RESUMEN
Developing a vaccine to prevent congenital cytomegalovirus (CMV) infection and newborn disability requires an understanding of infection incidence. In a prospective cohort study of 363 adolescent girls (NCT01691820), CMV serostatus, primary infection, and secondary infection were determined in blood and urine samples collected at enrollment and every 4 months for 3 years. Baseline CMV seroprevalence was 58%. Primary infection occurred in 14.8% of seronegative girls. Among seropositive girls, 5.9% had ≥4-fold increase in anti-CMV antibody, and 23.9% shed CMV DNA in urine. Our findings provide insights on infection epidemiology and highlight the need for more standardized markers of secondary infection.
Cytomegalovirus (CMV) can be passed from a woman to her unborn baby during pregnancy, which can result in disabilities in the baby. This can happen after a first infection with the virus during pregnancy, after a subsequent infection with a different strain ("reinfection"), or after "reactivation", which means that a virus present from a previous infection becomes active again. Vaccinating adolescent girls against CMV may be a future strategy to help prevent CMV infection during pregnancy. To provide information to design trials evaluating a CMV vaccine, it is important to know how common primary/secondary CMV infection is in adolescent girls and if this can be measured with available tools. We followed adolescent girls living in Finland, Mexico or the United States for three years. At study start, 58% of these girls showed evidence of previous CMV infection. During the three-year follow-up, a first CMV infection occurred in 15% of girls, and reinfection or reactivation in 6% to 24% of girls (depending on the method used). The obtained estimates of CMV infection rates in adolescent girls provide valuable information for future studies to evaluate CMV vaccines, but standardized markers for secondary infection are needed.
Asunto(s)
Coinfección , Infecciones por Citomegalovirus , Adolescente , Femenino , Humanos , Anticuerpos Antivirales , Citomegalovirus , Incidencia , Estudios Prospectivos , Estudios SeroepidemiológicosRESUMEN
We present a patient with suspected Proteus Syndrome, an overgrowth disorder associated with AKT1c.49G>A mutation. NGS analysis detected PIK3CAc.3140A>G mutation in the patient's affected tissue allowing for PROS (PIK3CA-related overgrowth spectrum) diagnosis. The overlapping clinical features in overgrowth disorders highlight the importance of molecular testing for a correct diagnosis.
RESUMEN
Adoptive cell therapy of solid tumors with reprogrammed T cells can be considered the "next generation" of cancer hallmarks. CAR-T cells fail to be as effective as in liquid tumors for the inability to reach and survive in the microenvironment surrounding the neoplastic foci. The intricate net of cross-interactions occurring between tumor components, stromal and immune cells leads to an ineffective anergic status favoring the evasion from the host's defenses. Our goal is hereby to trace the road imposed by solid tumors to CAR-T cells, highlighting pitfalls and strategies to be developed and refined to possibly overcome these hurdles.
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Terapia Genética/métodos , Inmunoterapia Adoptiva/métodos , Neoplasias/terapia , Receptores Quiméricos de Antígenos/inmunología , Animales , Edición Génica , Técnicas de Transferencia de Gen , Humanos , Neoplasias/genética , Neoplasias/inmunología , Neoplasias/patología , Receptores Quiméricos de Antígenos/genética , Microambiente TumoralRESUMEN
Molecular data concerning the involvement of roots in the genetic pathways regulating floral transition are lacking. In this study, we performed global analyses of the root transcriptome in Arabidopsis in order to identify flowering time genes that are expressed in the roots and genes that are differentially expressed in the roots during the induction of flowering. Data mining of public microarray experiments uncovered that about 200 genes whose mutations are reported to alter flowering time are expressed in the roots (i.e. were detected in more than 50% of the microarrays). However, only a few flowering integrator genes passed the analysis cutoff. Comparison of root transcriptome in short days and during synchronized induction of flowering by a single 22-h long day revealed that 595 genes were differentially expressed. Enrichment analyses of differentially expressed genes in root tissues, gene ontology categories, and cis-regulatory elements converged towards sugar signaling. We concluded that roots are integrated in systemic signaling, whereby carbon supply coordinates growth at the whole plant level during the induction of flowering. This coordination could involve the root circadian clock and cytokinin biosynthesis as a feed forward loop towards the shoot.
