RESUMEN
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.
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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
Antiplatelet therapy, the gold standard of care for patients with acute coronary syndrome (ACS) undergoing percutaneous coronary intervention (PCI), is one of the therapeutic approaches most associated with the development of adverse drug reactions (ADRs). Although numerous studies have shown that pharmacological intervention based on a limited number of high-evidence variants (primarily CYP2C19*2 and *3) can reduce the incidence of major adverse cardiovascular events (MACEs), ADRs still occur at variable rates (10.1 % in our case) despite personalized therapy. This study aimed to identify novel genetic variants associated with the endpoint of MACEs 12 months after PCI by designing and analyzing a targeted gene panel. We sequenced 244 ACS-PCI-stent patients (109 with event and 135 without event) and 99 controls without structural cardiovascular disease and performed an association analysis to search for unexpected genetic variants. No single nucleotide polymorphisms reached genomic significance after correction, but three novel variants, including ABCA1 (rs2472434), KLB (rs17618244), and ZNF335 (rs3827066), may play a role in MACEs in ACS patients. These genetic variants are involved in regulating high-density lipoprotein levels and cholesterol deposition, and as they are regulatory variants, they may affect the expression of nearby lipid metabolism-related genes. Our findings suggest new targets (both at the gene and pathway levels) that may increase susceptibility to MACEs, but further research is needed to clarify the role and impact of the identified variants before these findings can be incorporated into the therapeutic decision-making process.
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Síndrome Coronario Agudo , Secuenciación de Nucleótidos de Alto Rendimiento , Intervención Coronaria Percutánea , Inhibidores de Agregación Plaquetaria , Stents , Humanos , Síndrome Coronario Agudo/genética , Síndrome Coronario Agudo/terapia , Intervención Coronaria Percutánea/efectos adversos , Masculino , Femenino , Inhibidores de Agregación Plaquetaria/uso terapéutico , Inhibidores de Agregación Plaquetaria/efectos adversos , Persona de Mediana Edad , Stents/efectos adversos , Anciano , Secuenciación de Nucleótidos de Alto Rendimiento/métodosRESUMEN
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.
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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
The wear behavior of the Mg-3wt.% Zn-0.4wt.% Ca (ZX30) alloy was tested using a pin-on-disc configuration with AZ31 alloy discs as counterparts under dry sliding conditions. The ZX30 alloy was tested in different states: as-cast, solution-treated, peak-aged, and over-aged. Wear rates and friction coefficients were measured at different loads and sliding speeds. Abrasion and oxidation were the main wear mechanisms found in all the conditions tested. Moreover, aluminum oxides were detected on the worn surfaces, which indicates the presence of an adhesive wear mechanism. The wear behavior of the studied ZX30 alloy showed a greater tendency towards oxidative wear than other Mg alloys, and the microstructure observed strongly affected the wear behavior.
RESUMEN
The microstructure and wear properties of a Mg-1wt.% Zn-1wt.% Ca (ZX11) alloy with different heat treatments have been investigated. The ZX11 alloy was tested in the as-cast state and after different heat treatment conditions: solution-treated (at 450 °C for 24 h), peak-aged (solution-treated + aged at 180 °C for 3 h), and over-aged (solution-treated + aged at 180 °C for 24 h). The microstructure of the as-cast sample showed a continuous intermetallic phase at the grain boundaries, while the heat-treated samples exhibited discrete precipitated particles within the grains. To evaluate the wear behavior, the samples were tested using a pin-on-disc configuration, where the wear rates and friction coefficients were measured at different loads and sliding speeds. An AZ31 magnesium alloy was used as the counterbody. The worn surfaces and the wear debris were studied to identify the main wear mechanisms corresponding to each test condition. The results indicated the presence of abrasion, oxidation, and adhesive wear mechanisms in all testing conditions. In the as-cast state, delamination and plastic deformation were the dominant wear mechanisms, while they were less relevant in the heat-treated conditions. The peak-aged samples exhibited the lowest wear rates, suggesting that modifying the distribution of intermetallic precipitates contributed to enhancing the wear resistance of the alloy.
