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Copyright © 2021 Gonçalves et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license., Activation of immune cells in response to fungal infection involves the reprogramming of their cellular metabolism to support antimicrobial effector functions. Although metabolic pathways such as glycolysis are known to represent critical regulatory nodes in antifungal immunity, it remains undetermined whether these are differentially regulated at the interindividual level. In this study, we identify a key role for 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) in the immunometabolic responses to Aspergillus fumigatus. A genetic association study performed in 439 recipients of allogeneic hematopoietic stem cell transplantation (HSCT) and corresponding donors revealed that the donor, but not recipient, rs646564 variant in the PFKFB3 gene increased the risk of invasive pulmonary aspergillosis (IPA) after transplantation. The risk genotype impaired the expression of PFKFB3 by human macrophages in response to fungal infection, which was correlated with a defective activation of glycolysis and the ensuing antifungal effector functions. In patients with IPA, the risk genotype was associated with lower concentrations of cytokines in the bronchoalveolar lavage fluid samples. Collectively, these findings demonstrate the important contribution of genetic variation in PFKFB3 to the risk of IPA in patients undergoing HSCT and support its inclusion in prognostic tools to predict the risk of fungal infection in this clinical setting. IMPORTANCE The fungal pathogen Aspergillus fumigatus can cause severe and life-threatening forms of infection in immunocompromised patients. Activation of glycolysis is essential for innate immune cells to mount effective antifungal responses. In this study, we report the contribution of genetic variation in the key glycolytic activator 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) to the risk of invasive pulmonary aspergillosis (IPA) after allogeneic hematopoietic stem cell transplantation. The PFKFB3 genotype associated with increased risk of infection was correlated with an impairment of the antifungal effector functions of macrophages in vitro and in patients with IPA. This work highlights the clinical relevance of genetic variation in PFKFB3 to the risk of IPA and supports its integration in risk stratification and preemptive measures for patients at high risk of IPA., This work was supported by the Fundação para a Ciência e Tecnologia (FCT) (PTDC/SAU-SER/29635/2017, PTDC/MED-GEN/28778/2017, UIDB/50026/2020, and UIDP/50026/2020), the Northern Portugal Regional Operational Program (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) (NORTE-01-0145-FEDER-000039), the Institut Mérieux (Mérieux Research Grant 2017), the European Society of Clinical Microbiology and Infectious Diseases (ESCMID Research Grant 2017), the European Union’s Horizon 2020 research and innovation program under grant agreement no. 847507, and the “la Caixa” Foundation (ID 100010434) and FCT under the agreement LCF/PR/HR17/52190003. Individual support was provided by FCT (SFRH/BD/136814/2018 to S.M.G., PD/BD/137680/2018 to D.A., CEECIND/04058/2018 to C.C., and CEECIND/03628/2017 to A.C.). M.G.N. was supported by an ERC Advanced Grant and a Spinoza Grant of the Netherlands Organization for Scientific Research.

En tiempos de encierro y cuarentena, la radio tiene un gran espacio entre los consumos culturales. Sin poder salir de casa, con la computadora, el televisor y el celular a mano todo el día, la radio sigue encendida. ¿Por qué? ¿Qué es lo que hace que en tiempos de consumo de información y entretenimiento en hogares, dedónde no se puede (ni debe) salir, una voz que nos habla a través de un parlante que toma la señal del aire o de un reproductor de streaming sea atractivo e interesante? Primero los datos. Las principales corporaciones de radio en Reino Unido indican que sus oyentes en internet crecieron entre un 15% y un 18%. Ivoox, plataforma de radio en internet española, registra una caída en la escucha de podcasts pero un aumento del 20% en las conexiones a los streamings de las radios, en especial las informativas. Deezer, la plataforma musical, muestra lo mismo: la radio le gana al podcast y la música en tiempos de coronavirus. En Holanda, el tiempo de escucha de radio y la cantidad de oyentes marcan aumentos del 11% y el 7%, respectivamente. En Argentina, las radiostienen picos máximos históricos de conexión a sus reproductores. Kantar muestra que un 24% de los argentinos escucha más radio en internet., Facultad de Periodismo y Comunicación Social

Las radios y televisoras comunitarias en la Argentina cuentan con una larga y diversa trayectoria que se remonta a cuatro décadas atrás, y se encuentran en todo el territorio del país. A pesar de su relevancia, no existen aún datos completos y fiables sobre la cantidad y características de emisoras activas en el Estado, las universidades, ni las redes que las nuclean. En un trabajo inédito, este artículo releva y analiza los medios comunitarios, populares y alternativos actualmente operativos. Se basa en una exhaustiva investigación empírica de dos años que recolectó, depuró y sistematizó información proveniente de distintos tipos de fuentes. Se argumenta que, a partir de 2008, cuando se inicia el debate público del proyecto de Ley Audiovisual, se produjo un exponencial crecimiento de la cantidad de radios y televisoras del sector social en el país. Esta correlación y estudios previos permiten inferir que las políticas de legalización y promoción establecidas por esa norma tienen una incidencia relevante en ese resultado. Community radios and TV stations in Argentina have a long and diverse history that goes back four decades, and they are found throughout the country. Despite its relevance, there are still no complete and reliable data on the amount of active media and their characteristics in the State, universities, or the networks that group them. In an singular work, this article analyzes the community, popular and alternative media currently operative. It is based on an exhaustive two-year empirical research that collected, refined and systematized information from different types of sources. It is argued that, from 2008, when the public debate of the Audiovisual Law project began, there was an exponential growth in the number of radios and television stations in the social sector in the country. This correlation and previous studies allow us to infer that the legalization and promotion policies established by that norm have a relevant impact on this result. Fil: Segura, Maria Soledad. Universidad Nacional de Córdoba. Facultad de Filosofía y Humanidades. Centro de Investigaciones María Saleme Burnichón; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Linares, Alejandro Felix. Universidad de Buenos Aires. Facultad de Ciencias Sociales. Instituto de Estudios de América Latina y el Caribe; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Espada, Agustín Eduardo. Universidad Nacional de Quilmes. Departamento de Ciencias Sociales. Instituto de Estudios Sociales de la Ciencia y la Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Longo, Verónica Beatriz. Universidad Nacional de San Luis; Argentina Fil: Hidalgo, Ana Laura. Universidad Nacional de San Luis. Facultad de Ciencias Humanas. Departamento de Comunicación; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; Argentina Fil: Traversaro, Natalia Gabriela. Universidad Nacional de Córdoba; Argentina Fil: Vinelli, Natalia Andrea. Universidad de Buenos Aires; Argentina

