RESUMEN
There is a growing awareness that transient, sublethal embryonic exposure to crude oils cause subtle but important forms of delayed toxicity in fish. While the precise mechanisms for this loss of individual fitness are not well understood, they involve the disruption of early cardiogenesis and a subsequent pathological remodeling of the heart much later in juveniles. This developmental cardiotoxicity is attributable, in turn, to the inhibitory actions of crude oil-derived mixtures of polycyclic aromatic compounds (PACs) on specific ion channels and other proteins that collectively drive the rhythmic contractions of heart muscle cells via excitation-contraction coupling. Here we exposed Pacific herring (Clupea pallasi) embryos to oiled gravel effluent yielding ΣPAC concentrations as low as ~ 1 µg/L (64 ng/g in tissues). Upon hatching in clean seawater, and following the depuration of tissue PACs (as evidenced by basal levels of cyp1a gene expression), the ventricles of larval herring hearts showed a concentration-dependent reduction in posterior growth (ballooning). This was followed weeks later in feeding larvae by abnormal trabeculation, or formation of the finger-like projections of interior spongy myocardium, and months later with hypertrophy (overgrowth) of the spongy myocardium in early juveniles. Given that heart muscle cell differentiation and migration are driven by Ca2+-dependent intracellular signaling, the observed disruption of ventricular morphogenesis was likely a secondary (downstream) consequence of reduced calcium cycling and contractility in embryonic cardiomyocytes. We propose defective trabeculation as a promising phenotypic anchor for novel morphometric indicators of latent cardiac injury in oil-exposed herring, including an abnormal persistence of cardiac jelly in the ventricle wall and cardiomyocyte hyperproliferation. At a corresponding molecular level, quantitative expression assays in the present study also support biomarker roles for genes known to be involved in muscle contractility (atp2a2, myl7, myh7), cardiomyocyte precursor fate (nkx2.5) and ventricular trabeculation (nrg2, and hbegfa). Overall, our findings reinforce both proximal and indirect roles for dysregulated intracellular calcium cycling in the canonical fish early life stage crude oil toxicity syndrome. More work on Ca2+-mediated cellular dynamics and transcription in developing cardiomyocytes is needed. Nevertheless, the highly specific actions of ΣPAC mixtures on the heart at low, parts-per-billion tissue concentrations directly contravene classical assumptions of baseline (i.e., non-specific) crude oil toxicity.
Asunto(s)
Petróleo/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Cardiotoxicidad/patología , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/patología , Peces/embriología , Peces/fisiología , Corazón , Larva , Miocardio/química , Contaminación por Petróleo , Hidrocarburos Policíclicos Aromáticos/toxicidad , Agua de MarRESUMEN
As Arctic ice recedes, future oil spills pose increasing risk to keystone species and the ecosystems they support. We show that Polar cod (Boreogadus saida), an energy-rich forage fish for marine mammals, seabirds, and other fish, are highly sensitive to developmental impacts of crude oil. Transient oil exposures ≥300 µg/L during mid-organogenesis disrupted the normal patterning of the jaw as well as the formation and function of the heart, in a manner expected to be lethal to post-hatch larvae. More importantly, we found that exposure to lower levels of oil caused a dysregulation of lipid metabolism and growth that persisted in morphologically normal juveniles. As lipid content is critical for overwinter survival and recruitment, we anticipate Polar cod losses following Arctic oil spills as a consequence of both near-term and delayed mortality. These losses will likely influence energy flow within Arctic food webs in ways that are as-yet poorly understood.
