Transcription factors RUNX1 and RUNX3 in the induction and suppressive function of Foxp3+ inducible regulatory T cells

Forkhead box P3 (FOXP3)+CD4+CD25+ inducible regulatory T (iT reg) cells play an important role in immune tolerance and homeostasis. In this study, we show that the transforming growth factor-β (TGF-β) induces the expression of the Runt-related transcription factors RUNX1 and RUNX3 in CD4+ T cells. This induction seems to be a prerequisite for the binding of RUNX1 and RUNX3 to three putative RUNX binding sites in the FOXP3 promoter. Inactivation of the gene encoding RUNX cofactor core-binding factor-β (CBFβ) in mice and small interfering RNA (siRNA)-mediated suppression of RUNX1 and RUNX3 in human T cells resulted in reduced expression of Foxp3. The in vivo conversion of naive CD4+ T cells into Foxp3+ iT reg cells was significantly decreased in adoptively transferred CbfbF/F CD4-cre naive T cells into Rag2−/− mice. Both RUNX1 and RUNX3 siRNA silenced human T reg cells and CbfbF/F CD4-cre mouse T reg cells showed diminished suppressive function in vitro. Circulating human CD4+ CD25high CD127− T reg cells significantly expressed higher levels of RUNX3, FOXP3, and TGF-β mRNA compared with CD4+CD25− cells. Furthermore, FOXP3 and RUNX3 were colocalized in human tonsil T reg cells. These data demonstrate Runx transcription factors as a molecular link in TGF-β–induced Foxp3 expression in iT reg cell differentiation and function

Methylation matters: binding of Ets-1 to the demethylated Foxp3 gene contributes to the stabilization of Foxp3 expression in regulatory T cells

The forkhead-box protein P3 (Foxp3) is a key transcription factor for the development and suppressive activity of regulatory T cells (Tregs), a T cell subset critically involved in the maintenance of self-tolerance and prevention of over-shooting immune responses. However, the transcriptional regulation of Foxp3 expression remains incompletely understood. We have previously shown that epigenetic modifications in the CpG-rich Treg-specific demethylated region (TSDR) in the Foxp3 locus are associated with stable Foxp3 expression. We now demonstrate that the methylation state of the CpG motifs within the TSDR controls its transcriptional activity rather than a Treg-specific transcription factor network. By systematically mutating every CpG motif within the TSDR, we could identify four CpG motifs, which are critically determining the transcriptional activity of the TSDR and which serve as binding sites for essential transcription factors, such as CREB/ATF and NF-κB, which have previously been shown to bind to this element. The transcription factor Ets-1 was here identified as an additional molecular player that specifically binds to the TSDR in a demethylation-dependent manner in vitro. Disruption of the Ets-1 binding sites within the TSDR drastically reduced its transcriptional enhancer activity. In addition, we found Ets-1 bound to the demethylated TSDR in ex vivo isolated Tregs, but not to the methylated TSDR in conventional CD4+ T cells. We therefore propose that Ets-1 is part of a larger protein complex, which binds to the TSDR only in its demethylated state, thereby restricting stable Foxp3 expression to the Treg lineage

Experimental determination of the permeability of engineering textiles: Benchmark II

In this second international permeability benchmark, the in-plane permeability values of a carbon fabric were studied by twelve research groups worldwide. One participant also investigated the deformation of the tested carbon fabric. The aim of this work was to obtain comparable results in order to make a step toward standardization of permeability measurements. Unidirectional injections were thus conducted to determine the unsaturated in-plane permeability tensor of the fabric. Procedures used by participants were specified in the guidelines defined for this benchmark. Participants were asked to use the same values for parameters such as fiber volume fraction, injection pressure and fluid viscosity to minimize sources of scatter. The comparison of the results from each participant was encouraging. The scatter between data obtained while respecting the guidelines was below 25%. However, a higher dispersion was observed when some parameters differed from the recommendations of this exercise.The authors are grateful to J.M. Beraud from Hexcel Fabrics for his support that made possible this exercise. The contributions of J.B. Alms, N.C. Correia, S. Advani, E. Ruiz and P.C.T. Goncalves to the preparation of the guidelines document and templates are acknowledged by the participants of this benchmark.Vernet, N.; Ruiz, E.; Advani, S.; Alms, JB.; Aubert, M.; Barburski, M.; Barari, B.... (2014). Experimental determination of the permeability of engineering textiles: Benchmark II. Composites Part A: Applied Science and Manufacturing. 61:172-184. doi:10.1016/j.compositesa.2014.02.010S1721846

Genetic and transcriptomic analysis of transcription factor genes in the model halophilic Archaeon: coordinate action of TbpD and TfbA

