versus 13.1% of the controls (p<0.001). Significantly higher mean values of insulin, aldosterone, homocysteine and acute phase parameters (fibrinogen, D-dimer, C reactive protein and erythrocyte sedimentation rate) were found in psoriatic patients. Binary logistic regression showed a strong association between psoriasis and atheroma plaque and metabolic syndrome after controlling for confounding variables. LIMITATIONS: The absence of longitudinal quantification of metabolic syndrome parameters and intima-media thickness in psoriatic patients. CONCLUSION: The chronic inflammation and hyperhomocysteinemia found in psoriatic patients may explain the association with atheroma plaque and metabolic syndrome. Cardiovascular screening by metabolic syndrome criteria assessment and carotid ultrasound in psoriasis may be useful to detect individuals at risk and start preventive treatment against the development of cardiovascular disease.
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Síndrome Metabólico/epidemiología , Placa Aterosclerótica/epidemiología , Psoriasis/epidemiología , Proteínas de Fase Aguda/análisis , Adulto , Enfermedades de las Arterias Carótidas/epidemiología , Estudios de Casos y Controles , Comorbilidad , Femenino , Humanos , Hiperhomocisteinemia/epidemiología , Hiperinsulinismo/epidemiología , Inflamación/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Medición de Riesgo , Ultrasonografía DopplerRESUMEN
Kinases have been implicated in the immunopathological mechanisms of Systemic Lupus Erythematosus (SLE). v-akt murine-thymoma viral-oncogene-homolog 1 (AKT1) and mitogen-activated-protein-kinase 1 (MAPK1) gene expressions in peripheral mononuclear cells from thirteen SLE patients with inactive or mild disease were evaluated using quantitative real-time reverse-transcription polymerase-chain-reaction and analyzed whether there was any correlation with T-helper (Th) transcription factors (TF) gene expression, cytokines, and S100A8/S100A9-(Calprotectin). Age- and gender-matched thirteen healthy controls were examined. AKT1 and MAPK1 expressions were upregulated in SLE patients and correlated with Th17-(Retinoic acid-related orphan receptor (ROR)-C), T-regulatory-(Treg)-(Transforming Growth Factor Beta (TGFB)-2), and Th2-(interleukin (IL)-5)-related genes. MAPK1 expression correlated with Th1-(IL-12A, T-box TF-(T-bet)), Th2-(GATA binding protein-(GATA)-3), and IL-10 expressions. IL-10 expression was increased and correlated with plasma Tumor Necrosis Factor (TNF)-α and Th0-(IL-2), Th1-(IL-12A, T-bet), GATA3, Treg-(Forkhead/winged-helix transcription factor- (FOXP)-3), and IL-6 expressions. FOXP3 expression, FOXP3/RORC, and FOXP3/GATA3 expression ratios were increased. Plasma IL-1ß, IL-12(p70), Interferon-(IFN)-γ, and IL-6 cytokines were augmented. Plasma IL-1ß, IL-6, IL-2, IFN-γ, TNF-α, IL-10, and IL-13 correlated with C-reactive protein, respectively. Increased Calprotectin correlated with neutrophils. Conclusion, SLE patients presented a systemic immunoinflammatory activity, augmented AKT1 and MAPK1 expressions, proinflammatory cytokines, and Calprotectin, together with increased expression of Treg-related genes, suggesting a regulatory feedback opposing the inflammatory activity.