RESUMEN
Passive sampling devices were used to measure air vapor and water dissolved phase concentrations of 33 polycyclic aromatic hydrocarbons (PAHs) and 22 oxygenated PAHs (OPAHs) at four Gulf of Mexico coastal sites prior to, during and after shoreline oiling from the Deepwater Horizon oil spill (DWH). Measurements were taken at each site over a 13 month period, and flux across the water-air boundary was determined. This is the first report of vapor phase and diffusive flux of both PAHs and OPAHs during the DWH. Vapor phase sum PAH and OPAH concentrations ranged between 6.6 and 210 ng/m(3) and 0.02 and 34 ng/m(3) respectively. PAH and OPAH concentrations in air exhibited different spatial and temporal trends than in water, and air-water flux of 13 individual PAHs was shown to be at least partially influenced by the DWH incident. The largest PAH volatilizations occurred at the sites in Alabama and Mississippi at nominal rates of 56â¯000 and 42â¯000 ng/m(2) day(-1) in the summer. Naphthalene was the PAH with the highest observed volatilization rate of 52â¯000 ng/m(2) day(-1) in June 2010. This work represents additional evidence of the DWH incident contributing to air contamination, and provides one of the first quantitative air-water chemical flux determinations with passive sampling technology.
Asunto(s)
Monitoreo del Ambiente , Contaminantes Químicos del Agua , Contaminación por Petróleo , Hidrocarburos Policíclicos Aromáticos , AguaRESUMEN
Passive sampling devices were used to measure air vapor and water dissolved phase concentrations of 33 polycyclic aromatic hydrocarbons (PAHs) and 22 oxygenated PAHs (OPAHs) at four Gulf of Mexico coastal sites prior to, during, and after shoreline oiling from the Deepwater Horizon oil spill (DWH). Measurements were taken at each site over a 13 month period, and flux across the water-air boundary was determined. This is the first report of vapor phase and flux of both PAHs and OPAHs during the DWH. Vapor phase sum PAH and OPAH concentrations ranged between 1 and 24 ng/m(3) and 0.3 and 27 ng/m(3), respectively. PAH and OPAH concentrations in air exhibited different spatial and temporal trends than in water, and air-water flux of 13 individual PAHs were strongly associated with the DWH incident. The largest PAH volatilizations occurred at the sites in Alabama and Mississippi in the summer, each nominally 10,000 ng/m(2)/day. Acenaphthene was the PAH with the highest observed volatilization rate of 6800 ng/m(2)/day in September 2010. This work represents additional evidence of the DWH incident contributing to air contamination, and provides one of the first quantitative air-water chemical flux determinations with passive sampling technology.
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Contaminantes Atmosféricos/análisis , Contaminación por Petróleo/análisis , Hidrocarburos Policíclicos Aromáticos/análisis , Contaminantes Químicos del Agua/análisis , Contaminantes Atmosféricos/química , Alabama , Monitoreo del Ambiente , Mississippi , Oxígeno/química , Hidrocarburos Policíclicos Aromáticos/química , Estaciones del Año , Sudeste de Estados Unidos , Volatilización , Agua , Contaminantes Químicos del Agua/químicaRESUMEN
Sequestering semi-polar compounds can be difficult with low-density polyethylene (LDPE), but those pollutants may be more efficiently absorbed using silicone. In this work, optimized methods for cleaning, infusing reference standards, and polymer extraction are reported along with field comparisons of several silicone materials for polycyclic aromatic hydrocarbons (PAHs) and pesticides. In a final field demonstration, the most optimal silicone material is coupled with LDPE in a large-scale study to examine PAHs in addition to oxygenated-PAHs (OPAHs) at a Superfund site. OPAHs exemplify a sensitive range of chemical properties to compare polymers (log Kow 0.2-5.3), and transformation products of commonly studied parent PAHs. On average, while polymer concentrations differed nearly 7-fold, water-calculated values were more similar (about 3.5-fold or less) for both PAHs (17) and OPAHs (7). Individual water concentrations of OPAHs differed dramatically between silicone and LDPE, highlighting the advantages of choosing appropriate polymers and optimized methods for pollutant monitoring.
