Analysis of the CCR3 promoter reveals a regulatory region in exon 1 that binds GATA-1

BACKGROUND: CC Chemokine Receptor 3 (CCR3), the major chemokine receptor expressed on eosinophils, binds promiscuously to several ligands including eotaxins 1, 2, and 3. Even though the only cells that consistently accumulate following eotaxin administration in vivo are myeloid cells (primarily eosinophils), other cell types have recently been shown to express CCR3. It is therefore important to elucidate the molecular mechanisms regulating receptor expression. RESULTS: In order to define regions responsible for CCR3 transcription, a DNAse hypersensitive site was identified in the vicinity of exon 1. Coupled with our previous data implicating exon 1 in CCR3 transcription, we hypothesized that transcription factors bind to exon-1. Electrophoretic mobility shift analysis revealed that nuclear proteins in eosinophilic cells bound to exon 1. Furthermore, antibody interference and mutation studies demonstrated GATA-1 binding to exon 1. In order to test the 1.6-kb CCR3 promoter element (that includes exon 1) for in vivo function, this region was used to generate transgenic mice that expressed a reporter protein. Strong transgene expression was achieved, with the pattern of expression suggesting a broad acting promoter. CONCLUSION: The transcription factor GATA-1 binds to CCR3 exon 1. The 1.6-kb CCR3 promoter element, that includes exon 1, is a strong promoter in vivo

Bone marrow cell derived arginase I is the major source of allergen-induced lung arginase but is not required for airway hyperresponsiveness, remodeling and lung inflammatory responses in mice

<p>Abstract</p> <p>Background</p> <p>Arginase is significantly upregulated in the lungs in murine models of asthma, as well as in human asthma, but its role in allergic airway inflammation has not been fully elucidated in mice.</p> <p>Results</p> <p>In order to test the hypothesis that arginase has a role in allergic airway inflammation we generated arginase I-deficient bone marrow (BM) chimeric mice. Following transfer of arginase I-deficient BM into irradiated recipient mice, arginase I expression was not required for hematopoietic reconstitution and baseline immunity. Arginase I deficiency in bone marrow-derived cells decreased allergen-induced lung arginase by 85.8 ± 5.6%. In contrast, arginase II-deficient mice had increased lung arginase activity following allergen challenge to a similar level to wild type mice. BM-derived arginase I was not required for allergen-elicited sensitization, recruitment of inflammatory cells in the lung, and proliferation of cells. Furthermore, allergen-induced airway hyperresponsiveness and collagen deposition were similar in arginase-deficient and wild type mice. Additionally, arginase II-deficient mice respond similarly to their control wild type mice with allergen-induced inflammation, airway hyperresponsiveness, proliferation and collagen deposition.</p> <p>Conclusion</p> <p>Bone marrow cell derived arginase I is the predominant source of allergen-induced lung arginase but is not required for allergen-induced inflammation, airway hyperresponsiveness or collagen deposition.</p

Increased susceptibility of 129SvEvBrd mice to IgE-Mast cell mediated anaphylaxis

<p>Abstract</p> <p>Background</p> <p>Experimental analyses have identified strain-dependent factors that regulate susceptibility to anaphylaxis in mice. We assessed the susceptibility of the widely used 129SvEvBrd (also known as 129S5) mouse strain to IgE/mast cell-mediated anaphylaxis as compared to BALB/c. Mice were subjected to passive and oral Ovalbumin [OVA]-induced active anaphylaxis. Tissue mast cell, plasma histamine, total IgE and OVA-specific IgE levels and susceptibility to histamine i.v infusion were assessed. Bone marrow mast cell (BMMC)s were examined for Fc_εRI, c-kit, degranulation efficiency, proliferation, apoptosis and cytokine profile.</p> <p>Results</p> <p>129S5 mice had significantly increased susceptibility to passive and oral OVA-induced active anaphylaxis. Increased susceptibility to anaphylaxis was associated with increased homeostatic mast cell levels but not OVA-specific IgE or IgG₁levels. <it>In vitro </it>analyses of BMMCs revealed no difference in Fc_εRI and c-Kit expression, however, 129S5 BMMCs possessed greater proliferative capacity and reduced caspase-3-mediated apoptosis. IgE-BMMC degranulation assays demonstrated no difference in degranulation efficiency. Furthermore, 129S5 mice possessed increased sensitivity to histamine-induced hypothermia.</p> <p>Conclusions</p> <p>We conclude that 129S5 mice have increased susceptibility to anaphylaxis as compared to BALB/c strain and their increased susceptibility was associated with altered mast cell proliferation and homeostatic tissue levels and responsiveness to histamine. Given the wide spread usage of the 129SvEvBrd strain of mice in experimental gene targeting methodology, these data have important implications for studying IgE-reactions in mouse systems.</p