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Proteínas de Arabidopsis/biosíntesis , Arabidopsis/genética , Flores/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Redes Reguladoras de Genes , Genes de Plantas , Raíces de Plantas/fisiología , Arabidopsis/anatomía & histología , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Minería de Datos , Bases de Datos Genéticas , Conjuntos de Datos como Asunto , Metabolismo Energético/genética , Estudios de Asociación Genética , Mutación , Fotoperiodo , Hojas de la Planta/metabolismo , Raíces de Plantas/metabolismo , Regiones Promotoras Genéticas/genética , ARN de Planta/biosíntesis , ARN de Planta/genética , Elementos Reguladores de la Transcripción , Análisis de Matrices Tisulares , TranscriptomaRESUMEN
BACKGROUND AIMS: Chimeric antigen receptors (CARs) designed for adoptive immunotherapy need to achieve two functions: antigen recognition and triggering of the lytic machinery of reprogrammed effector cells. Cytotoxic T cells have been engineered with FcγRIII (CD16) chimeric molecules to be redirected against malignant cells by monoclonal antibodies (mAbs). These cells have been proven to mediate granule-dependent cellular cytotoxicity, but it is not clear whether they can also kill malignant cells by a granule-independent mechanism of cell cytotoxicity. METHODS: We engineered a CD16A-CAR equipped with the extracellular CD16A, the hinge spacer and the transmembrane region of CD8, and the ζ-chain of the T-cell receptor/CD3 complex in tandem with the CD28 co-stimulatory signal transducer module. The CD16A-CAR was expressed and functionally tested in the MD45 cell line, a murine T-cell hybridoma with a defective granular exocytosis pathway but capable of killing target cells by a Fas ligand-mediated lysis. RESULTS: Our results indicate that in vitro cross-linking of CD16A-CAR on MD45 cells by the Fc fragment of mAb opsonized tumor cells induced interleukin-2 release and granule-independent cellular cytotoxicity. CONCLUSIONS: We conclude that strategies aimed to implement the therapeutic functions of mAbs used in the clinic with T-dependent immune responses driven by engineered T cells expressing FcγR-CAR can boost the antitumor efficacy of mAbs used in the clinic.
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Anticuerpos Monoclonales/inmunología , Citotoxicidad Inmunológica/inmunología , Interleucina-2/metabolismo , Receptores de IgG/genética , Linfocitos T Citotóxicos/inmunología , Animales , Antígenos CD8/genética , Antígenos CD8/inmunología , Línea Celular , Proteína Ligando Fas/metabolismo , Proteínas Ligadas a GPI/biosíntesis , Proteínas Ligadas a GPI/genética , Humanos , Inmunoglobulina G/inmunología , Inmunoterapia Adoptiva/métodos , Ratones , Receptores de Antígenos/metabolismo , Receptores de IgG/biosíntesis , Receptores de IgG/inmunologíaRESUMEN
Root chicory (Cichorium intybus var. sativum) is a biennial crop, but is harvested to obtain root inulin at the end of the first growing season before flowering. However, cold temperatures may vernalize seeds or plantlets, leading to incidental early flowering, and hence understanding the molecular basis of vernalization is important. A MADS box sequence was isolated by RT-PCR and named FLC-LIKE1 (CiFL1) because of its phylogenetic positioning within the same clade as the floral repressor Arabidopsis FLOWERING LOCUS C (AtFLC). Moreover, over-expression of CiFL1 in Arabidopsis caused late flowering and prevented up-regulation of the AtFLC target FLOWERING LOCUS T by photoperiod, suggesting functional conservation between root chicory and Arabidopsis. Like AtFLC in Arabidopsis, CiFL1 was repressed during vernalization of seeds or plantlets of chicory, but repression of CiFL1 was unstable when the post-vernalization temperature was favorable to flowering and when it de-vernalized the plants. This instability of CiFL1 repression may be linked to the bienniality of root chicory compared with the annual lifecycle of Arabidopsis. However, re-activation of AtFLC was also observed in Arabidopsis when a high temperature treatment was used straight after seed vernalization, eliminating the promotive effect of cold on flowering. Cold-induced down-regulation of a MADS box floral repressor and its re-activation by high temperature thus appear to be conserved features of the vernalization and de-vernalization responses in distant species.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiología , Cichorium intybus/fisiología , Proteínas de Dominio MADS/genética , Proteínas de Plantas/genética , Proteínas de Arabidopsis/genética , Cichorium intybus/genética , Clonación Molecular , Frío , Flores/genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Dominio MADS/metabolismo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Temperatura , Regulación hacia ArribaRESUMEN
Cytokinins are involved in many aspects of plant growth and development, and physiological evidence also indicates that they have a role in floral transition. In order to integrate these phytohormones into the current knowledge of genetically defined molecular pathways to flowering, we performed exogenous treatments of adult wild type and mutant Arabidopsis plants, and analysed the expression of candidate genes. We used a hydroponic system that enables synchronous growth and flowering of Arabidopsis, and allows the precise application of chemicals to the roots for defined periods of time. We show that the application of N6-benzylaminopurine (BAP) promotes flowering of plants grown in non-inductive short days. The response to cytokinin treatment does not require FLOWERING LOCUS T (FT), but activates its paralogue TWIN SISTER OF FT (TSF), as well as FD, which encodes a partner protein of TSF, and the downstream gene SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1). Treatment of selected mutants confirmed that TSF and SOC1 are necessary for the flowering response to BAP, whereas the activation cascade might partially act independently of FD. These experiments provide a mechanistic basis for the role of cytokinins in flowering, and demonstrate that the redundant genes FT and TSF are differently regulated by distinct floral-inducing signals.