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Interleucina-1/metabolismo , Leucocitos Mononucleares/metabolismo , Lupus Eritematoso Sistémico/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Adulto , Calgranulina A/metabolismo , Calgranulina B/metabolismo , Citocinas/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Interleucina-1/genética , Lupus Eritematoso Sistémico/sangre , Lupus Eritematoso Sistémico/genética , Masculino , Persona de Mediana Edad , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteínas Proto-Oncogénicas c-akt/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
In CD38-deficient ( Cd38-/- ) mice intraperitoneal injection of pristane induces a lupus-like disease, which is milder than that induced in WT mice, showing significant differences in the inflammatory and autoimmune processes triggered by pristane. Extracellular vesicles (EV) are present in all body fluids. Shed by cells, their molecular make-up reflects that of their cell of origin and/or tissue pathological situation. The aim of this study was to analyze the protein composition, protein abundance, and functional clustering of EV released by peritoneal exudate cells (PECs) in the pristane experimental lupus model, to identify predictive or diagnostic biomarkers that might discriminate the autoimmune process in lupus from inflammatory reactions and/or normal physiological processes. In this study, thanks to an extensive proteomic analysis and powerful bioinformatics software, distinct EV subtypes were identified in the peritoneal exudates of pristane-treated mice: 1) small EV enriched in the tetraspanin CD63 and CD9, which are likely of exosomal origin; 2) small EV enriched in CD47 and CD9, which are also enriched in plasma-membrane, membrane-associated proteins, with an ectosomal origin; 3) small EV enriched in keratins, ECM proteins, complement/coagulation proteins, fibrin clot formation proteins, and endopetidase inhibitor proteins. This enrichment may have an inflammation-mediated mesothelial-to-mesenchymal transition origin, representing a protein corona on the surface of peritoneal exudate EV; 4) HDL-enriched lipoprotein particles. Quantitative proteomic analysis allowed us to identify an anti-inflammatory, Annexin A1-enriched pro-resolving, neutrophil protein signature, which was more prominent in EV from pristane-treated Cd38-/- mice, and quantitative differences in the protein cargo of the ECM-enriched EV from Cd38-/- vs WT mice. These differences are likely to be related with the distinct inflammatory outcome shown by Cd38-/- vs WT mice in response to pristane treatment. Our results demonstrate the power of a hypothesis-free and data-driven approach to transform the heterogeneity of the peritoneal exudate EV from pristane-treated mice in valuable information about the relative proportion of different EV in a given sample and to identify potential protein markers specific for the different small EV subtypes, in particular those proteins defining EV involved in the resolution phase of chronic inflammation.
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Vesículas Extracelulares , Neutrófilos , Ratones , Animales , Proteómica , Modelos Animales de Enfermedad , Inflamación , AntiinflamatoriosRESUMEN
Global analysis of protein phosphorylation by mass spectrometry proteomic techniques has emerged in the last decades as a powerful tool in biological and biomedical research. However, there are several factors that make the global study of the phosphoproteome more challenging than measuring non-modified proteins. The low stoichiometry of the phosphorylated species and the need to retrieve residue specific information require particular attention on sample preparation, data acquisition and processing to ensure reproducibility, qualitative and quantitative robustness and ample phosphoproteome coverage in phosphoproteomic workflows. Aiming to investigate the effect of different variables in the performance of proteome wide phosphoprotein analysis protocols, ProteoRed-ISCIII and EuPA launched the Proteomics Multicentric Experiment 11 (PME11). A reference sample consisting of a yeast protein extract spiked in with different amounts of a phosphomix standard (Sigma/Merck) was distributed to 31 laboratories around the globe. Thirty-six datasets from 23 laboratories were analyzed. Our results indicate the suitability of the PME11 reference sample to benchmark and optimize phosphoproteomics strategies, weighing the influence of different factors, as well as to rank intra and inter laboratory performance.
Asunto(s)
Proteoma , Proteómica , Laboratorios , Fosfoproteínas/análisis , Fosforilación , Proteoma/análisis , Proteómica/métodos , Estándares de Referencia , Reproducibilidad de los ResultadosRESUMEN
Exosome vesicles of endocytic origin are involved in communication between tumor and immune cells. In addition, membrane rafts (MR) may support the sorting of proteins associated with exosomes. CD38 is found at the plasma membrane and in recycling endosomes, which are both redistributed toward the immunological synapse (IS) upon T cell antigen receptor (TCR) engagement. The data of this study provide evidence that CD38 is expressed on the surface of secreted exosomes derived from lymphoblastoid B cells. Exosomic CD38 is associated with the signaling molecules CD81, Hsc-70 and Lyn. Likewise, in MR, CD38 is associated with CD81, CD19, Lyn, Galphai-2, Hsc-70 and actin. Therefore, a high degree of overlap in the pattern of signaling proteins associated with CD38 in exosomes and MR exists. Exosomic and MR CD38, by virtue of these interactions, have signaling potential. Indeed, CD38 is enzymatically active in both exosomes and MR, and CD38 ligation induces Akt/PKB and Erk activation, which is accompanied by increased translocation of CD38 into MR. In conclusion, the present study indicates that CD38 localizes to MR, where it promotes cell signaling, and it is exported out of the cells through the exosome-mediated exocytic pathway, where it may act as an intercellular messenger.