<p>Abstract</p> <p>Background</p> <p>Archaea are prokaryotic organisms with simplified versions of eukaryotic transcription systems. Genes coding for the general transcription factors TBP and TFB are present in multiple copies in several Archaea, including <it>Halobacterium </it>sp. NRC-1. Multiple TBP and TFBs have been proposed to participate in transcription of genes via recognition and recruitment of RNA polymerase to different classes of promoters.</p> <p>Results</p> <p>We attempted to knock out all six TBP and seven TFB genes in <it>Halobacterium </it>sp. NRC-1 using the <it>ura</it>3-based gene deletion system. Knockouts were obtained for six out of thirteen genes, <it>tbp</it>CDF and <it>tfb</it>ACG, indicating that they are not essential for cell viability under standard conditions. Screening of a population of 1,000 candidate mutants showed that genes which did not yield mutants contained less that 0.1% knockouts, strongly suggesting that they are essential. The transcriptomes of two mutants, Δ<it>tbp</it>D and Δ<it>tfb</it>A, were compared to the parental strain and showed coordinate down regulation of many genes. Over 500 out of 2,677 total genes were regulated in the Δ<it>tbp</it>D and Δ<it>tfb</it>A mutants with 363 regulated in both, indicating that over 10% of genes in both strains require the action of both TbpD and TfbA for normal transcription. Culturing studies on the Δ<it>tbp</it>D and Δ<it>tfb</it>A mutant strains showed them to grow more slowly than the wild-type at an elevated temperature, 49°C, and they showed reduced viability at 56°C, suggesting TbpD and TfbA are involved in the heat shock response. Alignment of TBP and TFB protein sequences suggested the expansion of the TBP gene family, especially in <it>Halobacterium </it>sp. NRC-1, and TFB gene family in representatives of five different genera of haloarchaea in which genome sequences are available.</p> <p>Conclusion</p> <p>Six of thirteen TBP and TFB genes of <it>Halobacterium </it>sp. NRC-1 are non-essential under standard growth conditions. TbpD and TfbA coordinate the expression of over 10% of the genes in the NRC-1 genome. The Δ<it>tbp</it>D and Δ<it>tfb</it>A mutant strains are temperature sensitive, possibly as a result of down regulation of heat shock genes. Sequence alignments suggest the existence of several families of TBP and TFB transcription factors in <it>Halobacterium </it>which may function in transcription of different classes of genes.</p

The role of epigenetic dysregulation in the epidemic of allergic disease

The epidemic of allergic disease in early life is one of the clearest indicators that the developing immune system is vulnerable to modern environmental changes. A range of environmental exposures epidemiologically associated with allergic disease have been shown to have effects on the foetal immune function in pregnancy, including microbial burden, dietary changes and environmental pollutants. Preliminary studies now suggest that these early effects on immune development may be mediated epigenetically through a variety of processes that collectively modify gene expression and allergic susceptibility and that these effects are potentially heritable across generations. It is also possible that rising rates of maternal allergy, a recognised direct risk factor for infant allergic disease, may be further amplifying the effects of environmental changes. Whilst effective prevention strategies are the ultimate goal in reversing the allergy epidemic, the specific environmental drivers, target genes, and intracellular pathways and mechanisms of early life immune programming are still unclear. It is hoped that identifying genes that are differentially regulated in association with subsequent allergic disease will assist in identifying causal pathways and upstream contributing environmental factors. In this way, epigenetic paradigms are likely to provide valuable insights into how the early environment can be modified to more favourably drive immune development and reverse the allergic epidemic

Thermoplastic Protein Nano-Composites Using Bloodmeal and Bentonite

Bloodmeal is a low value meat industry product and can be converted into a thermoplastic material. These novel thermoplastics often have inferior mechanical properties and require some degree of reinforcement. Reinforcement using octadecylamine (OAmine) modified bentonite have shown an increase in tensile strength from 7.69 to 9.26 MPa by using 2 parts clay per hundred parts bloodmeal. Unmodified clay did not lead to the same increase in strength. High shear during extrusion was not sufficient to prevent agglomeration at higher clay content leading to modified clay composites showing reduced tensile strength. Particle size of unmodified clay composites was small enough to lead to dispersion reinforcement, whereas modified clay composites showed even greater agglomeration, leading to a decrease in strength. It was concluded that the physical nature of the thermoplastic protein requires careful consideration as to the level of clay modification required

Molekular-zytogenetische Analysen an alter DNA (aDNA) anhand von Praeparaten der Meckelschen Sammlungen zu Halle (Saale) [Molecular-cytogenetic analysis of ancient DNA (aDNA) from preparations from the Meckel collection in Halle (Saale)]

In this report, we present first results of molecular cytogenetic analyses by comparative genomic hybridization (CGH) on ancient DNA (aDNA) of two newborns preserved in the Meckel Collections. The goal of the analyses was the exclusion of a chromosomal imbalance accounting for the described malformations of the individuals. Comparative genomic hybridization (CGH) is a well proven molecular cytogenetic approach for genome-wide analysis of chromosomal gains and losses in DNA samples without preparing chromosomes of the test sample. Ancient DNA was extracted from the remaining umbilical cord, labeled and investigated for chromosomal imbalances by CGH. Both genomic aDNA probes showed a normal copy number karyotype. Additional molecular genetic experiments on these aDNAs were tested to follow up the question whether different other genetic investigations are possible on further samples of the Meckel Collections