Asunto(s)
Monitoreo del Ambiente/instrumentación , Hidrocarburos Policíclicos Aromáticos/análisis , Polietileno/química , Siliconas/química , Contaminantes Químicos del Agua/análisisRESUMEN
The authors investigated coupling passive sampling technologies with ultraviolet irradiation experiments to study polycyclic aromatic hydrocarbon (PAH) and oxygenated PAH transformation processes in real-world bioavailable mixtures. Passive sampling device (PSD) extracts were obtained from coastal waters impacted by the Deepwater Horizon oil spill and Superfund sites in Portland, Oregon, USA. Oxygenated PAHs were found in the contaminated waters with our PSDs. All mixtures were subsequently exposed to a mild dose of ultraviolet B (UVB). A reduction in PAH levels and simultaneous formation of several oxygenated PAHs were measured. Site-specific differences were observed with UVB-exposed PSD mixtures.
Asunto(s)
Monitoreo del Ambiente/métodos , Hidrocarburos Policíclicos Aromáticos/análisis , Rayos Ultravioleta , Contaminantes Químicos del Agua/análisis , Monitoreo del Ambiente/instrumentación , Golfo de México , Oregon , Oxígeno/química , Contaminación por Petróleo , Fotólisis , Hidrocarburos Policíclicos Aromáticos/química , Hidrocarburos Policíclicos Aromáticos/efectos de la radiación , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/efectos de la radiaciónRESUMEN
Biological Response Indicator Devices Gauging Environmental Stressors (BRIDGES) is a bioanalytical tool that combines passive sampling with the embryonic zebrafish developmental toxicity bioassay to provide a quantitative measure of the toxicity of bioavailable complex mixtures. Passive sampling devices (PSDs), which sequester and concentrate bioavailable organic contaminants from the environment, were deployed in the Willamette and Columbia Rivers within and outside of the Portland Harbor Superfund site in Portland, OR, USA. Six sampling events were conducted in the summer and fall of 2009 and 2010. Passive sampling device extracts were analyzed for polycyclic aromatic hydrocarbon (PAH) compounds and screened for 1,201 chemicals of concern using deconvolution-reporting software. The developmental toxicity of the extracts was analyzed using the embryonic zebrafish bioassay. The BRIDGES tool provided site-specific, temporally resolved information about environmental contaminant mixtures and their toxicity. Multivariate modeling approaches were applied to paired chemical and toxic effects data sets to help unravel chemistry-toxicity associations. Modeling elucidated spatial and temporal trends in PAH concentrations and the toxicity of the samples and identified a subset of PAH analytes that were the most highly correlated with observed toxicity. Although the present study highlights the complexity of discerning specific bioactive compounds in complex mixtures, it demonstrates methods for associating toxic effects with chemical characteristics of environmental samples.
Asunto(s)
Monitoreo del Ambiente/métodos , Contaminantes Químicos del Agua/toxicidad , Bioensayo , Monitoreo del Ambiente/instrumentación , Hidrocarburos Policíclicos Aromáticos/análisis , Hidrocarburos Policíclicos Aromáticos/toxicidad , Ríos/química , Contaminantes Químicos del Agua/análisisRESUMEN
The use of passive sampling devices (PSDs) for monitoring hydrophobic organic contaminants in aquatic environments can entail logistical constraints that often limit a comprehensive statistical sampling plan, thus resulting in a restricted number of samples. The present study demonstrates an approach for using the results of a pilot study designed to estimate sampling variability, which in turn can be used as variance estimates for confidence intervals for future n = 1 PSD samples of the same aquatic system. Sets of three to five PSDs were deployed in the Portland Harbor Superfund site for three sampling periods over the course of two years. The PSD filters were extracted and, as a composite sample, analyzed for 33 polycyclic aromatic hydrocarbon compounds. The between-sample and within-sample variances were calculated to characterize sources of variability in the environment and sampling methodology. A method for calculating a statistically reliable and defensible confidence interval for the mean of a single aquatic passive sampler observation (i.e., n = 1) using an estimate of sample variance derived from a pilot study is presented. Coverage probabilities are explored over a range of variance values using a Monte Carlo simulation.