El presente ensayo propone un análisis descriptivo de un instrumento de regulación de la PO en la Ciudad Autónoma de Buenos Aires (CABA), Argentina, novedosa por su objetivo: promover e impulsar la actividad de los medios locales. De esta forma, la Ley N° 2.587 “de Medios Vecinales de Comunicación Social” resulta relevante por generar un instrumento de fomento para medios con y sin fines de lucro en el distrito. El objetivo de la propuesta consiste en identificar las características principales de esta regulación y las de su aplicación. Se encuadra en estudios de la economía política crítica y las políticas públicas.Así, en primer lugar, se describirá los rasgos constitutivos del ecosistema de medios en Argentina. Luego, se describirá el instrumento de regulación enfocado en el análisis. A continuación, se dará cuenta de su aplicación y los efectos de los cambios recientes. Para completar el recorrido con algunas conclusiones sobre el objeto.

espanolLa diversidad cultural resulta imprescindible a la hora de poder representar la multiplicidad de formas mediante las cuales se expresan los diferentes grupos de una sociedad. El presente trabajo aborda un analisis cuantitativo de la musica programada en las seis radioemisoras mas escuchadas del Area Metropolitana de Buenos Aires, Argentina, entre los anos 2011 y 2017. Los parametros escogidos para mensurar el nivel de diversidad musical existente en la programacion de dichas radios son la cantidad de canciones emitidas, la cantidad de titulos de canciones, numero de artistas y el origen de los mismos. Por medio de esta ultima variable, se pretende demostrar si se cumple o no con la ultima regulacion de servicios de comunicacion audiovisual en la Argentina, la cual fija una cuota de musica nacional y musica nacional independiente. Por ultimo, se exhibe el grado de concentracion de productores fonograficos de las canciones y artistas emitidos. EnglishCultural diversity is essential when it comes to represent the multiplicity of ways in which different groups of a society are expressed. The present work deals with a quantitative analysis of the music programmed in the six radio stations most listened in the Metropolitan Area of Buenos Aires, Argentina, between 2011 and 2017. The parameters chosen to measure the level of musical diversity existing in the programming of those radios are the quantity of songs emitted, the amount of titles of songs, number of artists and the origin of the same ones. By means of this last variable, it is tried to demonstrate the level of compliance with the last regulatory process of audiovisual communication services in Argentina, which fixes a national music and indie national music share. Finally, the degree of concentration of phonographic producers of the songs and artists cast is shown.

Media ownership in Argentina is a common object of study in investigations that analyze this system’s economic–political history. The present article aims to contribute to this tradition, offering a description and analysis of the structure of the radio market in the Autonomous City of Buenos Aires since Argentina’s return to democracy (1983–2016). Our suggested itinerary covers the trading processes undertaken by broadcasting licensee associations in relation to the political changes and governments that succeeded each other during our chosen interval. The article begins with a description of the political and media situation in 1983. In later sections, it chronicles the relationship between politics, communication policies, and the radio system’s structure. Our time frame opens with Argentina’s return to democracy in 1983 and closes, in 2016, with Mauricio Macri’s first year as the country’s president. This span of thirty-odd years allows us to look at different governments, economic models, technological developments, and at the media market in general. The goal of this article is to interpret the behavior of capital in the Buenos Aires radio market from 1983 to 2016, and understand how this relates to what happened in the political and economic spheres. We seek to find relationships that either demonstrate or refute the influence of historical processes in the current state of this cultural industry.

Clinical disease caused by BK virus reactivation is a frequent complication of allogeneic hematopoietic cell transplantation (HCT). Because of the lack of effective antiviral agents, BK virus–specific T cells are emerging as a potential therapy for BK virus disease, but the immune response to BK virus after allogeneic HCT has not been well characterized. Our study describes reconstitution of BK virus–specific T-cell immunity in 77 adult patients after HCT. All patients had urinary symptoms, and urine was tested for BK virus replication; 33 patients were positive for BK virus (cases), and 44 were negative (controls). In BK virus cases, the median time to first positive test was 75 days (range, 2-511). BK virus cases had lower CD4 T-cell counts 3 to 9 months after transplant, but CD8 T-cell counts were similar in cases and controls. BK virus–specific T cells were identified by cytokine flow cytometry in cryopreserved samples collected prospectively. BK virus–specific CD4 T cells producing T helper 1 (Th1) cytokines recovered quickly after HCT. BK virus–specific T cells were detected more frequently in patients with BK virus reactivation at most time points, and CD4 T cells producing Th1 cytokines were more frequent than BK virus–specific cytolytic CD8 T cells. Early detection of interferon-γ+ and cytolytic BK virus–specific CD4 T cells was associated with lower rates of hematuria among cases. Overall, our study describes recovery of BK virus–specific T cells after HCT and the distinct roles for BK virus–specific T cells in the development and resolution of clinical symptoms.