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Betablockers (BBs) are prescribed for ischaemia in patients with acute coronary syndrome (ACS). In Spain, bisoprolol and carvedilol are the most prescribed BBs, but patients often had to discontinue them due to adverse effects. Single nucleotide polymorphisms (SNPs) in ADRB1, ADRB2 and CYP2D6 genes have strong evidence of pharmacogenetic association with BBs in heart failure or hypertension, but the evidence in ACS is limited. Therefore, our study focuses on investigating how these genes influence the response to BBs in ACS patients. We analysed the association between SNPs in ADRB1 Gly389Arg (rs1801253) and Ser49Gly (rs1801252), ADRB2 Gly16Arg (rs1042713) and Glu27Gln (rs1042714), and CYP2D* 6 (*2- rs1080985, *4- rs3892097, *10 - rs1065852) and the occurrence of bradycardia/hypotension events during one year of follow-up. We performed an observational study and included 285 ACS-PCI-stent patients. A first analysis including patients treated with bisoprolol and a second analysis including patients treated with other BBs were performed. We found that the presence of the G allele (Glu) of the ADRB2 gene (rs1042714; Glu27Gln) conferred a protective effect against hypotension-induced by BBs; OR (CI 95%) = 0,14 (0,03-0,60), p < 0.01. The ADRB2 (rs1042713; Gly16Arg) GG genotype could also prevent hypotensive events; OR (CI 95%) = 0.49 (0.28-0.88), p = 0015. SNPs in ADRB1 and CYP2D6 * 2, CYP2D6 * 4 weren´t associated with primary events. The effect of CYP2D6 * 10 does not seem to be relevant for the response to BBs. According to our findings, SNPs in ADRB2 (rs1042713, rs1042714) could potentially affect the response and tolerance to BBs in ACS-patients. Further studies are necessary to clarify the impact of ADRB2 polymorphisms.
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Síndrome Coronario Agudo , Hipotensión , Intervención Coronaria Percutánea , Humanos , Citocromo P-450 CYP2D6/genética , Síndrome Coronario Agudo/tratamiento farmacológico , Síndrome Coronario Agudo/genética , Bisoprolol/uso terapéutico , Antagonistas Adrenérgicos beta/uso terapéutico , Genotipo , Polimorfismo de Nucleótido Simple/genética , Receptores Adrenérgicos beta 1/genética , Receptores Adrenérgicos beta 2/genéticaRESUMEN
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
A severe form of myopia defined as pathologic/high myopia is the main cause of visual impairment and one of the most frequent causes of blindness worldwide. It is characterized by at least 6 diopters or axial length (AL) of eyeball > 26 mm and choroidal neovascularization (CNV) in 5 to 10% of cases. Ranibizumab is a humanized recombinant monoclonal antibody fragment targeted against human vascular endothelial growth factor A (VEGF-A) used in the treatment of CNV. It acts by preventing VEGF-A from interacting with its receptors (VEGFR-1 and -2) encoded by the FLT1 and KDR genes. Several studies found that the KDR and FLT1 genotypes may represent predictive determinants of efficacy in ranibizumab-treated neovascular age-related macular degeneration (nAMD) patients. We performed a retrospective study to evaluate the association of single nucleotide polymorphisms (SNPs) in VEGFR coding genes with the response rate to ranibizumab in patients with high myopia and CNV. In the association study of genotypes in FLT1 with the response to ranibizumab, we found a significant association between two FLT1 variants (rs9582036, rs7993418) with ranibizumab efficacy at the 12-month follow-up. About the KDR gene, we found that two KDR variants (rs2305948, rs2071559) are associated with best-corrected visual acuity (BCVA) improvement and KDR (rs2239702) is associated with lower rates of BCVA worsening considering a 12-month follow-up period.