Asunto(s)
Monitoreo del Ambiente/métodos , Hidrocarburos Policíclicos Aromáticos/análisis , Contaminantes Químicos del Agua/análisis , Contaminación Química del Agua/estadística & datos numéricos , Intervalos de Confianza , Monitoreo del Ambiente/instrumentación , Interacciones Hidrofóbicas e Hidrofílicas , Proyectos Piloto , Hidrocarburos Policíclicos Aromáticos/química , Contaminantes Químicos del Agua/químicaAsunto(s)
Enfermedades Metabólicas/complicaciones , Obesidad/complicaciones , Adiposidad/genética , Adolescente , Biomarcadores/metabolismo , Congresos como Asunto , Femenino , Humanos , Masculino , Enfermedades Metabólicas/tratamiento farmacológico , Enfermedades Metabólicas/epidemiología , Enfermedades Metabólicas/genética , Obesidad/tratamiento farmacológico , Obesidad/epidemiología , Obesidad/genética , Especificidad de la Especie , Adulto JovenRESUMEN
An estimated 4.1 million barrels of oil and 2.1 million gallons of dispersants were released into the Gulf of Mexico during the Deepwater Horizon oil spill. There is a continued need for information about the impacts and long-term effects of the disaster on the Gulf of Mexico. The objectives of this study were to assess bioavailable polycyclic aromatic hydrocarbons (PAHs) in the coastal waters of four Gulf Coast states that were impacted by the spill. For over a year, beginning in May 2010, passive sampling devices were used to monitor the bioavailable concentration of PAHs. Prior to shoreline oiling, baseline data were obtained at all the study sites, allowing for direct before and after comparisons of PAH contamination. Significant increases in bioavailable PAHs were seen following the oil spill, however, preoiling levels were observed at all sites by March 2011. A return to elevated PAH concentrations, accompanied by a chemical fingerprint similar to that observed while the site was being impacted by the spill, was observed in Alabama in summer 2011. Chemical forensic modeling demonstrated that elevated PAH concentrations are associated with distinctive chemical profiles.
Asunto(s)
Contaminación por Petróleo , Hidrocarburos Policíclicos Aromáticos/análisis , Contaminantes Químicos del Agua/análisis , Monitoreo del Ambiente , Golfo de MéxicoRESUMEN
Passive sampling devices (PSDs) sequester the freely dissolved fraction of lipophilic contaminants, mimicking passive chemical uptake and accumulation by biomembranes and lipid tissues. Public Health Assessments that inform the public about health risks from exposure to contaminants through consumption of resident fish are generally based on tissue data, which can be difficult to obtain and requires destructive sampling. The purpose of this study is to apply PSD data in a Public Health Assessment to demonstrate that PSDs can be used as a biological surrogate to evaluate potential human health risks and elucidate spatio-temporal variations in risk. PSDs were used to measure polycyclic aromatic hydrocarbons (PAHs) in the Willamette River; upriver, downriver and within the Portland Harbor Superfund megasite for 3 years during wet and dry seasons. Based on an existing Public Health Assessment for this area, concentrations of PAHs in PSDs were substituted for fish tissue concentrations. PSD measured PAH concentrations captured the magnitude, range and variability of PAH concentrations reported for fish/shellfish from Portland Harbor. Using PSD results in place of fish data revealed an unacceptable risk level for cancer in all seasons but no unacceptable risk for non-cancer endpoints. Estimated cancer risk varied by several orders of magnitude based on season and location. Sites near coal tar contamination demonstrated the highest risk, particularly during the dry season and remediation activities. Incorporating PSD data into Public Health Assessments provides specific spatial and temporal contaminant exposure information that can assist public health professionals in evaluating human health risks.