Este artículo propone un acercamiento explorativo por las estrategias de producción de contenidos y de comercialización de las radios de AM y FM más escuchadas de Buenos Aires, Argentina, para sus sitios webs. De esta manera, y como objetivo general del trabajo, se busca indagar y descubrir innovaciones o experiencias en la generación-producción de contenidos radiofónicos digitales así como en su comercialización o venta publicitaria.El estudio de los cambios que propone la convergencia tecnológica y mediática para las empresas de medios de comunicación implica la realización de distintos trabajos simultáneos. La observación y el análisis de los contenidos es sólo una parte que se complementa, en este trabajo, con la realización de entrevistas a productores, gerentes y directores de radios. En especial de sus secciones o departamentos digitales.De esta forma, uno de los principales hallazgos de este trabajo consiste en la falta de estrategias definidas, la ausencia de producciones radiofónicas específicas para la web y una búsqueda de mayor complementariedad con los contenidos tradicionales a través de textos y videos. En este marco, lo incierto del comportamiento del mercado publicitario tradicional atenta contra la toma de iniciativas por parte de las gerencias., This article proposes an exploratory approach to the content production and marketing strategies of the most listened AM and FM radios in Buenos Aires, Argentina, for their websites. In this way, and as a general objective of the work, we seek to investigate and discover innovations or experiences in the generation-production of digital radio content as well as in its commercialization or advertising sale.The study of the changes proposed by the technological and media convergence for media companies implies the realization of different simultaneous works. The observation and analysis of the contents is only a part that is complemented, in this work, with the realization of interviews to producers, managers and radio directors. Especially its digital sections or departments.In this way, one of the main findings of this work consists in the lack of defined strategies, the absence of specific radio productions for the web and a search for greater complementarity with traditional contents through texts and videos. In this context, the uncertainty of the behavior of the traditional advertising market is against the taking of initiatives by management.

Key Clinical Message Hemophagocytic lymphohistiocytosis (HLH) should be considered in the differential diagnosis of adult patients with white matter disease. Brain involvement can be life‐threatening and should prompt aggressive therapy. Even after HLH remission, the possibility of subsequent deterioration due to emergence of an aggressive intravascular lymphoma is highlighted here.

Desde 2004, por primera vez en la historia de la radiodifusión de la región, una decena de países latinoamericanos reconocieron a las radios y televisiones comunitarias, alternativas y populares como prestadoras legales de servicios de comunicación audiovisual. Este cambio en las regulaciones y políticas estatales se dio junto con una modificación tecnológica por la expansión de las posibilidades de prestación de servicios convergentes. Ante las nuevas condiciones, las estrategias de estas emisoras cambiaron. Lo que sucedió con sus públicos en este nuevo contexto es un tema que ha recibido todavía escasa atención académica. Se procura responder los siguientes interrogantes: ¿Cuál es el público objetivo que construyen los medios comunitarios en sus estrategias comunicacionales? ¿Qué características tienen sus públicos reales? ¿Cuáles son sus niveles de participación? ¿Qué significaciones construyen alrededor de estas emisoras? Se presentan los resultados de 10 entrevistas a integrantes de 7 radios y 3 televisiones de cuatro provincias de tres regiones distintas de Argentina, 37 encuestas cerradas de consumo de medios entre sus televidentes y oyentes, y 6 focus groups con ellos/as. Since 2004, for the first time in the history of radio broadcasting in the region, a dozen Latin American countries have recognized alternative and popular community radios and television stations as legal providers of audiovisual communication services. This change in the regulations and state policies occurred along with a technological modification due to the expansion of the possibilities of providing convergent services. Given the new conditions, the strategies of these TV and radio stations changed. What happened to their publics in this new context is a topic that has still received little academic attention. We answer the following questions: Which are the publics that community media expect and construct in their communication strategies? Which characteristics do their real publics have? What are their participation levels? What meanings do they propose around these stations? We present the results of 10 interviews with members of the 7 radio and 3 TV stations of four states of three different regions of Argentina, 37 closed surveys of media consumption among their viewers and listeners, and 6 focus groups with them.

Chronic graft-versus-host disease (cGvHD) remains a major complication of allogeneic stem cell transplantation requiring novel therapies. CD146 and CCR5 are expressed by activated T cells and associated with increased T cell migration capacity and Th17 polarization. We performed a multiparametric flow cytometry analysis in a cohort of 40 HSCT patients together with a cGvHD murine model to understand the role of CD146-expressing subsets. We observed an increased frequency of CD146+ CD4 T cells in the 20 patients with active cGvHD with enhanced RORγt expression. This Th17-prone subset was enriched for cells coexpressing CD146 and CCR5 that harbor mixed Th1/Th17 features and were more frequent in cGvHD patients. Utilizing a murine cGvHD model with bronchiolitis obliterans (BO), we observed that donor T cells from CD146-deficient mice versus those from WT mice caused significantly reduced pulmonary cGvHD. Reduced cGvHD was not the result of failed germinal center B cell or T follicular helper cell generation. Instead, CD146-deficient T cells had significantly lower pulmonary macrophage infiltration and T cell CCR5, IL-17, and IFN-γ coexpression, suggesting defective pulmonary end-organ effector mechanisms. We, thus, evaluated the effect of TMP778, a small-molecule RORγt activity inhibitor. TMP778 markedly alleviated cGvHD in murine models similarly to agents targeting the Th17 pathway, such as STAT3 inhibitor or IL-17–blocking antibody. Our data suggest CD146-expressing T cells as a cGvHD biomarker and suggest that targeting the Th17 pathway may represent a promising therapy for cGvHD.