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BACKGROUND: DNA methylation (DNAm) age metrics have been widely accepted as an epigenetic biomarker for biological aging and disease. The purpose of this study is to assess whether or not individuals carrying Lynch Syndrome-associated mutations are affected in their rate of biological aging, as measured by the epigenetic clock. METHODS: Genome-wide bisulfite DNA sequencing data were generated using DNA from CD4 + T-cells obtained from peripheral blood using 27 patient samples from Lynch syndrome families. Horvath's DNAm age model based on penalized linear regression was applied to estimate DNAm age from patient samples with distinct clinical and genetic characteristics to investigate cancer mutation-related aging effects. RESULTS: Both Lynch mutation carriers and controls exhibited high variability in their estimated DNAm age, but regression analysis showed steeper slope for the Lynch mutation carriers. Remarkably, six Lynch Syndrome-associated mutation carriers showed a strong correlation to the control group, and two sisters carrying Lynch Syndrome-associated mutations, with no significant difference in lifestyle and similar chronological age, were assigned very different DNAm age. CONCLUSIONS: Future studies will be required to explore, in larger patient populations, whether specific epigenetic age acceleration is predictive of time-to-cancer development, treatment response, and survival. Epigenetic clock DNAm metrics may be affected by the presence of cancer mutations in the germline, and thus show promise of potential clinical utility for stratified surveillance strategies based on the relative risk for imminent emergence of tumor lesions in otherwise healthy Lynch Syndrome-associated mutation carriers.
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Neoplasias Colorrectales Hereditarias sin Poliposis , Metilación de ADN , Aceleración , Envejecimiento/genética , Neoplasias Colorrectales Hereditarias sin Poliposis/genética , Epigénesis Genética , Humanos , MutaciónRESUMEN
Lenvatinib is an oral multityrosine kinase inhibitor (TKI) with proven effectiveness in the treatment of radioactive iodine- (RAI-) refractory and/or unresectable differentiated thyroid carcinoma (DTC). The present study reports the case of a 41-year-old male who underwent hemithyroidectomy in June 2015 due to a thyroid nodule with fine-needle aspiration follicular neoplasm cytology and no evidence of malignancy in the histopathological exam. Three years later, acute disabling clinical symptoms emerged, mainly high skeletal pain conditioned with an important performance status decrease. PET/CT scan displayed several metastatic bone lesions in this context, located in the vertebral bodies, sternum, ribs, iliac crest, right acetabulum, and both necks of the femur. Histological study and immunohistochemistry confirmed DTC metastases, as they were strongly positive for thyroglobulin and TTF-1. At this point, the patient was unfit for conventional management that would have included completion of surgery and RAI treatment as first options. Thus, it was decided to start systemic treatment with TKI, Lenvatinib. Within the first week of treatment, the patient was almost asymptomatic and his performance status moved from 3 to 0. This allowed the patient to undergo resection of the thyroid gland remnant plus RAI treatment. Unfortunately, RAI refractory illness was confirmed so Lenvatinib treatment should be continued in this case until the evidence of no further clinical benefit. Despite drug adverse events, the patient continues with treatment one year later, remaining asymptomatic and with normal functional capacity.
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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.
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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
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.
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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
We have evaluated the effect of N,N-bis (2-chloroethyl)-N-nitrosourea (BCNU), an inhibitor of glutathione reductase (GR), on the oxidative status along with the integrity of the nigrostriatal dopaminergic system of the rat. The oxidative status was studied by the quantification of carbonyl groups coupled to protein homogenates. Moreover, the specific oxidations in glial fibrillary acidic protein (GFAP) and neurofilament-200 (NF-200) were also measured. The results show that oxidative damage in proteins in the nigrostriatal system is confined to the striatum. Specific carbonyl groups coupled to native NF-200 and GFAP were also increased. These changes were accompanied by reactive astrocytosis in striatum but not in substantia nigra. In substantia nigra, decreased levels of dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) were observed following BCNU treatment. In contrast, DA levels were increased in the striatum along with an overall decrease in the ratios of DA metabolites to DA. We also studied the mRNA levels for tyrosine hydroxylase (TH) and the dopamine transporter (DAT) by in situ hybridization. TH mRNA but not DAT mRNA was significantly induced in substantia nigra following BCNU treatment, which was consistent with significant elevations in TH enzyme amount and activity and unchanged DA uptake in striatum. All these results support the DA free radical hypothesis and the key role of the striatal glutathione system in protecting the striatal system against oxidative stress.