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ADP-Ribosil Ciclasa 1/metabolismo , Antígenos CD/metabolismo , Linfocitos B/metabolismo , Exosomas/metabolismo , Proteínas del Choque Térmico HSC70/metabolismo , Familia-src Quinasas/metabolismo , Linfocitos B/citología , Western Blotting , Proliferación Celular , Células Cultivadas , Técnica del Anticuerpo Fluorescente , Humanos , Inmunoprecipitación , Transducción de Señal , Tetraspanina 28RESUMEN
The absence of the mouse cell surface receptor CD38 in Cd38-/- mice suggests that this receptor acts as a positive regulator of inflammatory and autoimmune responses. Here, we report that, in the context of the chronic graft-versus-host disease (cGVHD) lupus inducible model, the transfer of B6.C-H2bm12/KhEg(bm12) spleen cells into co-isogenic Cd38-/- B6 mice causes milder lupus-like autoimmunity with lower levels of anti-ssDNA autoantibodies than the transfer of bm12 spleen cells into WT B6 mice. In addition, significantly lower percentages of Tfh cells, as well as GC B cells, plasma cells, and T-bet+CD11chi B cells, were observed in Cd38-/- mice than in WT mice, while the expansion of Treg cells and Tfr cells was normal, suggesting that the ability of Cd38-/- B cells to respond to allogeneic help from bm12 CD4+ T cells is greatly diminished. The frequencies of T-bet+CD11chi B cells, which are considered the precursors of the autoantibody-secreting cells, correlate with anti-ssDNA autoantibody serum levels, IL-27, and sCD40L. Proteomics profiling of the spleens from WT cGVHD mice reflects a STAT1-driven type I IFN signature, which is absent in Cd38-/- cGVHD mice. Kidney, spleen, and liver inflammation was mild and resolved faster in Cd38-/- cGVHD mice than in WT cGVHD mice. We conclude that CD38 in B cells functions as a modulator receptor that controls autoimmune responses.
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ADP-Ribosil Ciclasa 1/deficiencia , Linfocitos B/inmunología , Linfocitos B/metabolismo , Susceptibilidad a Enfermedades , Enfermedad Injerto contra Huésped/etiología , Enfermedad Injerto contra Huésped/metabolismo , Glicoproteínas de Membrana/deficiencia , Traslado Adoptivo , Animales , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Autoinmunidad , Biomarcadores , Enfermedad Crónica , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Enfermedad Injerto contra Huésped/diagnóstico , Enfermedad Injerto contra Huésped/terapia , Inmunofenotipificación , Lupus Eritematoso Sistémico/etiología , Lupus Eritematoso Sistémico/metabolismo , Lupus Eritematoso Sistémico/patología , Recuento de Linfocitos , Ratones , Ratones Noqueados , Especificidad de Órganos , Proteoma , Proteómica/métodos , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/metabolismoRESUMEN
OBJECTIVE: compare ventilatory time between patients with the application of a disconnection protocol, managed in a coordinated way between doctor and nurse, with patients managed exclusively by the doctor. METHOD: experimental pilot study before and after. Twenty-five patients requiring invasive mechanical ventilation for 24 hours or more were included, and the protocol-guided group was compared with the protocol-free group managed according to usual practice. RESULTS: by means of the multidisciplinary protocol, the time of invasive mechanical ventilation was reduced (141.94 ± 114.50 vs 113.18 ± 55.14; overall decrease of almost 29 hours), the time spent on weaning (24 hours vs 7.40 hours) and the numbers of reintubation (13% vs 0%) in comparison with the group in which the nurse did not participate. The time to weaning was shorter in the retrospective cohort (2 days vs. 5 days), as was the hospital stay (7 days vs. 9 days). CONCLUSION: the use of a multidisciplinary protocol reduces the duration of weaning, the total time of invasive mechanical ventilation and reintubations. The more active role of the nurse is a fundamental tool to obtain better results.