En Latinoamérica, en particular en Argentina, la gestión de los servicios públicos de radiodifusión ha variado al ritmo de los cambios de gobierno. Sin embargo, la concepción de los servicios públicos de radiodifusión como entes y medios gubernamentales se sostuvo de manera constante a lo largo del tiempo. Este trabajo se propone estudiar la gestión de este tipo de servicios y su definición conceptual en un escenario concreto dado por el período de gobiernos kirchneristas (2003-2015). Para esto se toma como objeto de estudio a Radio Nacional. De esta manera, el objetivo consiste en demostrar de qué forma se manifiestan los cambios políticos en el desarrollo del servicio público de radiodifusión más antiguo del país. In Latin America, and therefore in Argentina, the management of public broadcasting service has changed to heat and pace of change of government. This implies that, beyond changes in management, the concept of "public broadcasting service" as governmental entities and media held steadily over time. This paper intends to approach the management of these services and their conceptual definition in a particular setting given by the period of government of the kirchneristas (2003- 2015). To addres this matter, the National Radio is taken as the object of study. Thus, the objective is to demonstrate how politics and policies changes are manifested in the development of the country's oldest public broadcasting service. Fil: Espada, Agustín Eduardo. Universidad Nacional de Quilmes. Departamento de Ciencias Sociales. Industrias Culturales y Espacio Público; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Fil: Espada, Agustín. Universidad Nacional de Quilmes; Argentina. Fil: Marino, Santiago. Universidad Nacional de Quilmes; Argentina. Espada, A. y Marino, S. (2018). Publicidad Oficial en Argentina: propuesta metodológica y análisis crítico. Divulgatio. Perfiles académicos de posgrado, 3(7), 112-133. Desde la Economía Política de la Comunicación (EPC) es posible estudiar los procesos de producción, distribución y consumo de bienes y servicios culturales (Zallo, 1988 y 2009). Si el foco se pone en la relación entre el Estado y los agentes del mercado se agregan elementos propios de los estudios de las políticas públicas de comunicación. En el actual contexto de transformación de procesos y economía, este trabajo propone un abordaje específico sobre Publicidad Oficial (PO), un aspecto clave de la relación entre los estados y el sistema de medios. De este modo, en las próximas páginas se parte de una concepción entiende a la PO como una de las herramientas con las que el Estado cuenta para dar a conocer sus políticas, actos oficiales y servicios. Como así también en tanto que un mecanismo para administrar la relación con las empresas privadas de medios de comunicación. Desde este punto de entrada conceptual, se combina el análisis teórico y práctico de regulaciones sobre la materia y desgloses de gastos del Poder Ejecutivo Nacional (PEN) argentino durante los años 2015 y 2016. Esto implica, entonces, analizar el último año de la presidencia de Cristina Fernández de Kirchner y el primero de Mauricio Macri, respectivamente. Se intenta de este modo realizar un aporte a un ámbito con cierta vacancia en el estudio de las políticas públicas de comunicación. Se subraya la intención de establecer un conjunto de principios y una metodología para el desglose de los gastos de un organismo o una esfera del Estado en un momento y lugar determinados. From the Political Economy of Communication (EPC) it is possible to study the processes of production, distribution and consumption of cultural goods and services. If the focus is placed on the relationship between the State and market agents, elements of the studies of public communication policies are added. In the current context of transformation of processes and economies, studies emerge on how the State administers, executes and regulates the media system. A significant approach is the one that analyzes the Official Publicity, a key aspect. In this work Official Publicity (PO) is understood as one of the tools with which the State counts on publicizing its policies, official acts and services. From this conceptual entry point, the work combines the theoretical and practical analysis of regulations on the subject and expenditure breakdowns of the Argentine National Executive Power during the years 2015 and 2016, last of the presidency of Cristina Fernández de Kirchner and first of Mauricio Macri respectively. The objective of the work is to make a contribution to a field with a certain vacancy in the study of public communication policies. It underlines the intention to establish a set of principles and methodology for the breakdown of the expenses of a body or a sphere of the State at a particular time and place.

El sistema de medios de comunicación y la producción de información y cultura se transforma a nivel mundial. Las fuerzas combinadas de la digitalización y la masificación de internet como plataforma de acceso y distribución de contenidos modifican las reglas de juego. En este contexto, la radio participa con la aparición de nuevos actores, formatos y modelos que amplían el campo de “lo radiofónico”. El ecosistema radiofónico online es el mar en el que estas novedades se desarrollan, combinan y transforman. Allí surgen los podcast como nuevos formatos de producción y distribución de contenidos sonoros con características radiofónicas. El movimiento podcaster está formado por una amplia variedad de participantes: pequeños, medianos, independientes, corporativos, fugaces, históricos. Este artículo propone la descripción de cuatro casos caracterizados como “redes de podcast”. Estos proyectos fueron diseñados para y subsisten con la producción de varios programas y aparecen así como emprendimientos pensados por y para la producción de podcast. La pregunta que se hace aquí es puramente descriptiva y exploratoria: ¿cómo producen, distribuyen y comercializan sus contenidos las redes de podcast en Argentina? Para responderla se realizan un trabajo cualitativo de entrevistas y análisis de contenidos y otro cuantitativo para los niveles de comparación entre los volúmenes y formatos de cada una de las redes., The media system and the production of information and culture is transformed worldwide. The combined forces of digitization and the massification of the internet as a platform for access and distribution of content modify the rules of the game. In this context, the radio participates with the appearance of new actors, formats and models that expand the field of "the radio." The online radio ecosystem is the sea in which these novelties are developed, combined and transformed. Podcasts emerge there as new production formats and distribution of sound content with radio features. The podcaster movement is made up of a wide variety of participants: small, medium, independent, corporate, fleeting, historical. This article proposes the description of four cases characterized as "podcast networks". These projects were designed for and subsist with the production of several programs and appear as well as ventures designed by and for the production of podcasts. The question that is asked here is purely descriptive and exploratory: how do podcast networks in Argentina produce, distribute and market their content? To answer it, a qualitative work of interviews and content analysis is carried out and a quantitative one for the levels of comparison between the volumes and formats of each of the networks., Facultad de Periodismo y Comunicación Social

The radio market in Argentina responds to global trends and has its own characteristics and actors too. Vorterix Rock is a multiplatform media emerged in 2012 that presents new answers to the structural conditions of argentine media system. In this article, the artistic and the commercial models are both analyzed to classify Vorterix's business model in accordance with either the traditional strategies that come from radio as a cultural industry or those from digital media. In addition, questions about the future of this business model and radio as a communication media are presented.

