Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 15 de 15
Filtrar
Más filtros










Base de datos
Intervalo de año de publicación
1.
Foods ; 10(3)2021 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-33808639

RESUMEN

One of the main concerns in gluten analysis is to achieve efficient extraction of gluten proteins. Conventional ethanol-based extraction solutions are inefficient and, because of this, it is necessary to use reducing agents or acids for proper solubilization. The extraction recommended by CODEX Standard 118-1979 (revised 2008) utilizes Cocktail solution (patent WO 02/092633 A1). However, it is harmful with a disgusting odor and is not compatible with some immunological techniques. Here, the versatility and extraction capacity of a new Universal Gluten Extraction Solution (UGES) (patent ES 2 392 412 A1) were evaluated using different methodological conditions, food matrices, and various immunological methods. UGES includes safer compounds for both the user and the environment, and it displayed similar extraction efficiency to that of the extraction method recommended for sandwich enzyme-linked immunosorbent assay (ELISA). The extraction time was significantly reduced from 100 to 40 min, depending on the type of the sample. Furthermore, unlike the currently used solution, UGES is compatible with competitive ELISA.

2.
World J Gastroenterol ; 25(11): 1409-1420, 2019 Mar 21.
Artículo en Inglés | MEDLINE | ID: mdl-30918433

RESUMEN

BACKGROUND: Life-long removal of gluten from the diet is currently the only way to manage celiac disease (CeD). Until now, no objective test has proven useful to objectively detect ingested gluten in clinical practice. Recently, tests that determine consumption of gluten by assessing excretion of gluten immunogenic peptides (GIP) in stool and urine have been developed. Their utility, in comparison with conventional dietary and analytical follow-up strategies, has not been fully established. AIM: To assess the performance of enzyme-linked immunosorbent assay (ELISA) and point-of-care tests (PoCTs) for GIP excretion in CeD patients on gluten-free diet (GFD). METHODS: We conducted an observational, prospective, cross-sectional study in patients following a GFD for at least two years. Using the Gastrointestinal Symptom Rating Scale questionnaire, patients were classified at enrollment as asymptomatic or symptomatic. Gluten consumption was assessed twice by 3-d dietary recall and GIP excretion (by ELISA in stool and PoCTs (commercial kits for stool and urine) in two consecutive samples. These samples and dietary reports were obtained 10 day apart one from the other. Patients were encouraged to follow their usual GFD during the study period. RESULTS: Forty-four patients were enrolled, of which 19 (43.2%) were symptomatic despite being on a GFD. Overall, 83 sets of stool and/or urine samples were collected. Eleven out of 44 patients (25.0%) had at least one positive GIP test. The occurrence of at least one positive test was 32% in asymptomatic patients compared with 15.8% in symptomatic patients. GIP was concordant with dietary reports in 65.9% of cases (Cohen´s kappa: 0.317). PoCT detected dietary indiscretions. Both ELISA and PoCT in stool were concordant (concomitantly positive or negative) in 67 out of 74 (90.5%) samples. Excretion of GIP was detected in 7 (8.4%) stool and/or urine samples from patients considered to be strictly compliant with the GFD by dietary reports. CONCLUSION: GIP detects dietary transgressions in patients on long-term GFD, irrespective of the presence of symptoms. PoCT for GIP detection constitutes a simple home-based method for self-assessment of dietary indiscretions.


Asunto(s)
Enfermedad Celíaca/dietoterapia , Dieta Sin Gluten , Glútenes/análisis , Cooperación del Paciente , Péptidos/análisis , Adulto , Anciano , Anciano de 80 o más Años , Enfermedades Asintomáticas , Enfermedad Celíaca/diagnóstico , Enfermedad Celíaca/inmunología , Enfermedad Celíaca/orina , Estudios Transversales , Autoevaluación Diagnóstica , Ensayo de Inmunoadsorción Enzimática , Heces/química , Femenino , Glútenes/química , Glútenes/inmunología , Glútenes/metabolismo , Humanos , Eliminación Intestinal , Masculino , Persona de Mediana Edad , Péptidos/química , Péptidos/inmunología , Péptidos/metabolismo , Pruebas en el Punto de Atención , Estudios Prospectivos , Encuestas y Cuestionarios
4.
Gut ; 66(2): 250-257, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-26608460

