ABSTRACT
Naked mole‐rats express many unusual traits for such a small rodent. Their morphology, social behaviour, physiology, and ageing have been well studied over the past half‐century. Many early ...findings and speculations about this subterranean species persist in the literature, although some have been repeatedly questioned or refuted. While the popularity of this species as a natural‐history curiosity, and oversimplified story‐telling in science journalism, might have fuelled the perpetuation of such misconceptions, an accurate understanding of their biology is especially important for this new biomedical model organism. We review 28 of these persistent myths about naked mole‐rat sensory abilities, ecophysiology, social behaviour, development and ageing, and where possible we explain how these misunderstandings came about.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
CD4+ regulatory T cells (Tregs) are key mediators of immunological tolerance and promising effector cells for immuno-suppressive adoptive cellular therapy to fight autoimmunity and chronic ...inflammation. Their functional stability is critical for their clinical utility and has been correlated to the demethylated state of the TSDR/CNS2 enhancer element in the Treg lineage transcription factor FOXP3. However, proof for a causal contribution of the TSDR de-methylation to FOXP3 stability and Treg induction is so far lacking. We here established a powerful transient-transfection CRISPR-Cas9-based epigenetic editing method for the selective de-methylation of the TSDR within the endogenous chromatin environment of a living cell. The induced de-methylated state was stable over weeks in clonal T cell proliferation cultures even after expression of the editing complex had ceased. Epigenetic editing of the TSDR resulted in FOXP3 expression, even in its physiological isoform distribution, proving a causal role for the de-methylated TSDR in FOXP3 regulation. However, successful FOXP3 induction was not associated with a switch towards a functional Treg phenotype, in contrast to what has been reported from FOXP3 overexpression approaches. Thus, TSDR de-methylation is required, but not sufficient for a stable Treg phenotype induction. Therefore, targeted demethylation of the TSDR may be a critical addition to published
Treg induction protocols which so far lack FOXP3 stability.
Human B lymphocytes are attractive targets for immunotherapies in autoantibody-mediated diseases. Gene editing technologies could provide a powerful tool to determine gene regulatory networks ...regulating B cell differentiation into plasma cells, and identify novel therapeutic targets for prevention and treatment of autoimmune disorders. Here, we describe a new approach that uses CRISPR-Cas9 technology to target genes in primary human B cells in vitro for identifying plasma cell regulators. We found that sgRNA and Cas9 components can be efficiently delivered into primary human B cells through RD114-pseudotyped retroviral vectors. Using this system, we achieved approximately 80% of gene knockout efficiency. We disrupted expression of a triad of transcription factors, IRF4, PRDM1, and XBP1, and showed that human B cell survival and plasma cell differentiation are severely impaired. Specifically, that IRF4, PRDM1, and XBP1 were expressed at different stages during plasma cell differentiation, IRF4, PRDM1, and XBP1-targeted B cells failed to progress to the pre-plasmablast, plasma cell state, and plasma cell survival, respectively. Our method opens a new avenue to study gene functions in primary human B cells and identify novel plasma cell regulators for therapeutic applications.
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Here, we describe a new approach that uses CRISPR-Cas9 technology to target genes in primary human B cells in vitro for identifying plasma cell regulators. We disrupted expression of a triad of transcription factors, IRF4, PRDM1, and XBP1, and showed that human B cell survival and plasma cell differentiation are severely impaired.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Adoptive transfer of regulatory T cells (Treg) is a promising new therapeutic option to treat detrimental inflammatory conditions after transplantation and during autoimmune disease. To reach ...sufficient cell yield for treatment,
ex vivo
isolated autologous or allogenic Tregs need to be expanded extensively
in vitro
during manufacturing of the Treg product. However, repetitive cycles of restimulation and prolonged culture have been shown to impact T cell phenotypes, functionality and fitness. It is therefore critical to scrutinize the molecular changes which occur during T cell product generation, and reexamine current manufacturing practices. We performed genome-wide DNA methylation profiling of cells throughout the manufacturing process of a polyclonal Treg product that has proven safety and hints of therapeutic efficacy in kidney transplant patients. We found progressive DNA methylation changes over the duration of culture, which were donor-independent and reproducible between manufacturing runs. Differentially methylated regions (DMRs) in the final products were significantly enriched at promoters and enhancers of genes implicated in T cell activation. Additionally, significant hypomethylation did also occur in promoters of genes implicated in functional exhaustion in conventional T cells, some of which, however, have been reported to strengthen immunosuppressive effector function in Tregs. At the same time, a set of reported Treg-specific demethylated regions increased methylation levels with culture, indicating a possible destabilization of Treg identity during manufacturing, which was independent of the purity of the starting material. Together, our results indicate that the repetitive TCR-mediated stimulation lead to epigenetic changes that might impact functionality of Treg products in multiple ways, by possibly shifting to an effector Treg phenotype with enhanced functional activity or by risking destabilization of Treg identity and impaired TCR activation. Our analyses also illustrate the value of epigenetic profiling for the evaluation of T cell product manufacturing pipelines, which might open new avenues for the improvement of current adoptive Treg therapies with relevance for conventional effector T cell products.
The project Saadallah Wannous cultural constitutes progressive project at the Arab level. In view of the busy worries of rights issues differentiated, and Considered his experience in the political ...theater of the mature tests, especially those discussed the Palestinian issue and the reality of the Palestinian human rights under occupation .especially the reality of women. Saadalla Wannous in his Play (Alightissab) (Rape) focused on the concept of the Arab Israeli Struggle through his personal contemplation in "Palestine" the essential Question of the Arabs. He called for the reconsideration of this Question in a democratic way far from scare and taboo and the surrounding structural environment. He emphasized on the importance of dealing with this Question within a real and substantial concept of the nature of the Arab-Zionist struggle and understanding the Zionist ideology correctly , different from the previous one. This is what the Author aimed at in this Play that bears new indicative signs the most of important of which is the feminine opposition between the narrations, the Israeli and the Arab, and its impact on the intellectual and social building up. . In The Rape Wannous portrays his feminine Palestinian and Jewish characters realistically, logically and objectively. The Play was an attempt to understand dimensions and ills of the Jewish characters and the stubbornness of the Palestinian with itsdetermination to survive despite theengulfing frustration.
Saadalla Wannous had no embarrassment in expressing the individual tendencies and inclinations which he used to consider bourgeoisie matters. He started to feel with a kind of freedom , which he ...always believed in, which was the substantial condition for the realization of the Stage efficiency, and which he missed in his previous creativities. Previously, he used to impose on himself self-censorship which obliged him to overpass what is personal and individual. That is why we see him in his Play (Tokous Alisharat Wa Altahawolat) (Rites of Signs and Conversions) moves from the general condition to the individual structure within the context of the community. He strictly refers in this Play to the concept of the individual freedom through the psychological structure and subduing the will potential in the human self. Thus, he made his characters express their inner selves far from the social preaching, that lead towards their fate.
Saadalla Wannous is considered from the most important seekers for new technical forms in the Arabic Theatre. This appears through his plays that bear a collection of modern technical constituents. ...His early plays had technical structures that gave them technical and intellectual dimensions. The plays of the nineties were qualified with new tincture in both the structure and the meaning. From the technical constituents that formed this phase in writing the scripts is the narration technique that Wannous entered as a major structure in his drama, one of which is (Yawm Min Zamanina) (A Day from our Time) , in which he used diverse narrative techniquesto contributed in the production of the indicatives and signs and achieved a dramatic integration in the Play building up.
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