Introduction: BK virus (BKV) seropositivity is highly prevalent (80-90%) in healthy adults, but BKV rarely causes significant clinical disease. However, immunologic control of BKV is often compromised after allogeneic hematopoietic stem cell transplantation (HSCT) and BKV reactivation can cause clinical disease, including hemorrhagic cystitis, in 5-68% of patients in the first year after HSCT. Immune reconstitution of BKV-specific T cells after HSCT has not been well defined, and the role of BKV-specific T cells in response to clinical infection has not been studied. Methods: From the DFCI 2010-2011 HSCT cohort (Rorije et al, BBMT 2014), 33 adult patients were selected who had urinary symptoms, were tested for urine BKV by PCR, and were diagnosed with BKV disease (cases; n=16) or did not have BKV reactivation (controls; n=17). Cases and controls were matched for cyclophosphamide use, acute GvHD and stem cell source. BKV-specific T cells were analyzed in 180 prospectively cryopreserved peripheral blood mononuclear cells (PBMC) samples (average 5.5/patient) across several timepoints (pre-HSCT and 1, 3, 6, 9, 12, 18 and 24 months post-HSCT) using cytokine flow cytometry (CFC). PBMC were stimulated for 6 hours with overlapping 15-mer peptides derived from BKV LT and VP1 proteins in the presence of brefeldin A, monensin and anti-CD28/49d. Results were compared with no-pepmix negative controls and Staphylococcal enterotoxin B positive controls. After stimulation, cells were stained with fluorochrome-conjugated antibodies specific for CD3, CD4, CD8, CD45RA, CCR7, CD57, CD107a, IFNγ, TNFα, IL-2, perforin (PERF) and granzyme B (GZMB). Laboratory assessments were blinded to clinical disease status. Results: Median age of the cohort was 51 years (IQR; 39-61); 79% were male and acute GvHD occurred in 64%. Conditioning included cyclophosphamide in 46% and ATG in 24%; cell source was peripheral blood stem cells in 81%, cord blood in 15% and bone marrow in 3%. Median time from HSCT to BKV testing was 60 days (23-181); median time to first positive test was 79 days (26-196) in BKV+ cases. Median duration of symptoms was 53 days (11-92) in cases, compared to 5 days in controls. The percentage of patients with BKV-specific T cells across timepoints is summarized in Figure 1. The frequency of BKV-specific degranulating (CD107a) T cells peaked at 6mo, where they could be detected in 37.5% and 58.3% of patients by reactivity of CD8+ and CD4+ T cells, respectively. Detection of BKV-specific T cells producing any cytokines (cytokine+) peaked at 9 mo (CD8+ in 19% and CD4+ in 38.1% of patients). BKV-specific IFNγ+, IL-2+, and TNFα+ T cells all peaked at 12mo after HSCT. Subsequently, detection of BKV-specific CD8+ T cells declined sharply while BKV-specific CD4+ T cells were maintained at higher levels 24 months after HSCT. Comparing BKV-specific immune responses in cases and controls, IFNγ+ CD8+ cells reconstituted faster in patients with clinical disease (first detected at 3 months vs. 9 mo), and were present in more patients at 9 months: 41.7% vs. 11.1%. A similar trend was noted for IL-2+ CD8+, IFNγ+ CD4+, and IL-2+ CD4+ T cells (Figure 2). BKV-specific CD8+ T cells were predominately central memory (CM) and terminally differentiated effector memory and their cytokine profile increased gradually after transplant; 19% produced at least two cytokines at 1 mo, compared to 71% at 12 mo. BKV-specific CD4+ T cells were predominately CM and effector memory and their cytokine profile also increased after transplant; 3.5% produced at least two cytokines at 1 mo, compared to 35% at 12 mo and 69.5% at 24 mo. PERF and/or GZMB were detected in 45-74% of BKV-specific CD107a CD8+ cells. CD107a CD8+ cells also produced IFNγ. Conclusion: BKV-specific T cells gradually reconstitute in the first 9 months after allogeneic HSCT. 30-40% of patients with clinical symptoms and 50-60% of patients with confirmed BKV disease had detectable BKV-specific T cells but BKV-specific immunity recovers more rapidly in patients with BKV disease. As CD8 and CD4 BKV-specific immunity recovers, T cells gradually become more functional with a memory phenotype. Our cohort is currently being expanded to assess the correlation between the development and expansion of BKV-specific T cells and clinical outcomes. Disclosures Koreth: kadmon corp: Membership on an entity's Board of Directors or advisory committees; takeda pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; amgen inc: Consultancy; LLS: Research Funding; prometheus labs inc: Research Funding; millennium pharmaceuticals: Research Funding. Armand:Otsuka: Research Funding; BMS: Consultancy, Research Funding; Infinity: Consultancy; Sequenta: Research Funding; Roche: Research Funding; Merck & Co., Inc.: Consultancy, Research Funding; Sigma Tau: Research Funding; Tensha: Research Funding. Soiffer:GentiumSpA/Jazz Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees. Ritz:Kiadis: Membership on an entity's Board of Directors or advisory committees.