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Proteínas de Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/fisiología , Citocininas/fisiología , Proteínas de Unión a Fosfatidiletanolamina/genética , Reguladores del Crecimiento de las Plantas/fisiología , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Secuencia de Bases , Compuestos de Bencilo/farmacología , Citocininas/farmacología , ADN de Plantas/genética , Flores/efectos de los fármacos , Flores/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas/efectos de los fármacos , Proteínas de Dominio MADS/genética , Mutación , Reguladores del Crecimiento de las Plantas/farmacología , Plantas Modificadas Genéticamente , Purinas/farmacología , Transducción de Señal , Factores de Transcripción/genética , Activación Transcripcional/efectos de los fármacosRESUMEN
Molecular genetic analyses in Arabidopsis disclosed a genetic pathway whereby flowering is induced by the photoperiod. This cascade is examined here within the time course of floral transition in the long-day (LD) plant Sinapis alba induced by a single photoperiodic cycle. In addition to previously available sequences, the cloning of CONSTANS (SaCO) and FLOWERING LOCUS T (SaFT) homologues allowed expression analyses to be performed to follow the flowering process step by step. A diurnal rhythm in SaCO expression in the leaves was observed and transcripts of SaFT were detected when light was given in phase with SaCO kinetics only. This occurred when day length was extended or when a short day was shifted towards a 'photophile phase'. The steady-state level of SaFT transcripts in the various physiological situations examined was found to correlate like a rheostat with floral induction strength. Kinetics of SaFT activation were also consistent with previous estimations of translocation of florigen out of leaves, which could actually occur after the inductive cycle. In response to one 22-h LD, initiation of floral meristems by the shoot apical meristem (SAM) started about 2 days after activation of SaFT and was marked by expression of APETALA1 (SaAP1). Meanwhile, LEAFY (SaLFY) was first up-regulated in leaf primordia and in the SAM. FRUITFULL (SaFUL) was later activated in the whole SAM but excluded from floral meristems. These patterns are integrated with previous observations concerning upregulation of SUPPRESSOR OF OVEREXPRESSION OF CO1 (SaSOC1) to provide a temporal and spatial map of floral transition in Sinapis.
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Flores/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Fotoperiodo , Sinapis/genética , Secuencia de Aminoácidos , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Clonación Molecular , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Flores/genética , Regulación del Desarrollo de la Expresión Génica , Genes de Plantas , Proteínas de Dominio MADS/genética , Proteínas de Dominio MADS/metabolismo , Datos de Secuencia Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismo , Alineación de Secuencia , Sinapis/crecimiento & desarrollo , Sinapis/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
* Of the Brassicaceae, Sinapis alba has been intensively studied as a physiological model of induction of flowering by a single long day (LD), while molecular-genetic analyses of Arabidopsis thaliana have disclosed complex interactions between pathways controlling flowering in response to different environmental cues, such as photoperiod and vernalization. The vernalization process in S. alba was therefore analysed here. * The coding sequence of S. alba SaFLC, which is orthologous to the A. thaliana floral repressor FLOWERING LOCUS C, was isolated and the transcript levels quantified in different conditions. * Two-week-old seedlings grown in noninductive short days (SDs) were vernalized for 1-6 wk. Down-regulation of SaFLC was already marked after 1 wk of cold but 2 wk was needed for a significant acceleration of flowering. Flower buds were initiated during vernalization. When vernalization was stopped after 1 wk, repression of SaFLC was not stable but a significant increase in plant responsiveness to 16-h LDs was observed when LDs followed immediately after the cold treatment. * These results suggest that vernalization does not only work when plants experience long exposure to cold during the winter: shorter cold periods might stimulate flowering of LD plants if they occur when photoperiod is increasing, such as in spring.
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Frío , Flores/fisiología , Fotoperiodo , Proteínas de Plantas/metabolismo , Proteínas Represoras/metabolismo , Sinapis/fisiología , Secuencia de Aminoácidos , Southern Blotting , ADN Complementario/aislamiento & purificación , ADN de Plantas/metabolismo , Regulación hacia Abajo , Flores/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Datos de Secuencia Molecular , Proteínas de Plantas/química , Proteínas de Plantas/genética , Sinapis/genética , Factores de TiempoRESUMEN
In Arabidopsis thaliana, vernalization promotes flowering by repressing the floral inhibitor FLOWERING LOCUS C (AtFLC). This repression is mediated through epigenetic modifications at the AtFLC locus, leading to gene silencing. Whether the well-known quantitative effect of vernalization is due to the degree of AtFLC repression and/or its stability after return to normal temperature conditions has not been clarified. Here, we examine this question in white mustard, Sinapis alba, taking advantage of our recent cloning of the AtFLC ortholog SaFLC.