Asunto(s)
Cuerpo Estriado/patología , Dopamina/metabolismo , Glutatión Reductasa/antagonistas & inhibidores , Glicoproteínas de Membrana , Proteínas del Tejido Nervioso , Sustancia Negra/patología , Tirosina 3-Monooxigenasa/metabolismo , Ácido 3,4-Dihidroxifenilacético/metabolismo , Animales , Western Blotting , Carmustina/farmacología , Cuerpo Estriado/metabolismo , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática , Inhibidores Enzimáticos/farmacología , Femenino , Proteína Ácida Fibrilar de la Glía/efectos de los fármacos , Proteína Ácida Fibrilar de la Glía/metabolismo , Glutatión Reductasa/efectos de los fármacos , Inmunohistoquímica , Hibridación in Situ , Proteínas de Transporte de Membrana/efectos de los fármacos , Proteínas de Transporte de Membrana/metabolismo , Proteínas de Neurofilamentos/efectos de los fármacos , Proteínas de Neurofilamentos/metabolismo , Estrés Oxidativo , Enfermedad de Parkinson/fisiopatología , Biosíntesis de Proteínas , Proteínas/metabolismo , ARN Mensajero/análisis , Ratas , Ratas Wistar , Sustancia Negra/metabolismo , Transcripción Genética , Tirosina 3-Monooxigenasa/efectos de los fármacos , Tirosina 3-Monooxigenasa/genéticaRESUMEN
Resumen ANTECEDENTES: Los linfangiomas fetales son malformaciones del sistema linfático que representan 4% de todos los tumores vasculares en los recién nacidos vivos, con una incidencia de 1.2-2.8‰. CASOS CLÍNICOS: Se comunican dos casos clínicos poco frecuentes, por su localización y extensión, de linfangiomas fetales. En ambas pacientes el diagnóstico se estableció mediante estudio ecográfico, durante el tercer trimestre, en gestaciones de bajo riesgo. La alteración no se relacionó con malformaciones estructurales adicionales, trastornos cromosómicos ni genéticos. La resonancia magnética confirmó el diagnóstico de la enfermedad. En una de las madres, el tamaño del feto determinó la vía de finalización del embarazo (parto). La cirugía fue el tratamiento de elección, con evolución satisfactoria en una paciente y la otra permanece a la espera de una nueva intervención quirúrgica, pues aún manifiesta recidivas. En la actualidad, el desarrollo psicomotor, ponderal y estructural de las pacientes es adecuado. CONCLUSIÓN: La ecografía es un estudio decisivo para establecer el diagnóstico y seguimiento de los linfangiomas fetales.
Abstract BACKGROND: Fetal lymphangiomas are malformations of the lymphatic system, representing 4% of all vascular tumors in living newborns, with an incidence of 1.2-2.8‰. CLINICAL CASE: Two uncommon clinical cases, due to their location and extension, of fetal lymphangiomas are reported. In both patients, the diagnosis was established by ultrasound study, during the third trimester, in low risk gestations. The alteration was not related to additional structural malformations, chromosomal or genetic disorders. Magnetic resonance confirmed the diagnosis of the disease. In one mothers, the size of the fetus determined the route of termination of pregnancy (delivery). Surgery was the treatment of choice, with satisfactory evolution in one patient and the other remains awaiting a new surgical intervention, because it still manifests recurrences. At present, the psychomotor, weigth and size development of the patients is adequate. CONCLUSION: Ultrasound is critical for diagnosis and monitoring of this type of malformations.
RESUMEN
CD38, a type II transmembrane glycoprotein expressed in many cells of the immune system, is involved in cell signaling, migration and differentiation. Studies in CD38 deficient mice (CD38 KO mice) indicate that this molecule controls inflammatory immune responses, although its involvement in these responses depends on the disease model analyzed. Here, we explored the role of CD38 in the control of autoimmune responses using chicken collagen type II (col II) immunized C57BL/6-CD38 KO mice as a model of collagen-induced arthritis (CIA). We demonstrate that CD38 KO mice develop an attenuated CIA that is accompanied by a limited joint induction of IL-1ß and IL-6 expression, by the lack of induction of IFNγ expression in the joints and by a reduction in the percentages of invariant NKT (iNKT) cells in the spleen. Immunized CD38 KO mice produce high levels of circulating IgG1 and low of IgG2a anti-col II antibodies in association with reduced percentages of Th1 cells in the draining lymph nodes. Altogether, our results show that CD38 participates in the pathogenesis of CIA controlling the number of iNKT cells and promoting Th1 inflammatory responses.