Chronic Graft versus Host disease (cGVHD) is a major limiting factor for the success of allo-HSCT. In this prospective study we aimed to evaluate the association between the kinetics of Regulatory T cells (TREG) and Conventional CD4+T cells (TCON) reconstitution and the emergence of cGVHD. We performed a detailed phenotypic analysis by multiparametric flow cytometry using fresh blood from 39 patients undergoing unrelated donor HSCT after a reduced intensity conditioning regimen containing ATG over a 2 year period, representing a total of 213 samples analyzed. GVHD prophylaxis consisted of cyclosporine plus mycophenolate mofetil. 11 patients were excluded due to disease relapse and/or death due to infection or acute GVHD in the first 9 months post-HSCT. We observed indiscriminately low numbers of TREG (CD4+CD127lowFoxp3+CD25bright) until mo 6 after HSCT and reduced TREG numbers in patients developing cGVHD (GVHD+) versus those who did not (GVHD-) (p=0.02 at mo 9). We further studied the dynamics of TREG subset reconstitution. CM TREG (CD45RA-CD62L+) was the predominant population in both patient groups. EM (CD45RA-CD62L-) was the second most abundant TREG subset. CM and EM TREG numbers were similar between patient groups. EMRA (CD45RA+CD62L-) TREG remained very low throughout the follow-up but were significantly increased at mo 9 in GVHD+ (p= 0.03). Interestingly, Naïve TREG (CD45RA+CD62L+CD95-) started to emerge at mo 9 and were significantly reduced in GVHD+ patients at mo 12 (0.71 vs 0.14 cells/µl; p=0.02) The Stem Cell Memory subset (SCM), identified as CD45RA+CD62L+CD95+, is thought to be self renewing and multipotent, being able to differentiate into CM, EM and TEMRA memory subsets. While in GVHD- SCM TREG started to emerge at mo 9, this subset remained low in GVHD+. Statistically significant differences were observed at mo 18 (0.91 vs 0.15 cells/µl; p=0.02). To ascertain the role of thymic output in TREG reconstitution we quantified CD31+ naïve TREG. Recent Thymic Emigrant cells (RTE) TREG were significantly reduced in GVHD+ at mo 12 (0.8 vs 0.16cells/µl; p=0.02) and at mo 18 (1.93 vs 0.28 cells/µl; p=0.02). In order to clarify whether both thymic output and peripheral expansion were contributing factors to decreased Naïve TREG, we quantified proliferation using Ki-67. TREG from GVHD+ patients proliferated less from months 2 to 18 reaching statistical significance at mo 9 and 12 (p= 0.003 and 0.02, respectively), suggesting that decreased TREG numbers are at least partly due to reduced peripheral expansion. Taken together, these observations suggest a compromised peripheral expansion and de novo generation of TREG through thymic output in cGVHD patients. Of note, susceptibility to apoptosis was not increased in TREG from GVHD+ patients, as Bcl-2 levels tended to be higher in relation to GVHD- patients. In the CD4+Foxp3- TCON population, we observed a clear predominance of EM in all patients. These cells, together with CM, are the first to appear at similar levels in both patient groups. EMRA emerged at higher numbers in GVHD+, although the results did not reach statistical significance. Naïve and RTE TCON started to emerge after mo 6 in GVHD-. Interestingly, these cells remained much lower in GVHD+ vs GVHD- patients throughout the follow-up, reaching statistical significance at mo 12 (p=0.03 for naïve; p=0.03 for RTE TCON). SCM TCON remained very low in all patients showing a tendency to be reduced in GVHD+ when compared to GVHD-. This difference reached statistical significance at mo 18 (6.85 vs 2.13 cells/µl; p=0.02). The small number of TREG and TCON subsets in GVHD+ patients were unlikely due to increased susceptibility to apoptosis, as assessed by Bcl-2 expression. The low numbers of cells within naïve and SCM TREG gates precluded the analysis of Bcl-2 expression in these subsets. Bcl-2 tended to be increased in total TREG CM and EM in GVHD+. In TCONs, Bcl-2 levels were similar in both patient groups. In summary, our data in patients developing cGVHD suggest that decreased thymic output and increased differentiation into terminally differentiated effector cells may have a negative impact on the number of naïve and SCM TCON and TREG. We speculate that the inability to generate and maintain the more immature TREG subsets may lead to decreased TREG numbers after 6 months post transplant, potentially resulting in decreased control of effector T cells contributing to the development of cGVHD. Disclosures Ritz: Kiadis: Membership on an entity's Board of Directors or advisory committees.

Introduction: Cytokines play important roles in the activation, proliferation, differentiation and survival of T cells. Previous studies have revealed that individual cytokines selectively activate different T cell populations and also function at specific stages of T cell differentiation. For example, IL-2 supports the development of CD4 regulatory T cells. IL-7 is required for naive conventional CD4 T cell (Tcon) homeostasis, whereas naive CD8 T cell homeostasis requires both IL-7 and IL-15. In contrast, IL-6 promotes Th17 T cell differentiation. The functions of each cytokine are partly defined by the differential expression of specific multi-unit receptors but the selective homeostatic effects of individual cytokines are still incompletely understood. Methods: We stimulated peripheral blood mononuclear cells from healthy donors with varying concentrations of IL-2, IL-7, IL-15 and IL-6 for 15 min in vitro. Single cell mass cytometry (CyTOF) with a panel of 33 markers was used to simultaneously examine signaling pathways activated by each cytokine in distinct T cell subsets. viSNE, a cytometry analysis tool, was used to visualize high-dimensional cytometry data on a two-dimensional map. Expression of pSTAT5 was used to monitor activation induced by IL-2, IL-7 and IL-15; pSTAT3 was used to monitor activation by IL-6. Results: In CD4 Tcon, relatively high concentrations of IL-2 (100-1000 IU/ml) are required to induce pSTAT5 (Figure 1). However even at high concentrations, IL-2 activation was selective for memory Tcon subsets. In contrast, IL-7 induced pSTAT5 at very low concentrations (1-10 IU/ml). Although all Tcon were affected, activation was more robust in memory than naive Tcon subsets at all IL-7 concentrations. IL-15 activation of pSTAT5 required at least 10 IU/ml and only memory Tcon subsets were activated even at high IL-15 concentrations. Whereas IL-2, IL-7 and IL-15 preferentially activated memory Tcon subsets, IL-6 selectively activated pSTAT3 in naive and central memory (CM) Tcon subsets at low concentrations (10 IU/ml). At high IL-6 concentrations (100-1000 IU/ml) effector memory (EM) Tcon were also activated. CD8 T cells (Figure 2) are relatively insensitive to IL-2, and only CM CD8 T cells are activated at high IL-2 concentrations (100 IU/ml). Although all CD8 T cell subsets were activated at very high IL-2 concentrations (1000 IU/ml), pSTAT5 activation remained most evident in CM CD8 T cells. Similar to Tcon, IL-7 induced pSTAT5 in CD8 T cells at very low IL-7 concentrations (1-10 IU/ml). However unlike Tcon, pSTAT5 activation was most prominent in naive and CM CD8 T cells. EM CD8 T cells were activated at higher IL-7 concentrations but TEMRA CD8 T cells were resistant to IL-7 stimulation. IL-15 induced pSTAT5 equally in all CD8 T cell subsets but relatively high concentrations (100-1000 IU/ml) were required. Similar to CD4 Tcon, IL-6 induced selective pSTAT3 activation in naive CD8 T cells. Activation of naive CD8 T cells was observed at low concentrations of IL-6 and both EM and TEMRA were resistant to very high IL-6 concentrations (100-1000 IU/ml). Conclusion: This detailed analysis of cytokine signaling has identified differential effects of IL-2, IL-7, IL-15 and IL-6 on different subsets of CD4 Tcon and CD8 T cells. Whereas CD4 Treg are activated at low IL-2 concentrations, CD4 Tcon and CD8 T cells are relatively resistant to IL-2. At high IL-2 concentrations, activation was most prominent for memory CD4 Tcon and CM CD8 T cells. In contrast, low concentrations of IL-7 are sufficient to activate both CD4 Tcon and CD8 T cells. Within these populations, memory Tcon and naive CD8 cells were preferentially activated at low IL-7 concentrations. Within the CD8 T cell population, IL-15 activated all subsets equally. Within CD4 Tcon, IL-15 preferentially activates memory subsets. IL-6 acts at low concentrations and primarily on naive cells in both CD4 Tcon and CD8 T cells. In all experiments, these effects do not require TCR antigen activation and therefore reflect the potency and differential activity of homeostatic signals supported by these cytokines. Importantly, high concentrations used for in vitro experiments are not likely achieved in vivo but may reflect toxicities of high dose exogenous cytokine therapies or cytokine release syndromes. In contrast, differential effects observed at low concentrations more likely reflect physiologic homeostatic effects of these cytokines in vivo. Disclosures No relevant conflicts of interest to declare.