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Monitoreo del Ambiente/instrumentación , Modelos Estadísticos , Hidrocarburos Policíclicos Aromáticos/análisis , Hidrocarburos Policíclicos Aromáticos/toxicidad , Salud Pública/estadística & datos numéricos , Medición de Riesgo/métodos , Ríos/química , Animales , Peces , Humanos , Neoplasias/inducido químicamente , Oregon , Factores de Tiempo , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidadRESUMEN
BACKGROUND: Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) is a primary immunodeficiency with autoimmunity caused by mutations in forkhead box P3 (FOXP3), which encodes a transcription factor involved in regulatory T (Treg) cell function. The mechanistic basis for how different mutations in FOXP3 cause distinct manifestations of IPEX remains unclear. OBJECTIVE: To determine whether 3 different point mutants of FOXP3 that cause severe or mild IPEX differ in their ability to reprogram conventional T cells into Treg cells. METHODS: Human CD4(+) T cells were transduced with wild-type or point mutant forms of FOXP3, and changes in cell surface marker expression, cytokine production, proliferation and suppressive capacity were assessed. Ex vivo T(H)17 cells were also transduced with different forms of FOXP3 to monitor changes in IL-17 production. RESULTS: The forkhead mutant F373A failed to upregulate CD25 and CCR4, did not suppress cytokine production, and induced suppressive activity less effectively than wild-type FOXP3. In contrast, although the forkhead mutant R347H was also defective in upregulation of CD25, it suppressed the production of cytokines, conferred suppressive capacity on CD4(+) T cells, and suppressed IL-17 production. F324L, a mutant outside the forkhead domain associated with mild IPEX, was equivalent to wild-type FOXP3 in all aspects tested. CONCLUSION: Mutations in FOXP3 that cause IPEX do not uniformly abrogate the ability of FOXP3 to regulate transcription and drive the development of Treg cells. These data support the notion that factors in addition to functional changes in Treg cells, such as alterations in conventional T cells, are involved in the pathogenesis of IPEX.
Asunto(s)
Factores de Transcripción Forkhead/metabolismo , Síndromes de Inmunodeficiencia/genética , Proteínas Mutantes/metabolismo , Subgrupos de Linfocitos T/metabolismo , Linfocitos T Reguladores/metabolismo , Adolescente , Antígenos CD4/biosíntesis , Proliferación Celular , Transdiferenciación Celular/genética , Células Cultivadas , Niño , Citocinas/genética , Citocinas/metabolismo , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/inmunología , Humanos , Síndromes de Inmunodeficiencia/inmunología , Síndromes de Inmunodeficiencia/patología , Síndromes de Inmunodeficiencia/fisiopatología , Terapia de Inmunosupresión , Proteínas Mutantes/genética , Proteínas Mutantes/inmunología , Mutación Puntual/genética , Poliendocrinopatías Autoinmunes , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/patología , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/patología , Transgenes/genética , Adulto JovenRESUMEN
Engineered skin substitutes (ESSs) comprising both keratinocytes and fibroblasts can afford many advantages over the use of autologous keratinocyte grafts for the treatment of full-thickness and partial-thickness burns. In this study, we investigated the efficacy of a novel ESS containing both genetically altered fibroblasts that express the immunosuppressive factor indoleamine 2,3-dioxygenase (IDO) and primary keratinocytes from a nonautologous source to confer immune protection of xenogeneic cells cultured in a bilayer ESS. The results show that engraftment of IDO expressing skin substitutes on the back of rats significantly improves healing progression over 7 days compared with both nontreated and non-IDO-expressing skin substitutes (p<0.001). Immuno-staining of CD3 and CD31 suggests that IDO-expressing skin substitutes significantly suppress T cell infiltration (p<0.001) and improve neovascularization by four-fold (12.6±1.2 vs. 3.0±1.0 vessel-like structure/high power field), respectively. In conclusion, we found that IDO expression can improve the efficacy of nonautologous ESS for the purpose of wound healing by mitigating T-cell infiltration as well as promoting vascularization of the graft.