RESUMEN

OBJECTIVE: Gluten-free diet (GFD) is the only management for coeliac disease (CD). Available methods to assess GFD compliance are insufficiently sensitive to detect occasional dietary transgressions that may cause gut mucosal damage. We aimed to develop a method to determine gluten intake and monitor GFD compliance in patients with CD and to evaluate its correlation with mucosal damage. DESIGN: Urine samples of 76 healthy subjects and 58 patients with CD subjected to different gluten dietary conditions were collected. A lateral flow test (LFT) with the highly sensitive and specific G12 monoclonal antibody for the most dominant gluten immunogenic peptides (GIP) and a LFT reader were used to quantify GIP in solid-phase extracted urines. RESULTS: GIP were detectable in concentrated urines from healthy individuals previously subjected to GFD as early as 4-6 h after single gluten intake, and remained detectable for 1-2 days. The urine assay revealed infringement of the GFD in about 50% of the patients. Analysis of duodenal biopsies revealed that most of patients with CD (89%) with no villous atrophy had no detectable GIP in urine, while all patients with quantifiable GIP in urine showed incomplete intestinal mucosa recovery. CONCLUSION: GIP are detected in urine after gluten consumption, enabling a new and non-invasive method to monitor GFD compliance and transgressions. The method was sensitive, specific and simple enough to be convenient for clinical monitoring of patients with CD as well as for basic and clinical research applications including drug development. TRIAL REGISTRATION NUMBER: NCT02344758.


Asunto(s)
Enfermedad Celíaca/patología , Enfermedad Celíaca/orina , Gliadina/inmunología , Glútenes/metabolismo , Cooperación del Paciente , Péptidos/orina , Adolescente , Adulto , Anticuerpos Monoclonales , Biopsia , Estudios de Casos y Controles , Enfermedad Celíaca/dietoterapia , Niño , Preescolar , Cromatografía de Afinidad , Registros de Dieta , Dieta Sin Gluten , Duodeno/patología , Femenino , Proteínas de Unión al GTP/inmunología , Glútenes/inmunología , Humanos , Inmunoglobulina A/sangre , Masculino , Persona de Mediana Edad , Péptidos/inmunología , Proteína Glutamina Gamma Glutamiltransferasa 2 , Sensibilidad y Especificidad , Transglutaminasas/inmunología , Adulto Joven
5.
Am J Gastroenterol ; 111(10): 1456-1465, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27644734

RESUMEN

OBJECTIVES: Treatment for celiac disease (CD) is a lifelong strict gluten-free diet (GFD). Patients should be followed-up with dietary interviews and serology as CD markers to ensure adherence to the diet. However, none of these methods offer an accurate measure of dietary compliance. Our aim was to evaluate the measurement of gluten immunogenic peptides (GIP) in stools as a marker of GFD adherence in CD patients and compare it with traditional methods of GFD monitoring. METHODS: We performed a prospective, nonrandomized, multicenter study including 188 CD patients on GFD and 84 healthy controls. Subjects were given a dietary questionnaire and fecal GIP quantified by enzyme-linked immunosorbent assay (ELISA). Serological anti-tissue transglutaminase (anti-tTG) IgA and anti-deamidated gliadin peptide (anti-DGP) IgA antibodies were measured simultaneously. RESULTS: Of the 188 celiac patients, 56 (29.8%) had detectable GIP levels in stools. There was significant association between age and GIP in stools that revealed increasing dietary transgressions with advancing age (39.2% in subjects ≥13 years old) and with gender in certain age groups (60% in men ≥13 years old). No association was found between fecal GIP and dietary questionnaire or anti-tTG antibodies. However, association was detected between GIP and anti-DGP antibodies, although 46 of the 53 GIP stool-positive patients were negative for anti-DGP. CONCLUSIONS: Detection of gluten peptides in stools reveals limitations of traditional methods for monitoring GFD in celiac patients. The GIP ELISA enables direct and quantitative assessment of gluten exposure early after ingestion and could aid in the diagnosis and clinical management of nonresponsive CD and refractory CD. Trial registration number NCT02711397.