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Respiración Artificial/normas , Desconexión del Ventilador/normas , Anciano , Protocolos Clínicos , Práctica Clínica Basada en la Evidencia , Femenino , Humanos , Masculino , Rol de la Enfermera , Relaciones Médico-Enfermero , Proyectos Piloto , Estudios RetrospectivosRESUMEN
The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. Complexity escalates remarkably when considering that carbohydrates can form severely branched structures with many different constituents, which often leads to the formation of multiple isomers. In this regard, ion mobility (IM) spectrometry has recently demonstrated its power for the separation of isomeric compounds. In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. Straightforward separation of isomers was achieved with the analysis of released glycans, as opposed to the glycopeptides which showed a more complex pattern. Finally, the developed methodology was applied to serum samples of mice, to investigate its robustness when analyzing real complex samples. BIOLOGICAL SIGNIFICANCE: Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer.
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Glicómica/métodos , Espectrometría de Movilidad Iónica/métodos , Isomerismo , Animales , Glicosilación , Espectrometría de Movilidad Iónica/normas , Ratones , Ácido N-Acetilneuramínico/síntesis química , Polisacáridos/análisis , Transferrina/químicaRESUMEN
In this study, we investigated the role of CD38 in a pristane-induced murine model of lupus. CD38-deficient (Cd38-/-) but not ART2-deficient (Art2-/-) mice developed less severe lupus compared to wild type (WT) mice, and their protective phenotype consisted of (i) decreased IFN-I-stimulated gene expression, (ii) decreased numbers of peritoneal CCR2hiLy6Chi inflammatory monocytes, TNF-α-producing Ly6G+ neutrophils and Ly6Clo monocytes/macrophages, (iii) decreased production of anti-single-stranded DNA and anti-nRNP autoantibodies, and (iv) ameliorated glomerulonephritis. Cd38-/- pristane-elicited peritoneal exudate cells had defective CCL2 and TNF-α secretion following TLR7 stimulation. However, Tnf-α and Cxcl12 gene expression in Cd38-/- bone marrow (BM) cells was intact, suggesting a CD38-independent TLR7/TNF-α/CXCL12 axis in the BM. Chemotactic responses of Cd38-/- Ly6Chi monocytes and Ly6G+ neutrophils were not impaired. However, Cd38-/- Ly6Chi monocytes and Ly6Clo monocytes/macrophages had defective apoptosis-mediated cell death. Importantly, mice lacking the cation channel TRPM2 (Trpm2-/-) exhibited very similar protection, with decreased numbers of PECs, and apoptotic Ly6Chi monocytes and Ly6Clo monocytes/macrophages compared to WT mice. These findings reveal a new role for CD38 in promoting aberrant inflammation and lupus-like autoimmunity via an apoptosis-driven mechanism. Furthermore, given the implications of CD38 in the activation of TRPM2, our data suggest that CD38 modulation of pristane-induced apoptosis is TRPM2-dependent.
Asunto(s)
ADP-Ribosil Ciclasa 1/metabolismo , Apoptosis , Inmunosupresores/farmacología , Lupus Eritematoso Cutáneo/inducido químicamente , Lupus Eritematoso Cutáneo/patología , Glicoproteínas de Membrana/metabolismo , Canales Catiónicos TRPM/metabolismo , Terpenos/farmacología , ADP Ribosa Transferasas/deficiencia , ADP Ribosa Transferasas/metabolismo , ADP-Ribosil Ciclasa 1/deficiencia , Animales , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Factores Inmunológicos/metabolismo , Leucocitos/inmunología , Glicoproteínas de Membrana/deficiencia , RatonesRESUMEN
In this study we have determined whether there is a relationship between CD38 expression on T cells, its distribution in different membrane microdomains, and T cell activation in SLE patients. The data show that CD38 expression is augmented in ex vivo CD3+, CD4+, CD8+, and CD25+ SLE T cells, which correlates with its increased insolubility in Brij 98 detergent, and its translocation into lipid rafts. Moreover, SLE T cells show an altered CD4:CD8 ratio, which is due to a decreased proportion of CD4+ T cells and a concomitant increase in the proportion of CD8+ T cells. These data are consistent with the increased CD38 expression and lipid raft formation, and the significant reduction in the CD4:CD8 ratio observed in mitogen-stimulated normal T cells as compared with that in ex vivo untouched normal T cells. Increased expression of CD38 in floating rafts from SLE T cells, or from activated normal T cells may modulate TCR signaling by providing or sequestering signaling molecules to the engaged TCR.