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Fibroblastos/metabolismo , Rechazo de Injerto/inmunología , Inmunosupresores/metabolismo , Indolamina-Pirrol 2,3,-Dioxigenasa/metabolismo , Piel Artificial , Linfocitos T/metabolismo , Adenoviridae/genética , Animales , Proliferación Celular , Células Cultivadas , Vectores Genéticos , Inmunosupresores/farmacología , Indolamina-Pirrol 2,3,-Dioxigenasa/farmacología , Neovascularización Fisiológica , Ratas , Ratas Sprague-Dawley , Ingeniería de Tejidos , Transfección , Cicatrización de HeridasRESUMEN
In the realm of multi-dimensional confocal microscopy, colocalization analysis of fluorescent emission signals has proven to be an invaluable tool for detecting molecular interactions between biological macromolecules at the subcellular level. We show here that image processing operations such as the deconvolution and chromatic corrections play a crucial role in the accurate determination of colocalization between biological macromolecules particularly when the fluorescent signals are faint, and when the fluorescent signals are in the blue and red emission regions. The cellular system presented here describes quantification of an activated forkhead box P3 (FOXP3) transcription factor in three-dimensional (3D) cellular space. 293T cells transfected with a conditionally active form of FOXP3 were stained for anti-FOXP3 conjugated to a fluorescent red dye (Phycoerythrin), and counterstained for DNA (nucleus) with fluorescent blue dye (Hoechst). Due to the broad emission spectra of these dyes, the fluorescent signals were collected only from peak regions and were acquired sequentially. Since the PE signal was weak, a confocal pinhole size of two Airy size was used to collect the 3D image data sets. The raw images supplemented with the spectral data show the preferential association of activated FOXP3 molecules with the nucleus. However, the PE signals were found to be highly diffusive and colocalization quantification from these raw images was not possible. In order to deconvolve the 3D raw image data set, point spread functions (PSFs) of these emissions were measured. From the measured PSF, we found that chromatic shifts between the blue and red colors were quite considerable. Followed by the applications of both the axial and lateral chromatic corrections, colocalization analysis performed on the deconvolved-chromatic corrected-3D image data set showed that 98% of DNA molecules were associated with FOXP3 molecules, whereas only 66% of FOXP3 molecules were colocalized with DNA molecules. In conclusion, our studies clearly demonstrate the importance of PSF measurements, chromatic aberration corrections followed by deconvolution in the accurate determination of transcription factors in the 3D cellular space. The reported imaging and processing methods can be a practical guide for quantitative fluorescence imaging of similar cellular systems and can provide a basis for further development.
Asunto(s)
Citosol/química , Procesamiento de Imagen Asistido por Computador/métodos , Imagenología Tridimensional/métodos , Microscopía Confocal/métodos , Microscopía Fluorescente/métodos , Factores de Transcripción/análisis , Línea Celular , HumanosRESUMEN
The Biological Response Indicator Devices Gauging Environmental Stressors (BRIDGES) bio-analytical tool was developed in response to the need for a quantitative technology for assessing the toxicity of environmentally relevant contaminant mixtures. This tool combines passive samplers with the embryonic zebrafish model. When applied in an urban river it effectively linked site specific, bioavailable contaminant mixtures to multiple biological responses. Embryonic zebrafish exposed to extracts from lipid-free passive samplers that were deployed at five locations, within and outside of the Portland Harbor Superfund Megasite, displayed different responses. Six of the eighteen biological responses observed in 941 exposed zebrafish were significantly different between sites. This demonstrates the sensitivity of the bio-analytical tool for detecting spatially distinct toxicity in aquatic systems; bridging environmental exposure to biological response.