Asunto(s)
Autoanticuerpos/inmunología , Enfermedad Celíaca/dietoterapia , Registros de Dieta , Dieta Sin Gluten , Heces/química , Proteínas de Unión al GTP/inmunología , Gliadina/inmunología , Glútenes/análisis , Inmunoglobulina A/inmunología , Cooperación del Paciente , Transglutaminasas/inmunología , Adolescente , Factores de Edad , Anticuerpos/inmunología , Estudios de Casos y Controles , Enfermedad Celíaca/inmunología , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Lactante , Masculino , Estudios Prospectivos , Proteína Glutamina Gamma Glutamiltransferasa 2 , Pruebas Serológicas , Encuestas y Cuestionarios , Adulto Joven
6.
Food Chem ; 205: 36-42, 2016 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-27006211

RESUMEN

The available immunomethods for gluten quantitation could underestimate or overestimate the net immunoactivity of foods and beverages if the chosen analytical antibody is not specific to the relevant gluten immunogenic peptides (GIP). Accurate detection of the most active GIP is desirable to assess the potential celiac toxicity of food. We evaluated the capacity of the G12 monoclonal antibody for selectively depleting GIP in samples from two different gluteomes. Samples of hydrolyzed gliadin from wheat and a barley beer were used. The input (starting peptide digest of prolamins), the flow-through (unbound peptides), and the output (captured peptides) were analyzed by G12 and R5 competitive ELISA as well as by stimulation assays of T-cells from celiac patients. Most of the GIP were retained by the G12-agarose and represented the largest part of the immunogenicity of the gluten peptidome. G12 immunodepletion experiments with hydrolyzed gluten showed that this antibody reacted with those with the highest immunoactivity for celiac patients.


Asunto(s)
Antígenos/análisis , Enfermedad Celíaca/inmunología , Glútenes/inmunología , Péptidos/análisis , Péptidos/inmunología , Anticuerpos Monoclonales/inmunología , Cerveza/análisis , Ensayo de Inmunoadsorción Enzimática/métodos , Alimentos , Gliadina/química , Gliadina/metabolismo , Glútenes/metabolismo , Hordeum/química , Humanos , Hidrólisis , Prolaminas/metabolismo , Triticum/química , Triticum/inmunología
7.
PLoS One ; 8(11): e80982, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24278359

RESUMEN

We studied whether celiac disease (CD) patients produce antibodies against a novel gliadin peptide specifically generated in the duodenum of CD patients by a previously described pattern of CD-specific duodenal proteases. Fingerprinting and ion-trap mass spectrometry of CD-specific duodenal gliadin-degrading protease pattern revealed a new 8-mer gliadin-derived peptide. An ELISA against synthetic deamidated 8-mer peptides (DGP 8-mer) was used to study the presence of IgA anti-DGP 8-mer antibodies in plasma samples from 81 children (31 active CD patients (aCD), 17 CD patients on a gluten-free diet (GFD), 10 healthy controls (C) and 23 patients with other gastrointestinal pathology (GP)) and 101 adults (16 aCD, 12 GFD, 27 C and 46 GP-patients). Deamidation of the 8-mer peptide significantly increased the reactivity of the IgA antibodies from CD patients against the peptide. Significant IgA anti-DGP 8-mer antibodies levels were detected in 93.5% of aCD-, 11.8% of GFD- and 4.3% of GP-patients in children. In adults, antibodies were detected in 81.3% of aCD-patients and 8.3% of GFD-patients while were absent in 100% of C- and GP-patients. Duodenal CD-specific gliadin degrading proteases release an 8-mer gliadin peptide that once deamidated is an antigen for specific IgA antibodies in CD patients which may provide a new accurate diagnostic tool in CD.


Asunto(s)
Especificidad de Anticuerpos/inmunología , Enfermedad Celíaca/inmunología , Gliadina/inmunología , Inmunoglobulina A/inmunología , Fragmentos de Péptidos/inmunología , Adulto , Secuencia de Aminoácidos , Enfermedad Celíaca/genética , Enfermedad Celíaca/metabolismo , Enfermedad Celíaca/patología , Niño , Preescolar , Duodeno/inmunología , Duodeno/metabolismo , Ensayo de Inmunoadsorción Enzimática , Epítopos de Linfocito T/química , Epítopos de Linfocito T/inmunología , Femenino , Gliadina/química , Gliadina/metabolismo , Humanos , Inmunoglobulina A/sangre , Lactante , Masculino , Fragmentos de Péptidos/química , Péptido Hidrolasas/metabolismo , Proteolisis
8.
Bioorg Med Chem ; 20(22): 6662-8, 2012 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-23063518

RESUMEN

Bioactive natural products are a potential source of new pharmaceuticals since they offer new modes of action and more specific activities. The use of derivatization also enables the optimal structure for their biological activity to be determined. In this study several epoxycurcuphenol derivatives were synthesized. The substitution pattern on the aromatic and oxirane rings was varied along with that at the benzylic position and the length of the side chain was altered. These changes were made in order to gain a deeper understanding of the structural requirements for activity. The biological activities of these compounds were evaluated on the human leukemia cell line Jurkat using an antiproliferative assay. The activity results and structural requirements are discussed.


