RUNX3 Regulates Intercellular Adhesion Molecule 3 (ICAM-3) Expression during Macrophage Differentiation and Monocyte Extravasation

The adhesion molecule ICAM-3 belongs to the immunoglobulin gene superfamily and functions as a ligand for the β2 integrins LFA-1, Mac-1 and αdβ2. The expression of ICAM-3 is restricted to cells of the hematopoietic lineage. We present evidences that the ICAM-3 gene promoter exhibits a leukocyte-specific activity, as its activity is significantly higher in ICAM-3+ hematopoietic cell lines. The activity of the ICAM-3 gene promoter is dependent on the occupancy of RUNX cognate sequences both in vitro and in vivo, and whose integrity is required for RUNX responsiveness and for the cooperative actions of RUNX with transcription factors of the Ets and C/EBP families. Protein analysis revealed that ICAM-3 levels diminish upon monocyte-derived macrophage differentiation, monocyte transendothelial migration and dendritic cell maturation, changes that correlate with an increase in RUNX3. Importantly, disruption of RUNX-binding sites led to enhanced promoter activity, and small interfering RNA-mediated reduction of RUNX3 expression resulted in increased ICAM-3 mRNA levels. Altogether these results indicate that the ICAM-3 gene promoter is negatively regulated by RUNX transcription factors, which contribute to the leukocyte-restricted and the regulated expression of ICAM-3 during monocyte-to-macrophage differentiation and monocyte extravasation

Transcriptomic analysis of the temporal host response to skin infestation with the ectoparasitic mite Psoroptes ovis

<p>Abstract</p> <p>Background</p> <p>Infestation of ovine skin with the ectoparasitic mite <it>Psoroptes ovis </it>results in a rapid cutaneous immune response, leading to the crusted skin lesions characteristic of sheep scab. Little is known regarding the mechanisms by which such a profound inflammatory response is instigated and to identify novel vaccine and drug targets a better understanding of the host-parasite relationship is essential. The main objective of this study was to perform a combined network and pathway analysis of the <it>in vivo </it>skin response to infestation with <it>P. ovis </it>to gain a clearer understanding of the mechanisms and signalling pathways involved.</p> <p>Results</p> <p>Infestation with <it>P. </it>ovis resulted in differential expression of 1,552 genes over a 24 hour time course. Clustering by peak gene expression enabled classification of genes into temporally related groupings. Network and pathway analysis of clusters identified key signalling pathways involved in the host response to infestation. The analysis implicated a number of genes with roles in allergy and inflammation, including pro-inflammatory cytokines (<it>IL1A, IL1B, IL6, IL8 </it>and <it>TNF</it>) and factors involved in immune cell activation and recruitment (<it>SELE, SELL, SELP, ICAM1, CSF2, CSF3, CCL2 </it>and <it>CXCL2</it>). The analysis also highlighted the influence of the transcription factors NF-kB and AP-1 in the early pro-inflammatory response, and demonstrated a bias towards a Th2 type immune response.</p> <p>Conclusions</p> <p>This study has provided novel insights into the signalling mechanisms leading to the development of a pro-inflammatory response in sheep scab, whilst providing crucial information regarding the nature of mite factors that may trigger this response. It has enabled the elucidation of the temporal patterns by which the immune system is regulated following exposure to <it>P. ovis</it>, providing novel insights into the mechanisms underlying lesion development. This study has improved our existing knowledge of the host response to <it>P. ovis</it>, including the identification of key parallels between sheep scab and other inflammatory skin disorders and the identification of potential targets for disease control.</p

Biosecurity Signalering 2014 : Bureau Biosecurity

In 2013 heeft de Nederlandse overheid Bureau Biosecurity opgericht als kennis- en informatiepunt voor zowel de overheid als voor organisaties die werken met risicovolle ziekteverwekkers. Biosecurity gaat over de beveiliging van deze ziekteverwekkers. Het werken in laboratoria met risicovolle ziekteverwekkers brengt risico's met zich mee, en het is daarom van belang dat in een laboratorium veilig met deze ziekteverwekkers wordt gewerkt - ter bescherming van mens en milieu (biosafety), maar ook beveiligd (biosecurity) - zodat het materiaal en laboratorium afgeschermd is voor kwaadwillenden. Bureau Biosecurity ontwikkelt kennisproducten, informeert over biosecurity en vergroot de bewustwording rondom biosecurity vraagstukken. Het bureau bundelt jaarlijks relevante nieuwsberichten en signalen over het onderwerp biosecurity. In de 'Biosecurity Signalering 2014' rapporteert Bureau Biosecurity over nationale en internationale biosecurity initiatieven, berichten vanuit de beleidshoek en wetenschappelijke publicaties die met biosecurity te maken hebben. Tevens zijn berichten uit de media opgenomen, zoals berichtgeving in kranten en internetartikelen gerelateerd aan biosecurity, die een afspiegeling vormen van de nieuwsgeving en perceptie in de samenleving. Het doel van de rapportage is om betrokken overheidspartijen over biosecurity-ontwikkelingen te informeren die van invloed kunnen zijn op risico's en dreigingen in Nederland. Hiertoe behoren ook rapportages over fouten die in laboratoria voorkomen en de kwetsbaarheden die hiermee aan het licht komen. Deze informatie kan van belang zijn voor hetbeleidsvormingstraject en het in kaart brengen van nieuwe risico's rondom risicovolle ziekteverwekkers. Bij de selectie van de berichten is gelet op de onderwerpen: biosecurity, dual-use onderzoek, gain-offunction onderzoek en biosafety- en biosecurity-incidenten met risicovolle ziekteverwekkers.In 2013, the Dutch government established the Netherlands Biosecurity Office as the national information centre for both the Dutch government and for organizations that work with high-risk pathogens. Laboratory work involving pathogens carries certain risks and therefore, personnel must work safely to protect the environment and human health ('biosafety'), but must also work in a secure manner to ensure that persons with malicious intent cannot gain access to the laboratory or the relevant biological materials or knowledge ('biosecurity'). The Biosecurity Office develops knowledge products, provides information, and increases awareness around biosecurity issues. Every year, the office compiles an overview of relevant news items and reports relating to biosecurity. In its Annual Biosecurity Overview 2014, the office reports on international biosecurity initiatives, policy developments, and scientific publications related to biosecurity. The Annual Biosecurity Overview also includes newspaper reports, internet articles and other media items relating to biosecurity that reflect news coverage and public perceptions of this topic. The purpose of the overview is to inform governmental stakeholders about biosecurity developments that may affect the risk and threat profile of the Netherlands. This includes reporting on errors occurring in laboratories and the vulnerabilities they reveal. This information can be useful to policy-makers, and can help in identifying new risks associated with pathogens. The news reports and articles were selected based on the following topics: biosecurity, dual use research, gain-of-function research, and biosafety and biosecurity incidents involving high-risk pathogens.Ministerie van VW