Asunto(s)
Contaminantes Químicos del Agua/toxicidad , Pez Cebra/embriología , Animales , Embrión no Mamífero/efectos de los fármacos , Desarrollo Embrionario , Monitoreo del Ambiente , Lípidos/química , Ríos/químicaRESUMEN
FOXP3 is required for the development of Treg and its expression is often used as a surrogate marker of functional suppression. However, it is now known that activated human T effector cells can also express FOXP3 without acquiring regulatory activity. To more closely examine the requirements for FOXP3 to reprogram human T cells into Treg, we developed a conditionally active form of FOXP3 and show here that full acquisition of Treg phenotype and function is strictly dependent on the amount of active FOXP3 a T cell expresses. In addition, the phenotypic and functional alterations induced by FOXP3 are only fully manifested following prolonged induction of protein activity. Induction of FOXP3 activity does not upregulate EBI3 or p35 mRNA, providing evidence that secretion of IL-35 does not substantially contribute to the suppressive mechanism of human Treg. These data represent the first formal evidence that FOXP3 acts as a quantitative regulator rather than a simple molecular switch for Treg.
Asunto(s)
Diferenciación Celular/inmunología , Factores de Transcripción Forkhead/inmunología , Linfocitos T/citología , Linfocitos T/inmunología , Células Cultivadas , Citocinas/inmunología , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Humanos , Linfocitos T/metabolismo , Factores de TiempoRESUMEN
SUMMARY: T-regulatory cells (Tregs) have a fundamental role in the establishment and maintenance of peripheral tolerance. There is now compelling evidence that deficits in the numbers and/or function of different types of Tregs can lead to autoimmunity, allergy, and graft rejection, whereas an over-abundance of Tregs can inhibit anti-tumor and anti-pathogen immunity. Experimental models in mice have demonstrated that manipulating the numbers and/or function of Tregs can decrease pathology in a wide range of contexts, including transplantation, autoimmunity, and cancer, and it is widely assumed that similar approaches will be possible in humans. Research into how Tregs can be manipulated therapeutically in humans is most advanced for two main types of CD4(+) Tregs: forkhead box protein 3 (FOXP3)(+) Tregs and interleukin-10-producing type 1 Tregs (Tr1 cells). The aim of this review is to highlight current information on the characteristics of human FOXP3(+) Tregs and Tr1 cells that make them an attractive therapeutic target. We discuss the progress and limitations that must be overcome to develop methods to enhance Tregs in vivo, expand or induce them in vitro for adoptive transfer, and/or inhibit their function in vivo. Although many technical and theoretical challenges remain, the next decade will see the first clinical trials testing whether Treg-based therapies are effective in humans.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/terapia , Linfocitos T CD4-Positivos/inmunología , Factores de Transcripción Forkhead/inmunología , Inmunoterapia Adoptiva , Neoplasias/inmunología , Neoplasias/terapia , Linfocitos T Reguladores/inmunología , Animales , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Enfermedad Injerto contra Huésped/prevención & control , Humanos , Tolerancia Inmunológica , Factores Inmunológicos/uso terapéutico , Activación de Linfocitos , RatonesRESUMEN
Forkhead box P3 (FOXP3) is considered a specific marker for CD4(+)CD25(+) regulatory T (Treg) cells, but increasing evidence suggests that human CD4(+)CD25(-) effector T (Teff) cells can transiently express FOXP3 upon activation. We demonstrate that the signal transducer and activator of transcription 5 (STAT5)-signaling cytokines, IL-2, IL-15 and to a lesser extent IL-7, induce FOXP3 up-regulation in vitro in activated human Teff cells. In contrast, cytokines which do not activate STAT5, such as IL-4 or transforming growth factor-beta alone, do not directly induce FOXP3 expression in activated Teff cells. Moreover, expression of a constitutively active form of STAT5a is sufficient to induce FOXP3 expression in Teff cells. Expression of FOXP3 in activated Teff cells requires both TCR-mediated activation and endogenous IL-2, but is not dependent on cell division and does not induce suppressive function. The presence of STAT5-activating cytokines is also required to maintain high FOXP3 expression and suppressive activity of Treg cells in vitro. These data indicate that activation of STAT5 sustains FOXP3 expression in both Treg and Teff cells and contribute to our understanding of how cytokines affect the expression of FOXP3.