Asunto(s)
Sesquiterpenos/química , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Proliferación Celular/efectos de los fármacos , Óxido de Etileno/química , Humanos , Células Jurkat , Leucemia/patología , Sesquiterpenos/síntesis química , Sesquiterpenos/toxicidad , Estereoisomerismo , Relación Estructura-Actividad
9.
Clin Sci (Lond) ; 123(1): 15-27, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22214248

RESUMEN

HCV (hepatitis C virus) infection is a serious health care problem that affects more than 170 million people worldwide. Viral clearance depends on the development of a successful cellular immune response against the virus. Interestingly, such a response is altered in chronically infected patients, leading to chronic hepatitis that can result in liver fibrosis, cirrhosis and hepatocellular carcinoma. Among the mechanisms that have been described as being responsible for the immune suppression caused by the virus, Treg-cells (regulatory T-cells) are emerging as an essential component. In the present work we aim to study the effect of HCV-core protein in the development of T-cells with regulatory-like function. Using a third-generation lentiviral system to express HCV-core in CD4+ Jurkat T-cells, we describe that HCV-core-expressing Jurkat cells show an up-regulation of FOXP3 (forkhead box P3) and CTLA-4 (cytotoxic T-lymphocyte antigen-4). Moreover, we show that HCV-core-transduced Jurkat cells are able to suppress CD4+ and CD8+ T-cell responses to anti-CD3 plus anti-CD28 stimulation.


Asunto(s)
Factores de Transcripción Forkhead/metabolismo , Hepacivirus/inmunología , Antígenos de la Hepatitis C/metabolismo , Hepatitis C Crónica/inmunología , Inmunidad Celular , Linfocitos T Reguladores/metabolismo , Proteínas del Núcleo Viral/metabolismo , Western Blotting , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/virología , Antígeno CTLA-4/metabolismo , Electroforesis en Gel de Poliacrilamida , Humanos , Interleucina-2/metabolismo , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Células Jurkat , Reacción en Cadena en Tiempo Real de la Polimerasa , Linfocitos T Reguladores/virología , Transducción Genética , Regulación hacia Arriba
10.
J Autoimmun ; 38(2-3): J103-8, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22004905

RESUMEN

Pregnancy frequently has a beneficial effect on the autoimmune disease Rheumatoid Arthritis, ranging from improvement in clinical symptoms to complete remission. Despite decades of study, a mechanistic explanation remains elusive. Here, we demonstrate that an analogous pregnancy-induced remission can be observed in a mouse model of arthritis. We demonstrate that during pregnancy mice are protected from collagen-induced arthritis, but are still capable of launching normal immune responses to influenza infections. We examine the role of regulatory T (T(R)) cells in this beneficial effect. T(R) cells are essential for many aspects of immune tolerance, including the suppression of autoimmune responses. Remarkably, transfer of regulatory T cells from pregnant 'protected' mice was sufficient to confer protection to non-pregnant mice. These results suggest that regulatory T cells are responsible for the pregnancy-induced amelioration of arthritis.


Asunto(s)
Artritis Experimental/inmunología , Enfermedades Autoinmunes/inmunología , Complicaciones del Embarazo/inmunología , Linfocitos T Reguladores/inmunología , Traslado Adoptivo , Animales , Artritis Experimental/prevención & control , Enfermedades Autoinmunes/prevención & control , Femenino , Humanos , Tolerancia Inmunológica , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Embarazo
11.
Immunol Rev ; 241(1): 20-38, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21488887

RESUMEN

Antigen-presenting molecules vary between individuals of the same species, making it more difficult for pathogens to evade immune recognition and spread through the whole population. As a result of this genetic diversity, transplants between individuals are recognized as foreign and are rejected. This alloreactivity turns placental viviparity into a major immunological challenge. The maternal immune system has to balance the opposing needs of maintaining robust immune reactivity to protect both mother and fetus from invading pathogens, while at the same time tolerating highly immunogenic paternal alloantigens in order to sustain fetal integrity. Regulatory T cells are responsible for the establishment of tolerance by modulating the immune response, and uterine natural killer cells direct placentation by controlling trophoblast invasion. A variety of other cell types, including decidual stromal cells, dendritic cells, and immunomodulatory multipotent mesenchymal stromal cells, are found at the fetal-maternal interface. These cells conspire to establish a suitable environment for fetal development without compromising systemic immunity. Defects in any of these components can lead to gestational failure despite successful fertilization.


