Poli (ADP-Ribosa) Polimerasa-1: una proteína nuclear implicada en procesos inflamatorios, muerte celular y cáncer

Numerosos estudios en modelos experimentales han puesto de manifiesto que el bloqueo genético o lainhibición farmacológica de poli-ADP-ribosa-polimerasa-1, proteína nuclear implicada en fenómenos deseñalización celular a través de modificaciones postraduccionales mediante poli-ADP-ribosilación, confiereprotección frente a procesos citolíticos derivados que tienen lugar durante el desarrollo de la respuestainflamatoria. Un denominador común en todos los procesos inflamatorios es la secreción de diversosmediadores proinflamatorios y la formación de radicales libres que van a desencadenar la activación de poli-ADP-ribosa-polimerasa-1 y simultáneamente se potencia la activación de diversos factores de transcripcióncomo NF-kB y AP-1, dando lugar a la expresión de genes dependientes de éstos. Es bien conocido que lainflamación en el cáncer, como proceso de estrés oxidativo continuo, actúa como un fuerte promotor tumoralfavoreciendo el desarrollo del tumor. Esta revisión pretende dar una visión general sobre el conocimientoactual de esta proteína tanto a nivel celular como en procesos patológicos tan importantes como el cáncer. PALABRAS CLAVE: PARP-1, Inflamación, Cáncer, Factores de Transcripción

Pluripotent stem cell regulation in Spain and the Spanish National Stem Cell Bank.

The Spanish National Stem Cell Bank (Banco Nacional de Líneas Celulares, BNLC) was established in 2006 thanks to a change in the legislative framework in Spain. The Law 14/2006 updated the previous Assisted Reproduction Techniques Law (Law 45/2003) allowing the use of the surplus frozen embryos following IVF for research. The BNLC has a network structure with 3 nodes: the Regenerative Medicine Program (IDIBELL), the Principe Felipe Research Center (CIPF) in Valencia and the Andalusian Public Health System Biobank (SSPA Biobank) in Granada. The aim of the BNLC is to guarantee throughout the national territory the availability of human stem cell lines for biomedical research. At present time, there are 40 human embryonic stem cell lines (hESC) and 171 human induced pluripotent stem cell lines (hiPSC) registered in the BNLC. These lines are fully characterized and available in the context of research projects approved by the Technical Committee of the BNLC.The National Spanish Stem Cell Bank is supported by the Plataforma de Proteomica, Genotipado y Líneas Celulares (PT1770019/0015) (PRB3), Instituto de Salud Carlos III.S

Stabilization of Human Whole Blood Samples for Multicenter and Retrospective Immunophenotyping Studies

Whole blood is often collected for large-scale immune monitoring studies to track changes in cell frequencies and responses using flow (FC) or mass cytometry (MC). In order to preserve sample composition and phenotype, blood samples should be analyzed within 24 h after bleeding, restricting the recruitment, analysis protocols, as well as biobanking. Herein, we have evaluated two whole blood preservation protocols that allow rapid sample processing and long-term stability. Two fixation buffers were used, Phosphoflow Fix and Lyse (BD) and Proteomic Stabilizer (PROT) to fix and freeze whole blood samples for up to 6 months. After analysis by an 8-plex panel by FC and a 26-plex panel by MC, manual gating of circulating leukocyte populations and cytokines was performed. Additionally, we tested the stability of a single sample over a 13-months period using 45 consecutive aliquots and a 34-plex panel by MC. We observed high correlation and low bias toward any cell population when comparing fresh and 6 months frozen blood with FC and MC. This correlation was confirmed by hierarchical clustering. Low coefficients of variation (CV) across studied time points indicate good sample preservation for up to 6 months. Cytokine detection stability was confirmed by low CVs, with some differences between fresh and fixed conditions. Thirteen months regular follow-up of PROT samples showed remarkable sample stability. Whole blood can be preserved for phenotyping and cytokine-response studies provided the careful selection of a compatible antibody panel. However, possible changes in cell morphology, differences in antibody affinity, and changes in cytokine-positive cell frequencies when compared to fresh blood should be considered. Our setting constitutes a valuable tool for multicentric and retrospective studies. (c) 2020 International Society for Advancement of Cytometr

IL8 and EDEM3 gene expression ratio indicates peripheral blood mononuclear cell (PBMC) quality

Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs)

IL8 and IL16 levels indicate serum and plasma quality

Longer pre-centrifugation times alter the quality of serum and plasma samples. Markers for such delays in sample processing and hence for the sample quality, have been identified. Twenty cytokines in serum, EDTA plasma and citrate plasma samples were screened for changes in concentration induced by extended blood pre-centrifugation delays at room temperature. The two cytokines that showed the largest changes were further validated for their "diagnostic performance" in identifying serum or plasma samples with extended pre-centrifugation times. In this study, using R&D Systems ELISA kits, EDTA plasma samples and serum samples with a pre-centrifugation delay longer than 24 h had an IL16 concentration higher than 313 pg/mL, and an IL8 concentration higher than 125 pg/mL, respectively. EDTA plasma samples with a pre-centrifugation delay longer than 48 h had an IL16 concentration higher than 897 pg/mL, citrate plasma samples had an IL8 concentration higher than 21.5 pg/mL and serum samples had an IL8 concentration higher than 528 pg/mL. These robust and accurate tools, based on simple and commercially available ELISA assays can greatly facilitate qualification of serum and plasma legacy collections with undocumented pre-analytics

Integration of cytokine profiles with distinct B-cell functions reveals a specific micro-environmental framework in autoimmune diseases

International audienc

Integration of cytokine profiles with distinct B-cell functions reveals a specific micro-environmental framework in autoimmune diseases

International audienc

A cytokine network profile delineates a common Th1/Be1 pro‐inflammatory group of patients in four systemic autoimmune diseases

International audienc