Acute GVHD (aGVHD) remains a major cause of morbidity in patients who undergo allogeneic hematopoietic stem cell transplantation (HSCT). Although numerous studies have investigated potential immunologic parameters that could predict for aGVHD, there are no validated immune biomarkers presently used in clinical practice. We are currently performing a prospective analysis of immune reconstitution after allogeneic HSCT in different patient cohorts. The goal of the present study is to detect abnormalities of immune reconstitution associated with an inability to maintain T cell tolerance after allogeneic HSCT and to identify specific immunologic biomarkers predictive of aGVHD. We have thus far accrued 36 unrelated HSCT recipients with a median age of 48 years old (16 females; 20 males). The degree of HLA-matching at the allele level between patients and donors was: 10/10 (n=20), 9/10 (n=15), 8/10 (n=1). Graft source consisted of unmanipulated bone marrow (n=18) or peripheral blood stem cells (n=18). All patients received a reduced-intensity conditioning regimen with fludarabine, melphalan and thymoglobulin (dose dependent on the degree of HLA matching), in association with cyclosporine and mycophenolate mofetil for GVHD prophylaxis. A total of 21 patients (58%) developed grades II-IV aGVHD in the first six months post-transplant. Whole blood and serum samples were collected from HSCT recipients 1, 2, 3 and 6 months post-HSCT. To assess the impact of different immune markers on the occurrence of aGVHD, patients were categorized into 2 groups based on the development or not of grade II-IV aGVHD in the first six months post-HSCT. Our comprehensive 7-colour flow cytometry panel encompasses phenotypic markers for relevant T, B, NK, NKT and dendritic cell subsets, further including intracellular markers for the assessment of proliferation and susceptibility to apoptosis within regulatory (Treg), conventional (Tcon) CD4 T cells and CD8 T cell subsets. Serum concentrations of IL-2, IL-7, IL-10, IL-15, IFN-γ and TNF-α were evaluated with a multiplex cytokine assay. Absolute counts of lymphocytes, total T cells, CD4 Tcon, CD4 Treg and CD8 T cells display similar reconstitution patterns in unrelated HSCT recipients with and without aGVHD. However, we have observed a clear trend for higher levels of γδ T cells, both in terms of frequency and absolute counts, in patients who develop aGVHD, particularly at 1 month post-HSCT (Mann Whitney test, P = 0.0699). Furthermore, we found a significantly higher frequency of proliferating cells, as assessed by Ki67 expression, within the CD4 T cell population in patients who do not develop aGVHD, both at 2 and 3 months (Mann Whitney test, P = 0.0400 and P = 0.0177, respectively) post-HSCT. When dissected into proliferating CD4 Tcon and Treg, the frequency of both Ki67+ Tcon and Ki67+ Treg within the CD4 T cell population is higher in patients who do not develop aGVHD in all time-points analysed, reaching statistical significance at months 2 and 3 for Tcon (Mann Whitney test: P = 0.0280 and P = 0.0051, respectively) and at month 6 for Treg (Mann Whitney test, P = 0.0043). We are also quantifying the serum levels of relevant pro-inflammatory, suppressive and homeostatic cytokines in the two patient groups. We have observed that IL-7 serum levels (pg/ml) in the first 3 months post-transplant are significantly lower in patients who develop aGVHD compared to those who do not (Mann Whitney test: Month 1 P = 0.0051; Month 2 P = 0.0177; Month 3 P = 0.0303), whereas IL-10 levels are significantly higher in aGVHD patients (Mann Whitney test: Month 1 P = 0.0442; Month 2 P = 0.0051; Month 3 P = 0.0480). None of the other cytokines analyzed has revealed significant differences between the two groups of patients, although there seems to be a trend for higher IL-15 serum levels in patients who develop aGVHD, particularly in the first two months post-HSCT. These results indicate that in our patient population sensitive methods for the detection of IL-7 and IL-10 serum levels can identify patients at high risk for grades II-IV aGVHD early post-transplant. The accrual of more patients will clarify whether the levels of γδ T cells post-transplant, as well as the frequency of proliferating CD4 Tcon and Treg populations, can also be used as an early indicator for the development of aGVHD. Disclosures No relevant conflicts of interest to declare.