Asunto(s)
Tolerancia Inmunológica , Placentación/inmunología , Complicaciones del Embarazo/inmunología , Semen/inmunología , Linfocitos T Reguladores/inmunología , Animales , Femenino , Histocompatibilidad/inmunología , Humanos , Sistema Inmunológico , Inmunomodulación , Células Asesinas Naturales/inmunología , Intercambio Materno-Fetal , Embarazo/inmunología , Complicaciones del Embarazo/prevención & control
12.
J Immunol ; 186(10): 5569-79, 2011 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-21464089

RESUMEN

IL-10-producing B cells, also known as regulatory B cells (Bregs), play a key role in controlling autoimmunity. In this study, we report that chimeric mice specifically lacking IL-10-producing B cells (IL-10(-/-)B cell) developed an exacerbated arthritis compared with chimeric wild-type (WT) B cell mice. A significant decrease in the absolute numbers of Foxp3 regulatory T cells (Tregs), in their expression level of Foxp3, and a marked increase in inflammatory Th1 and Th17 cells were detected in IL-10(-/-) B cell mice compared with WT B cell mice. Reconstitution of arthritic B cell deficient (µMT) mice with different B cell subsets revealed that the ability to modulate Treg frequencies in vivo is exclusively restricted to transitional 2 marginal zone precursor Bregs. Moreover, transfer of WT transitional 2 marginal zone precursor Bregs to arthritic IL-10(-/-) mice increased Foxp3(+) Tregs and reduced Th1 and Th17 cell frequencies to levels measured in arthritic WT mice and inhibited inflammation. In vitro, IL-10(+/+) B cells established longer contact times with arthritogenic CD4(+)CD25(-) T cells compared with IL-10(-/-) B cells in response to Ag stimulation, and using the same culture conditions, we observed upregulation of Foxp3 on CD4(+) T cells. Thus, IL-10-producing B cells restrain inflammation by promoting differentiation of immunoregulatory over proinflammatory T cells.


Asunto(s)
Artritis/inmunología , Subgrupos de Linfocitos B/inmunología , Interleucina-10/inmunología , Linfocitos T Reguladores/inmunología , Células TH1/inmunología , Células Th17/inmunología , Traslado Adoptivo , Animales , Artritis/genética , Autoinmunidad/genética , Antígenos CD4/genética , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Factores de Transcripción Forkhead/genética , Inflamación/inmunología , Interferón gamma/metabolismo , Interleucina-10/deficiencia , Interleucina-10/metabolismo , Interleucina-17/inmunología , Interleucina-17/metabolismo , Subunidad alfa del Receptor de Interleucina-2/genética , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Células Precursoras de Linfocitos B/inmunología , Linfocitos T Reguladores/citología , Linfocitos T Reguladores/metabolismo
13.
J Leukoc Biol ; 89(1): 63-73, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-20940326

RESUMEN

The adaptor protein LAT has a prominent role in the transduction of intracellular signals elicited by the TCR/CD3 complex. Upon TCR engagement, LAT becomes tyrosine-phosphorylated and thereby, recruits to the membrane several proteins implicated in the activation of downstream signaling pathways. However, little is known about the role of other conserved motifs present in the LAT sequence. Here, we report that the adaptor LAT contains several conserved serine-based motifs, which are essential for proper signal transduction through the TCR. Mutation of these serine motifs in the human T cell line Jurkat prevents proper calcium influx, MAPK activation, and IL-2 production in response to TCR/CD3 stimulation. Moreover, this mutant form of LAT has a reduced ability to bind to PLC-γ1 and SLP-76, although phosphorylation of tyrosine residues 132, 171, and 191 is not decreased, raising a possible role for the serine-based motifs of LAT for the binding of important partners. The functional role of LAT serine-based motifs in signal transduction could be mediated by an effect on tyrosine phosphorylation, as their mutation significantly diminishes the phosphorylation of tyrosine residue 226. In addition, these serine motifs seem to have a regulatory role, given that upon their mutation, ZAP-70 shows enhanced phosphorylation. Therefore, the LAT serine-based motifs likely regulate signaling pathways that are essential for T cell physiology.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/química , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas de la Membrana/química , Proteínas de la Membrana/metabolismo , Receptores de Antígenos de Linfocitos T/metabolismo , Serina/metabolismo , Transducción de Señal/inmunología , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Calcio/metabolismo , Membrana Celular/metabolismo , Secuencia Conservada/genética , Activación Enzimática , Células HEK293 , Humanos , Interleucina-2/biosíntesis , Células Jurkat , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Datos de Secuencia Molecular , Proteínas Mutantes/metabolismo , Mutación/genética , Fosfolipasa C gamma/metabolismo , Fosfoproteínas/metabolismo , Fosfotirosina/metabolismo , Unión Proteica , Relación Estructura-Actividad , Transfección , Proteína Tirosina Quinasa ZAP-70/metabolismo
14.
Transplantation ; 88(3 Suppl): S24-30, 2009 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-19667958