Asunto(s)
ADP-Ribosil Ciclasa 1/análisis , Lupus Eritematoso Sistémico/inmunología , Activación de Linfocitos/inmunología , Microdominios de Membrana/inmunología , Subgrupos de Linfocitos T/inmunología , ADP-Ribosil Ciclasa 1/química , Complejo CD3/análisis , Relación CD4-CD8 , Humanos , Microdominios de Membrana/química , Mitógenos/farmacología , Aceites de Plantas/química , Polietilenglicoles/química , Receptores de Antígenos de Linfocitos T/inmunología , Solubilidad , Subgrupos de Linfocitos T/efectos de los fármacosRESUMEN
CD157/Bst1 is a dual-function receptor and ß-NAD+-metabolizing ectoenzyme of the ADP-ribosyl cyclase family. Expressed in human peripheral blood neutrophils and monocytes, CD157 interacts with extracellular matrix components and regulates leukocyte diapedesis via integrin-mediated signalling in inflammation. CD157 also regulates cell migration and is a marker of adverse prognosis in epithelial ovarian cancer and pleural mesothelioma. One form of CD157 is known to date: the canonical sequence of 318 aa from a 9-exon transcript encoded by BST1 on human chromosome 4. Here we describe a second BST1 transcript, consisting of 10 exons, in human neutrophils. This transcript includes an unreported exon, exon 1b, located between exons 1 and 2 of BST1. Inclusion of exon 1b in frame yields CD157-002, a novel proteoform of 333 aa: exclusion of exon 1b by alternative splicing generates canonical CD157, the dominant proteoform in neutrophils and other tissues analysed here. In comparative functional analyses, both proteoforms were indistinguishable in cell surface localization, specific mAb binding, and behaviour in cell adhesion and migration. However, NAD glycohydrolase activity was detected in canonical CD157 alone. Comparative phylogenetics indicate that exon 1b is a genomic innovation acquired during primate evolution, pointing to the importance of alternative splicing for CD157 function.
Asunto(s)
ADP-Ribosil Ciclasa/genética , Empalme Alternativo/genética , Antígenos CD/genética , Exones/genética , Primates/genética , ADP-Ribosil Ciclasa/metabolismo , Animales , Antígenos CD/metabolismo , Secuencia de Bases , Adhesión Celular , Secuencia Conservada/genética , Evolución Molecular , Proteínas Ligadas a GPI/genética , Proteínas Ligadas a GPI/metabolismo , Células HeLa , Humanos , Neutrófilos/metabolismo , Filogenia , Isoformas de Proteínas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Superficie Celular/metabolismo , Especificidad de la Especie , Células THP-1RESUMEN
Macrophages exert potent effector functions against invading microorganisms but constitute, paradoxically, a preferential niche for many bacterial strains to replicate. Using a model of infection by Salmonella Typhimurium, we have identified a molecular mechanism regulated by the nuclear receptor LXR that limits infection of host macrophages through transcriptional activation of the multifunctional enzyme CD38. LXR agonists reduced the intracellular levels of NAD+ in a CD38-dependent manner, counteracting pathogen-induced changes in macrophage morphology and the distribution of the F-actin cytoskeleton and reducing the capability of non-opsonized Salmonella to infect macrophages. Remarkably, pharmacological treatment with an LXR agonist ameliorated clinical signs associated with Salmonella infection in vivo, and these effects were dependent on CD38 expression in bone-marrow-derived cells. Altogether, this work reveals an unappreciated role for CD38 in bacterial-host cell interaction that can be pharmacologically exploited by activation of the LXR pathway.