EOMES and IL-10 regulate antitumor activity of T regulatory type 1 CD4 + T cells in chronic lymphocytic leukemia

The transcription factor eomesodermin (EOMES) promotes interleukin (IL)-10 expression in CD4(+) T cells, which has been linked to immunosuppressive and cytotoxic activities. We detected cytotoxic, programmed cell death protein-1 (PD-1) and EOMES co-expressing CD4(+) T cells in lymph nodes (LNs) of patients with chronic lymphocytic leukemia (CLL) or diffuse large B-cell lymphoma. Transcriptome and flow cytometry analyses revealed that EOMES does not only drive IL-10 expression, but rather controls a unique transcriptional signature in CD4(+) T cells, that is enriched in genes typical for T regulatory type 1 (T(R)1) cells. The T(R)1 cell identity of these CD4(+) T cells was supported by their expression of interferon gamma and IL-10, as well as inhibitory receptors including PD-1. T(R)1 cells with cytotoxic capacity accumulate also in Eµ-TCL1 mice that develop CLL-like disease. Whereas wild-type CD4(+) T cells control TCL1 leukemia development after adoptive transfer in leukopenic Rag2(−/)(−) mice, EOMES-deficient CD4(+) T cells failed to do so. We further show that T(R)1 cell-mediated control of TCL1 leukemia requires IL-10 receptor (IL-10R) signaling, as Il10rb-deficient CD4(+) T cells showed impaired antileukemia activity. Altogether, our data demonstrate that EOMES is indispensable for the development of IL-10-expressing, cytotoxic T(R)1 cells, which accumulate in LNs of CLL patients and control TCL1 leukemia in mice in an IL-10R-dependent manner

Fantastic voyage: the journey of intestinal microbiota-derived microvesicles through the body

As part of their life cycle, Gram-negative bacteria produce and release microvesicles (outer membrane vesicles, OMVs) consisting of spherical protrusions of the outer membrane that encapsulate periplasmic contents. OMVs produced by commensal bacteria in the gastrointestinal (GI) tract of animals are dispersed within the gut lumen with their cargo and enzymes being distributed across and throughout the GI tract. Their ultimate destination and fate is unclear although they can interact with and cross the intestinal epithelium using different entry pathways and access underlying immune cells in the lamina propria. OMVs have also been found in the bloodstream from which they can access various tissues and possibly the brain. The nanosize and non-replicative status of OMVs together with their resistance to enzyme degradation and low pH, alongside their ability to interact with the host, make them ideal candidates for delivering biologics to mucosal sites, such as the GI and the respiratory tract. In this mini-review, we discuss the fate of OMVs produced in the GI tract of animals with a focus on vesicles released by Bacteroides species and the use of OMVs as vaccine delivery vehicles and other potential applications

Regulación de los linfocitos B y T yd+ sobre el repertorio T aB+ específico de superantígenos endógenos, presentados en condiciones de baja afinidad

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 21-03-200

Bases para la medida de un aprendizaje matemático significativo

Construir la didáctica de la matemática como ciencia fundamental autónoma, que debe crear simultáneamente sus objetos de estudio y sus métodos, los cuales no deben ser los mismos de las ciencias experimentales. Muestra aleatoria compuesta por 30 estudiantes de primero de la especialidad de Ciencias. Propone un modelo de aprendizaje-enseñanza de las matemáticas que desarrolla y potencia más el pensamiento matemático. Aplica pruebas para medir la aptitud de los alumnos para resolver problemas y constatar la comprensión. Se basa en el análisis experimental. Utiliza la Hoja de Cálculo Lotus 1-2-3, el Turbo Basic para hacer programas estadísticos de cálculos y representaciones gráficas, el procesador de textos WP5.1, y los programas Story, Picture Maker, Picture Taker, Story Editor, Story Teller y Text Maker.. Se detecta un desajuste del acto didáctico en el aprendizaje-enseñanza de las Matemáticas. Se confirma que el razonamiento matemático es un proceso dinámico y secuenciado en el espacio y en el tiempo. Así, la competencia de un estudiante en el conocimiento de una materia tan específica como la matemática depende del camino recorrido en su trayectoria educativa.MadridES

Foxp3⁺ cells upregulate Nur77 after TCR activation in an antigen-specific manner.

<p>(A) Time course of Nur77 (black bars) and CD69 expression (white bars) on Foxp3⁺ T-cells among sorted CD25⁺ CD4⁺ splenocytes stimulated with plate-bound αCD3 and 1U/ml IL-2. Data are representative of 3 independent experiments with n ≥ 3. (B) Nur77 expression on Foxp3^- (left) and Foxp3⁺ (right) among enriched CD4⁺ T-cells from either wild-type (white bars) or OT-II (black bars) mice, co-cultured overnight with unstimulated, OVA pulsed or αCD3 coated BMDCs. Data are representative of two independent experiments, with each n = 3 per group. unstim, control without BMDC; /, unpulsed BMDC; OVA, OVA-pulsed BMDC, anti-CD3, anti-CD3-coated BMDC. Data show mean + SD, *p ≤ 0.05, *** p ≤ 0.001; ns, non significant. Data are representative of four independent experiments each with n = 3 per group.</p

Foxp3⁺ cells preferentially upregulate CD69 in response to soluble factors.

<p>(A) CD69 expression on Foxp3^- (left) and Foxp3⁺ (right) enriched CD4⁺ T cells after overnight incubation with or without BMDC-conditioned culture medium. /, control medium; OVA, medium supplemented with OVA; BMDC SN, supernatant from BMDCs cultured overnight with fresh medium; BMDC OVA SN, supernatant from BMDCs cultured overnight with medium supplemented with OVA. Data are representative of at least two independent experiments. (B) CD69 expression on Foxp3^- (white bars) and Foxp3⁺ (black bars) among enriched CD4⁺ T cells cultured overnight without additional cytokines (/) or with IL-1β, IFN-α or TNF-α. Data are representative of at least two independent experiments. (C) CD69 expression on Foxp3⁺ among enriched CD4⁺ T cells cultured overnight without additional cytokines (/) or with IL-33, IL-4, IL-12, IL-27, IL-6, IFN-γ, or GM-CSF. (D) Representative FACS plot and response to stimulation of sorted CD4⁺ Foxp3^RFP+ cells. Right: representative FACS plot showing the purity of Foxp3^RFP+ cells after sort. Cells are gated on forward and side scatter and doublets and dead cells are excluded as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137393#pone.0137393.g001" target="_blank">Fig 1A</a>. Plot shows CD4 versus intracellular Foxp3 staining. Left, CD69 expression on Foxp3⁺ among enriched CD4⁺ T cells (white bars) or Foxp3^RFP+ sorted (black bars) CD4⁺ T cells, cultured overnight without additional cytokines (/) or with IFN-α or TNF-α (left). (E) CD69 expression on Foxp3⁺ among enriched CD4⁺ splenocytes from TNFR1 KO, Myd88 KO and ctrl C57BL/6 or from IFNAR1 KO and control 129 mice, cultured overnight with control medium (/) or with supernatant from OVA stimulated BMDCs (BMDC OVA SN). (F) CD69 expression on Foxp3⁺ among enriched CD4⁺ T cells of IFNAR1 <i>KO</i> and control mice cultured overnight with control medium (unstim) or with supernatant from OVA stimulated BMDCs (BMDC OVA SN) with or without addition of different concentrations of blocking anti-TNF-α antibody. Data are representative from two independent experiments. Data show mean + SD, *** p ≤ 0.001 compared to unstimulated control, unless comparison indicated by line below the stars, n ≥ 3 per group.</p

Foxp3⁺ cells upregulate CD69 after TCR activation in an antigen-non-specific manner.

<p>(A) Gating strategy on CD4⁺ enriched T cells after overnight culture without stimulation. Cells are gated in the lymphocyte gate based on forward versus side scatter, then doublets are excluded based on forward scatter height versus amplitude and live cells are gated as negative for the dead cell marker. Finally Foxp3⁺ and Foxp3^- CD4⁺ T cells are gated using CD4 and Foxp3 expression. This gating strategy was applied throughout the analyses unless otherwise stated. (B) CD69 expression on Foxp3^- (top row) and Foxp3⁺ (bottom row) CD4⁺ cells before and after overnight stimulation with or without anti-CD3 stimulation. Staining is representative of at least three independent experiments. (C) CD69 expression on Foxp3^- (left) and Foxp3⁺ (right) enriched CD4⁺ T cells from either wild-type (white bars) or OT-II (black bars) donor mice, after overnight culture with BMDCs. Unstim, control without BMDC; /, unpulsed BMDC; OVA, OVA-pulsed BMDC, anti-CD3, anti-CD3-coated BMDC. Data are representative of four independent experiments with each 3 replicates per group. (D) Normalized MHCII KO BMDC-induced CD69 expression compared to control BMDC-induced CD69. The graphs show the frequency of CD69⁺ cells among CD4⁺ Foxp3^- (left) and Foxp3⁺ (right) T cells after stimulation with MHCII KO BMDC, normalized to CD4⁺ Foxp3^- and Foxp3⁺ T cells stimulated with wild type BMDC for each given condition. 100% represents the same CD69 expression by cells stimulated with control or MHCII KO BMDC. Data show mean + SD, *p ≤ 0.05; **p ≤ 0.01; *** p ≤ 0.001, ns, non significant. Data are representative of three independent experiments with each n = 3 per group.</p

Nur77 intensity is not modulated by cytokines.

<p>(A) Nur77 expression on Foxp3^- (white bars) and Foxp3⁺ (black bars) CD4⁺ T cells from the culture in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137393#pone.0137393.g002" target="_blank">Fig 2B</a>. Data are representative of at least two independent experiments. (B) Representative plots of Nur77 versus CD69 expression on Foxp3⁺ among enriched CD4⁺ T cells, stimulated for 4 hours (top row) or 16 hours (bottom row) without additional cytokines (/), with IFN-α, TNF-α, or plate-bound αCD3. Data are representative of two independent experiments. (C) Nur77 expression on Foxp3⁺ cells among sorted CD25⁺ CD4⁺ splenocytes stimulated with plate-bound αCD3 with low (1U/ml) or high (1000U/ml) IL-2 for 6 hours. Data are representative of four independent experiments. Data show mean + SD, ns, non significant.</p

El desarrollo psicológico del niño de de 3 a 6 años

Resumen tomado de la web del Departamento de EducaciónPresenta unas indicaciones sobre el desarrollo psicomotor, el desarrollo de la inteligencia,el desarrollo del lenguaje y el desarrollo afectivo social. También ofrece una serie de orientaciones y consejos a los padres sobre el desarrollo de cada uno de estos aspectos. Está ilustrado con unos atractivos dibujos.NavarraBiblioteca General de Navarra; Calle Plaza San Francisco, s. n.; 31001 Pamplona;ES