Antigen dose, type of antigen-presenting cell and time of differentiation contribute to the T helper 1/T helper 2 polarization of naive T cells

Antigenic encounter by T cells induces immunological synapse formation and T-cell activation. Using different concentrations of toxic shock syndrome toxin-1 (TSST-1) as stimulus, we examined the capacities of dendritic cells (DC) and macrophages (Mφ) to prime syngeneic naive T cells. DCs were, under all experimental settings, more efficient than Mφ at clustering T cells. Translocation of the T-cell receptor (TCR) to the contact area was found to be induced by DCs, as well as by Mφ, in an antigen-dependent manner, although Mφ were less efficient at inducing TCR translocation. Capping of protein kinase C θ (PKCθ) was also antigen dependent but induced exclusively by DCs. Likewise, DCs were found to be more potent inducers of interleukin-2 (IL-2) production and proliferation of naive T cells than Mφ. After 3 days of culture, DCs presenting 100 ng/ml TSST-1 induced interferon-γ (IFN-γ)-secreting cells, whereas Mφ did not. After 7 days of culture, DCs presenting 0·1 ng/ml TSST-1, and Mφ presenting high (as well as low) doses of TSST-1, induced IL-4-producing cells. We therefore provide evidence to show that antigen dose, type of antigen-presenting cell and time of differentiation can contribute to T-cell differentiation

Expression of ERBB3 binding protein 1 (EBP1) in salivary adenoid cystic carcinoma and its clinicopathological relevance

<p>Abstract</p> <p>Background</p> <p>ERBB3 binding protein 1 (<b><it>EBP1</it></b>) gene transfer into human salivary adenoid cystic carcinoma cells has been shown to significantly inhibit cell proliferation and reduce tumor metastasis in mouse models. In the current study, to evaluate if EBP1 is a novel biomarker capable of identifying patients at higher risk of disease progression and recurrence, we examined the EBP1 expression profile in adenoid cystic carcinoma (ACC) patients and analyzed its clinicopathological relevance. To understand the underlying anti-metastatic mechanism, we investigated if EBP1 regulates invasion-related molecules.</p> <p>Methods</p> <p>We performed immunohistochemical analysis on 132 primary adenoid cystic carcinoma and adjacent non-cancerous tissues using commercial EBP1, MMP9, E-cadherin and ICAM-1 antibodies. Results were correlated to clinicopathological parameters, long-term survival and invasion-related molecules by statistical analysis. Cell motility and invasiveness of vector or wild-type <it>EBP1</it>-transfected ACC-M cell lines were evaluated using wound healing and Boyden chamber assays. MMP9, E-cadherin and ICAM-1 proteins in these cell lines were detected using western blot assay.</p> <p>Results</p> <p>The expression of EBP1 was significantly higher in non-cancerous adjacent tissues compared with corresponding cancer tissues. The intensity and percentage of cells that reacted with EBP1 antibodies were significantly higher in cases with tubular pattern than those with solid pattern (<it>P</it><0.0001). We also found adenoid cystic carcinoma with local lymphatic metastasis had significantly lower EBP1 expression than ACC with no local lymphatic node metastasis (<it>P</it><0.0001). Similar findings were observed in ACC with lung metastasis compared with cases with no lung metastasis (<it>P</it><0.0001), in particular, in cases with perineural invasion compared with cases with no perineural invasion (<it>P</it><0.0001). Furthermore, a decrease in EBP1 expression was positively associated with a reduction in overall survival of ACC patients. Of note, EBP1 inhibits migration and invasiveness of ACC cells by upregulating E-cadherin but downregulating MMP9. In clinical adenoid cystic carcinoma patients, higher EBP1 expression was positively correlated with E-cadherin levels (<it>P</it><0.001) but negatively correlated with MMP9 expression (<it>P</it>=0.0002).</p> <p>Conclusions</p> <p>EBP1 expression is reduced in adenoid cystic carcinoma, indicating unfavorable prognosis of ACC patients. Its regulation of MMP9 and E-cadherin protein levels suggests a critical therapeutic potential.</p