Asunto(s)
ADP-Ribosil Ciclasa 1/deficiencia , Artritis Experimental/inmunología , Glicoproteínas de Membrana/deficiencia , ADP-Ribosil Ciclasa 1/genética , ADP-Ribosil Ciclasa 1/inmunología , Animales , Anticuerpos Heterófilos/sangre , Artritis Experimental/genética , Artritis Experimental/patología , Pollos , Colágeno Tipo II/inmunología , Citocinas/genética , Expresión Génica , Inmunoglobulina G/sangre , Interferón gamma/biosíntesis , Interferón gamma/genética , Interleucina-1beta/genética , Interleucina-6/genética , Masculino , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Células T Asesinas Naturales/inmunología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Bazo/inmunología , Células TH1/inmunología , Células Th17/inmunologíaRESUMEN
Proteins differentially expressed in peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients versus Normal controls were identified by 2-DE and MALDI-MS. Thus, S100A9 expression was significantly increased in SLE PBMCs relative to Normal PBMCs at both mRNA and protein levels. Increased S100A9 levels in SLE PBMCs correlated positively with the abnormal presence of low-density granulocytes (LDGs) detected by flow-cytometry in the mononuclear cell fractions. Another set of proteins that were differentially expressed in SLE PBMCs formed S100A9-independent clusters, suggesting that these differences in protein expression are in fact reflecting changes in the abundance of specific cell types. In SLE PBMCs spots of the two S100A9 isoforms, S100A9-l and S100A9-s, and their phosphorylated counterparts were identified and confirmed to be phosphorylated at Thr(113) by MS/MS analyses. In addition, the phorbol ester PMA alone or in combination with ionomycin induced a stronger increase in threonine phosphorylation of S100A9 in SLE than in Normal PBMCs, while the same stimuli caused the opposite effect on phosphorylation and activation of Erk1/2, suggesting the existence of an abnormal S100A9 signaling in SLE PBMCs. Therefore, the expansion and activation of LDGs in SLE seems to underlie this prominent S100A9 signature.
Asunto(s)
Calgranulina B/biosíntesis , Leucocitos Mononucleares/metabolismo , Lupus Eritematoso Sistémico/sangre , Calgranulina B/genética , Electroforesis en Gel Bidimensional , Granulocitos/citología , Humanos , Ionomicina/farmacología , Complejo de Antígeno L1 de Leucocito/sangre , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/efectos de los fármacos , Lupus Eritematoso Sistémico/genética , Sistema de Señalización de MAP Quinasas/fisiología , Fosforilación/efectos de los fármacos , Isoformas de Proteínas/metabolismo , Proteómica , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
The aim of the present study was to explore the potential for using vermicompost from olive-mill waste as an organic amendment for enhanced bioremediation of polycyclic aromatic hydrocarbons (PAHs)-contaminated soils. The focus was to analyse the genetic potential and the naphthalene dioxygenase (NDO) expression of the bacterial communities involved in the degradation of naphthalene, as chemical model for the degradation of PAH. The structure of the metabolically active bacterial population was evidenced in the RNA-based denaturing gradient gel electrophoresis (DGGE) profiles. The relative expression of NDO was determined with real-time PCR in both the soil and the vermicompost cDNA. Naphthalene changed the structure of the metabolically active bacterial community in the vermicompost when this was artificially contaminated. When used as amendment, naphthalene-free vermicompost modified the bacterial population in the PAH-contaminated soil, evidenced in the DGGE gels after 1 month of incubation. In the amended soil, the vermicompost enhanced the NDO enzyme expression with a concomitant biodegradation of naphthalene. The effect of the vermicompost was to induce the expression of biodegradation indicator genes in the autochthonous bacterial community and/or incorporate new bacterial species capable of degrading PAH. The results indicated that vermicompost from olive-mill wastes could be considered a suitable technology to be used in PAH bioremediation.