RESUMEN

BACKGROUND: The search for new immunosuppressive drugs is a high priority. Sesquiterpenes constitute a family of compounds with a great variety of biological activities due to their reactive moieties. METHODS: Human tumor cell lines and murine primary cells and human peripheral blood mononuclear cells or primary CD4+ cells from healthy individuals were stimulated in the presence of sesquiterpenes. Cell division was analyzed by 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester, cell cycle progression by Hoecht, and cell death by Anexin-V and propidium iodine staining. Cytokine secretion was analyzed by means of a bioplex assay. RESULTS: Two sesquiterpene derivatives of the 18 previously shown to inhibit vegetal cell growth are shown to block cell division and cell cycle progression in human and murine cell lines and primary cells. Cytokine secretion is also impaired on stimulation in the presence of sesquiterpenes. CONCLUSIONS: Here, we show that sesquiterpenes heliannuols constitute a novel family of molecules with potential use as immunosuppressants. Moreover, we show that an assay based on the allelopathic effect of plant leads can be used as a cost-effective screening previous to studies in mammalian cells.


Asunto(s)
Inmunosupresores/farmacología , Sesquiterpenos/farmacología , Animales , Antígenos CD4/inmunología , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Ciclo Celular/efectos de los fármacos , Ciclo Celular/inmunología , Muerte Celular/efectos de los fármacos , División Celular/efectos de los fármacos , División Celular/inmunología , Línea Celular Tumoral/inmunología , Citocinas/inmunología , Citocinas/metabolismo , Helianthus , Humanos , Células Jurkat/efectos de los fármacos , Células Jurkat/inmunología , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Linfocitos/efectos de los fármacos , Linfocitos/inmunología , Ratones , Extractos Vegetales/uso terapéutico
15.
Mol Immunol ; 45(7): 1863-71, 2008 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-18078995

RESUMEN

The nuclear factor of activated T cells (NFAT) family of transcription factors is pivotal for T lymphocyte functionality. All relevant NFAT activation events upon T cells stimulation such as nuclear translocation, DNA binding, and transcriptional activity have been shown to be dictated by its phosphorylation state. Here, we provide evidence for a novel post-translational modification that regulates NFAT. Indeed, NFATc1 and NFATc2 are poly(ADP-ribosyl)ated by poly-ADP-ribose polymerase-1 (PARP-1). Moreover, we have also found a physical interaction between PARP-1 and both NFATc1 and NFATc2. Interestingly, PARP is activated during T cell stimulation in the absence of DNA damage, leading to ADP-ribose polymers formation and transfer to nuclear acceptor proteins. Our data suggest that poly(ADP-ribosyl)ation modulates the activation of NFAT in T cells, as PARP inhibition causes an increase in NFAT-dependent transactivation and a delay in NFAT nuclear export. Poly(ADP-ribosyl)ation will expedited NFAT export from the nucleus directly or by priming/facilitating NFAT phosphorylation. Altogether, these data point to PARP-1 and poly(ADP-ribosyl)ation as a novel regulatory mechanism of NFAT at nuclear level, suggesting a potential use of PARP as a new therapeutic target in the modulation of NFAT.


Asunto(s)
Factores de Transcripción NFATC/metabolismo , Poli(ADP-Ribosa) Polimerasas/metabolismo , Linfocitos T/enzimología , Transporte Activo de Núcleo Celular/efectos de los fármacos , Adulto , Animales , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Inducción Enzimática/efectos de los fármacos , Citometría de Flujo , Células HeLa , Histonas/metabolismo , Humanos , Ionomicina/farmacología , Células Jurkat , Activación de Linfocitos/efectos de los fármacos , Ratones , Fosforilación/efectos de los fármacos , Poli Adenosina Difosfato Ribosa/metabolismo , Inhibidores de Poli(ADP-Ribosa) Polimerasas , Unión Proteica/efectos de los fármacos , Especificidad por Sustrato/efectos de los fármacos , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacología , Activación Transcripcional/efectos de los fármacos
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA