Implicación de la pseudoquinasa IRAK-M en los procesos de tolerancia a endotoxina y a tumores

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina. Departamento de Bioquímica. Fecha de lectura: 13 de Diciembre de 200

Deficient neutralizing antibody response and specific lack of RBD-responsive B cells in elderly long-term convalescent patients from severe COVID-19

Resumen del póster presentado a las III Jornadas Científicas PTI+ Salud Global, celebradas en el Centro de Ciencias Humanas y Sociales (CCHS), CSIC (Madrid) del 20 al 22 de noviembre de 2023.[Background] Severe COVID-19 is defined by admission to intensive care units with respiratory support. This condition increases with age. To investigate in the possible mechanisms by which severe COVID-19 occurs, we compared the response of specific antibodies against the viral antigens RBD, Spike (S), nuclear (N), and membrane (Mpro) and the neutralizing titers in plasma of elderly patients convalescent from severe COVID-19 with convalescent patients from mild disease.[Methods] Plasma was collected from cohort 1: 20 healthy donors (vaccinated and unvaccinated), cohort 2: 40 elderly long-term convalescent patients from severe COVID-19 (mean: 73 years old, 60/40 ratio men/women, and 10 months after infection), and cohort 3: 60 convalescent from mild COVID-19 patients, who were vaccinated (mean: 42 years old, 30/70 ration men/women, 8 months after mild infection, and 5.7 months after last vaccination shoot). To compare reactivity against native S protein, we used a sensitive method to measure specific IgG1 and IgA in plasma by flow cytometry. We assessed the neutralization titers in plasma by infection assays in HEK-293T cells or Vero cells expression ACE-2, using S- and GFP-expressing pseudotyped viruses, and quantified IgG, IgA, and IgM antibodies against RBD, S, N, and mPro antigens using the Multiplex Serological SARS-CoV-2 assay (Immunostep). To evaluate the presence of specific T and B cells specific for S and RBD antigens, we used kits from Miltenyi Biotech and analyzed the expression of activation antigens after cell stimulation with these antigens by flow cytometry.[Results] Significant levels of IgG and IgA against S protein were found in the plasma of convalescent patients from cohorts 2 and 3, with no differences in total anti-S antibody response between the two cohorts. Interestingly, anti-RBD IgG levels were found to be extremely low in plasma samples from cohort 2, but not in plasma samples from cohort 3. In accordance with these results, the neutralizing titers (IC50) found were very low in the plasma samples from cohort 2, compared to those from cohort 3. Analysis of the presence of RBD- and S-specific B cells present in peripheral blood mononuclear cells (PBMCs) of these cohorts revealed significantly lower levels of RBD-specific B cells, but of not S-specific B lymphocytes, in the PBMCs from cohort 2 cells but not in those from cohort 3. Furthermore, lower B cell activation, as demonstrated by CD25 expression, was observed in PBMCs from cohort 2 compared to those from cohort 3 after their stimulation with RBD, but not with S, N or Mpro proteins. These results together indicate the specific lack of RBD-specific B cells in cohort 2 patients.[Conclusions] The low neutralizing capacity observed in the plasma of elderly long-term convalescent patients, who recovered from severe COVID-19 (cohort 2) correlates with low specific levels of anti-RBD antibodies and reduced levels of RBD-responsive B cells. These results could help explain the severity of COVID-19 in patients from cohort 2 compared to those from cohort 3, who had a mild disease. Future experiments will evaluate the presence of neutralizing antibodies in cohort 2 patients after vaccination.Peer reviewe

DRI Triton SS-OCT applied to detect choroidal nodules in paediatric patients affected by NF1

Purpose: To examine whether image processing of non-mydriatic DRI Triton SS-OCT (Topcon Corporation, Tokyo, Japan) using the red free filter could assess the presence of choroidal nodules and thus include their detection as a diagnostic criterion in neurofibromatosis type 1 (NF1). Material and methods: We included 417 eyes from 210 patients, 377 - from 190 patients diagnosed with NF1 according to the criteria established by the National Institutes of Health Consensus Development Conference (NIH) and 40 from 20 healthy patients as a control group. The mean age was 9.4 years (range 2 years–18 years). All patients had their visual acuity measured by a test according to age, were examined for the presence of lisch nodules and an Optical Coherence Tomography (OCT) of the macular area was performed. All the OCT images were analysed to check if visible nodules could be identified. Results: Ages 14 (95% CI=(9.7,18.3)) and 12 years (95% CI=(9.1,14)) are the cut-off points that best separate those with choroidal nodules with Triton OCT and lisch with slit lamp, respectively, from those without. lisch nodules were detected in 50% of cases of NF1 patients. The presence of choroidal nodules did not present a statistically significant correlation with the occurrence of optic pathway glioma (p = 0.96) nor with the patient's visual worsening (p = 0.072). A statistically significant correlation was observed between the presence of choroidal nodules and the presence of lisch nodules (p < 0.05). Conclusion: The Topcon Triton OCT red free tool would not be a good tool to detect choroidal nodules in patients with NF1 because of its low sensitivity. If the presence of choroidal nodules were to be included in the diagnostic criteria for NF1, it would be convenient to use a device with red and infrared radiations

Fused Cells between Human-Adipose-Derived Mesenchymal Stem Cells and Monocytes Keep Stemness Properties and Acquire High Mobility

Human-adipose-derived mesenchymal stem cells (hADMSCs) are multipotent stem cells which have become of great interest in stem-cell therapy due to their less invasive isolation. However, they have limited migration and short lifespans. Therefore, understanding the mechanisms by which these cells could migrate is of critical importance for regenerative medicine. Methods: Looking for novel alternatives, herein, hADMSCs were isolated from adipose tissue and co-cultured with human monocytes ex vivo. Results: A new fused hybrid entity, a foam hybrid cell (FHC), which was CD90+CD14+, resulted from this co-culture and was observed to have enhanced motility, proliferation, immunomodulation properties, and maintained stemness features. Conclusions: Our study demonstrates the generation of a new hybrid cellular population that could provide migration advantages to MSCs, while at the same time maintaining stemness properties

DNGR-1 limits Flt3L-mediated antitumor immunity by restraining tumor-infiltrating type I conventional dendritic cells.

BackgroundConventional type 1 dendritic cells (cDC1s) are central to antitumor immunity and their presence in the tumor microenvironment associates with improved outcomes in patients with cancer. DNGR-1 (CLEC9A) is a dead cell-sensing receptor highly restricted to cDC1s. DNGR-1 has been involved in both cross-presentation of dead cell-associated antigens and processes of disease tolerance, but its role in antitumor immunity has not been clarified yet.MethodsB16 and MC38 tumor cell lines were inoculated subcutaneously into wild-type (WT) and DNGR-1-deficient mice. To overexpress Flt3L systemically, we performed gene therapy through the hydrodynamic injection of an Flt3L-encoding plasmid. To characterize the immune response, we performed flow cytometry and RNA-Seq of tumor-infiltrating cDC1s.ResultsHere, we found that cross-presentation of tumor antigens in the steady state was DNGR-1-independent. However, on Flt3L systemic overexpression, tumor growth was delayed in DNGR-1-deficient mice compared with WT mice. Of note, this protection was recapitulated by anti-DNGR-1-blocking antibodies in mice following Flt3L gene therapy. This improved antitumor immunity was associated with Batf3-dependent enhanced accumulation of CD8+ T cells and cDC1s within tumors. Mechanistically, the deficiency in DNGR-1 boosted an Flt3L-induced specific inflammatory gene signature in cDC1s, including Ccl5 expression. Indeed, the increased infiltration of cDC1s within tumors and their protective effect rely on CCL5/CCR5 chemoattraction. Moreover, FLT3LG and CCL5 or CCR5 gene expression signatures correlate with an enhanced cDC1 signature and a favorable overall survival in patients with cancer. Notably, cyclophosphamide elevated serum Flt3L levels and, in combination with the absence of DNGR-1, synergized against tumor growth.ConclusionDNGR-1 limits the accumulation of tumor-infiltrating cDC1s promoted by Flt3L. Thus, DNGR-1 blockade may improve antitumor immunity in tumor therapy settings associated to high Flt3L expression

A-BASE-DE-PROS: una implementación práctica de los Objetivos de Desarrollo Sostenible en la Universidad Politécnica de Madrid

A lo largo de los últimos años hemos visto cómo los Objetivos de Desarrollo Sostenible (ODS) han ido permeando las distintas capas de la sociedad, estableciendo nuevas prioridades tanto en las políticas públicas como en las empresariales. La educación no ha quedado ajena a este cambio, sino que también se está alineando con las metas anteriores. En este capítulo se describen las principales actividades realizadas en el marco del proyecto APS22.2003 “Aprendizaje BAsado en SErvicio DE ODS relacionados con una PROducción y consumo responsableS (A-BASE-DE-PROS)”, en el que el ODS 12 se toma como eje central para concienciar a estudiantes universitarios y de secundaria de la importancia de la Agenda 2030. En general, hemos comprobado que el proyecto ha permitido dar a conocer los ODS y que la mayoría de los estudiantes, tanto universitarios como de secundaria, han valorado la experiencia como positiva.

Tumor stem cells fuse with monocytes to form highly invasive tumor-hybrid cells

The ‘cancer cell fusion’ theory is controversial due to the lack of methods available to identify hybrid cells and to follow the phenomenon in patients. However, it seems to be one of the best explanations for both the origin and metastasis of primary tumors. Herein, we co-cultured lung cancer stem cells with human monocytes and analyzed the dynamics and properties of tumor-hybrid cells (THC), as well as the molecular mechanisms beneath this fusion process by several techniques: electron-microscopy, karyotyping, CRISPR-Cas9, RNA-seq, immunostaining, signaling blockage, among others. Moreover, mice models were assessed for in vivo characterization of hybrids colonization and invasiveness. Then, the presence of THCs in bloodstream and samples from primary and metastatic lesions were detected by FACS and immunofluorescence protocols, and their correlations with TNM stages established. Our data indicate that the generation of THCs depends on the expression of CD36 on tumor stem cells and the oxidative state and polarization of monocytes, the latter being strongly influenced by microenvironmental fluctuations. Highly oxidized M2-like monocytes show the strongest affinity to fuse with tumor stem cells. THCs are able to proliferate, colonize and invade organs. THC-specific cell surface signature CD36+CD14+PANK+ allows identifying them in matched primary tumor tissues and metastases as well as in bloodstream from patients with lung cancer, thus functioning as a biomarker. THCs levels in circulation correlate with TNM classification. Our results suggest that THCs are involved in both origin and spread of metastatic cells. Furthermore, they might set the bases for future therapies to avoid or eradicate lung cancer metastasis.Research in the laboratory of EL-C was supported by “Fundación para la Investigación Biomédica La Paz Hospital” ((FIBHULP) and “Fundación Familia Alonso”.Peer reviewe

Experiencias de Aprendizaje-Servicio en la UPM: 2021 y 2022

La Oficina de Aprendizaje-Servicio (ApS) de la UPM, constituida en sesión del Consejo de Gobierno de diciembre de 2019 tiene, como misión fundamental, promover en el ámbito de las enseñanzas de esta universidad la metodología ApS. Con esta finalidad se vienen realizando convocatorias de proyectos de impacto social alineados con los ODS como un mecanismo más para la contribución a la Agenda 2030, y se colabora intensamente con las diversas oficinas constituidas con el mismo objetivo en otras universidades. Nuestra oficina pretende impulsar progresivamente la colaboración con entidades ajenas a la UPM, y atender demandas y necesidades sociales en las que nuestros estudiantes y profesores brinden sus conocimientos para la construcción de una mejor y más justa sociedad. Con este propósito, se han puesto en marcha numerosas iniciativas y colaboraciones con Ayuntamientos, Asociaciones, ONG, Fundaciones y centros de enseñanza, con el fin común de plantear mejoras y trabajar con entornos desfavorecidos, y colectivos vulnerables de nuestro entorno. Cabe destacar la muy positiva acogida que, progresivamente se está logrando, en lo relativo a la diseminación de estas iniciativas en el ámbito de la UPM, viéndose incrementada la participación e interés de nuestros docentes y estudiantes en los llamamientos que se realizan desde la oficina. Desde la constitución de la oficina, son ya más de 100 actividades desarrolladas con la participación de más de 500 profesores. Uno de los compromisos de la Oficina ApS de la UPM es dar visibilidad por su carácter meritorio a las experiencias realizadas por el profesorado y los estudiantes de nuestra universidad y, es por ello, que nos complace la presentación de esta primera edición del ebook, en el que se recogen algunas de las experiencias realizadas en nuestra universidad y que confiamos ampliar periódicamente con futuras ediciones. Nuestro más sincero agradecimiento a todos los profesores que habéis hecho posible esta primera publicación con vuestras contribuciones