Introduction MM is characterized by the presence of a monoclonal immunoglobulin in the serum and/or urine, produced by malignant plasma cells. MM has a highly uncertain evolution and prognostic depending on age, stage and cytogenetic abnormalities. FLC is a readily available laboratory test and has independent prognostic significance in all the plasma cell disease entities helping to clarify International Staging System (ISS). Its value in the era of new therapeutic drugs is to be proved. Purpose To determine the prognostic value of baseline serum FLCr in patients with newly diagnosed MM, consecutively treated with thalidomide or bortezomib based regimens. Methods This is a retrospective study, from July 2004 to December 2012. We studied the relationship of baseline FLCr with outcome and its contribution to clarify the prognostic value of ISS. Serum FLC levels were measured in frozen sera at initial diagnosis, using a latex-enhanced immunoassay (The Binding Site, Birmingham, UK) on a Beckman Coulter nephelometric analyzer. FLCr was calculated as k/λ (reference range: 0,26-1,65). Abnormal FLCr was defined of < 0,03 or > 32,00 and standard FLCr of 0,03-32. Patients have been staged by ISS and Mayo Clinic stratification. All patients received at least two cycles of a thalidomide based regimen, 91 patients (38,9%): thalidomide, doxorubicin, dexamethasone (TAD), 37 patients (15,8%) and melphalan/cyclophosphamide, thalidomide, prednisone (M/CTP), 54 patients (23,0%) or bortezomib based regimens, 143 patients (61,1%): bortezomib, doxorubicin and dexamethasone (PAD), 53 patients (22,6%), bortezomib, cyclophosphamide and dexamethasone (Cy-Bor-D), 7 patients (2,9%) and melphalan/cyclophosphamide, bortezomib, prednisone (M/CVP), 83 patients (35,6%). Median follow-up from diagnosis was 25 months. Follow-up was through the review of each patient’s complete medical records at our center. Statistical analysis was performed with SPSS 20 ®. Results We reviewed 234 patients consecutively treated. Male gender was 46,6% and median age was 67 years (22-84y). The Ig type was IgG in 59,4%, IgA in 22,2%, IgD in 1,7%, light chains MM in 15,4% and non-secretory MM in 1,3%. The median Hb level was 10,7g/dL (4,9-16,7g/dL) and serum albumin was 27,4 g/L (9,0-50-0g/L). Twenty five percent of patients had a creatinine clearance 32,00) and 49,1% had standard FLCr 0,03-32,00. FISH was performed in 74,4% patients: 26,0% presented high-risk features. Overall survival (OS) and progression free survival (PFS) were significantly different according the patients presented 0, 1, 2 or 3 Mayo Clinic risk factors, with median survival times of 83, not reached (NR), 39 and 36 months, for OS, p=0,000, and 41, 41, 17 and 17 months for PFS, p=0,000. Patients with abnormal FLCr had similar outcome of standard FLCr. Patients in ISS II had significant different OS according the FLCr: 77 and 39 months for standard and abnormal FLCr, respectively, P=0,015. There was no such correlation on stages I and III. Conclusions Baseline serum FLCr associated to others risk-factors like serum β2-microglobuline and albumin is a powerful prognostic factor for survival in newly diagnosed MM treated with thalidomide or bortezomib-based therapies. FLCr helps to clarify the outcome of ISS stage II patients identifying a subgroup with clearly inferior outcome. Disclosures: Esteves: Yassen: Consultancy; Celgene: Consultancy. Raposo:Roche; lymphoproliferative diseases: Consultancy. Guerra:Novartis, Bristol-Myers Squibb and Amgen. Advisory bord meetings. Areas involved: Myeloproliferative neoplasms and acute leukaemia: Consultancy.

Introduction MM is a B cell malignancy characterized by the presence of a monoclonal immunoglobulin (Ig) in the serum and/or urine produced by clonal plasma cells. MM has a variable outcome depending on age, stage and cytogenetic abnormalities. FLCr is a readily available laboratory test and has independent prognostic significance in all the plasma cell disease entities, helping to clarify International Staging System (ISS), namely the heterogeneous stage II. In the era of new therapeutic agents trending to personalized therapy, its prognostic value needs to be proved. Purpose To evaluate the prognostic value of baseline serum FLCr on outcome of patients with newly diagnosed MM eligible for HDM treated with thalidomide or bortezomib based regimens. Methods This is a retrospective study, from January 2005 to December 2012. We analyzed the relationship of baseline FLCr with outcome and its contribution to clarify the prognostic value of ISS. Serum FLC levels were measured in frozen sera drew at diagnosis before treatment, using a latex-enhanced immunoassay (The Binding Site, Birmingham, UK) on a Beckman Coulter nephelometric analyzer. FLCr was calculated as k/λ (reference range: 0,26-1,65). Abnormal FLCr was defined of 32,00 and standard FLCr of 0,03-32. Patients have been staged by ISS and Mayo Clinic stratification. All patients received at least two cycles of a bortezomib based regimen (64,7%), with doxorubicin and dexamethasone (PAD) in 55 patients (53,9%), with cyclophosphamide and dexamethasone (Cy-Bor-D) in 11 patients (10,8%) or thalidomide, doxorubicin and dexamethasone (TAD) in 36 patients (35,3%). Median follow-up from diagnosis was 30 months. Follow-up was through the review of each patient’s complete medical records at our center. Statistical analysis was performed with SPSS 20 ®. Results We reviewed 102 patients treated with thalidomide or bortezomib based-regimens eligible for HDM. Fifty one percent of patients were male and median age was 52 years (22-70y). The Ig type was IgG in 62,7%, IgA in 14,7%, IgD in 3,9%, light chains MM in 17,7% and non-secretory MM in 1,0% of patients. The median Hb level was 9,8 g/dL (5,3-16,7mg/L) and median serum albumin was 31,5 g/L (22,3-48,3g/L). Creatinine clearance 32,00). Cytogenetic FISH analysis was performed in 86,3% of patients: 32,4% presented high-risk features. We observed a significant difference on overall survival (OS) and progression free survival (PFS) according to Mayo Clinic risk-factors: 0, not reached (NR), 1, NR, 2, 49 months and 3, 71 months for OS (P=0,000) and 0, 68 months, 1, 55 months, 2, 16 months and 3, 24 months for PFS (P=0,000). According the FLCr there was a significant advantage on OS for standard FLCr: NR vs 71 months (P=0,035). A trend to a better OS (P=0,061) was observed in stage II patients with standard FLCr but no significant difference at any stage. Conclusions Baseline serum FLCr associated to others risk-factors like serum β2-microglobuline and albumin is a powerful prognostic factor for survival in newly diagnosed MM eligible for HDM treated with thalidomide or bortezomib-based therapies. We fail to fully demonstrate FLCr prognostic value on ISS stage II patients, only a trend for better outcome with standard FLCr. Disclosures: Esteves: Janssen-Cylag and Celgene. Consultancy on the area of multiple myeloma and acute myeloid leukaemia: Consultancy. Raposo:Roche; lymphoproliferative diseases: Consultancy. Guerra:Novartis, Bristol-Myers Squibb and Amgen. Advisory bord meetings. Areas involved: Myeloproliferative neoplasms and acute leukaemia: Consultancy.