Prerequisites for coexistence: human pressure and refuge habitat availability shape continental‑scale habitat use patterns of a large carnivore

Context Adjustments in habitat use by large carnivores can be a key factor facilitating their coexistence with people in shared landscapes. Landscape composition might be a key factor determining how large carnivores can adapt to occurring alongside humans, yet broad-scale analyses investigating adjustments of habitat use across large gradients of human pressure and landscape composition are lacking. Objectives Here, we investigate adjustments in habitat use by Eurasian lynx (Lynx lynx) in response to varying availability of refuge habitats (i.e., forests and rugged terrain) and human landscape modifcation. Methods Using a large tracking dataset including 434 individuals from seven populations, we assess functional responses in lynx habitat use across two spatial scales, testing for variation by sex, daytime, and season. Results We found that lynx use refuge habitats more intensively with increasing landscape modifcation across spatial scales, selecting forests most strongly in otherwise open landscapes and rugged terrain in mountainous regions. Moreover, higher forest availability enabled lynx to place their home ranges in more human-modifed landscapes. Human pressure and refuge habitat availability also shaped temporal patterns of lynx habitat use, with lynx increasing refuge habitat use and reducing their use of human-modifed areas during periods of high exposure (daytime) or high vulnerability (postnatal period) to human pressure. Conclusions Our fndings suggest a remarkable adaptive capacity of lynx towards human pressure and underline the importance of refuge habitats across scales for enabling coexistence between large carnivores and people. More broadly, we highlight that the composition of landscapes determines how large carnivores can adapt to human pressure and thus play an important role shaping large carnivore habitat use and distributions.publishedVersio

Integrating animal tracking datasets at a continental scale for mapping Eurasian lynx habitat

Aim: The increasing availability of animal tracking datasets collected across many sites provides new opportunities to move beyond local assessments to enable de-tailed and consistent habitat mapping at biogeographical scales. However, integrating wildlife datasets across large areas and study sites is challenging, as species' varying responses to different environmental contexts must be reconciled. Here, we compare approaches for large-area habitat mapping and assess available habitat for a recolo-nizing large carnivore, the Eurasian lynx (Lynx lynx).Location: Europe.Methods: We use a continental-scale animal tracking database (450 individuals from 14 study sites) to systematically assess modelling approaches, comparing (1) global strategies that pool all data for training versus building local, site-specific models and combining them, (2) different approaches for incorporating regional variation in habi-tat selection and (3) different modelling algorithms, testing nonlinear mixed effects models as well as machine-learning algorithms.Results: Testing models on training sites and simulating model transfers, global and local modelling strategies achieved overall similar predictive performance. Model performance was the highest using flexible machine-learning algorithms and when incorporating variation in habitat selection as a function of environmental variation. Our best-performing model used a weighted combination of local, site-specific habi-tat models. Our habitat maps identified large areas of suitable, but currently unoccu-pied lynx habitat, with many of the most suitable unoccupied areas located in regions that could foster connectivity between currently isolated populations.Main Conclusions: We demonstrate that global and local modelling strategies can achieve robust habitat models at the continental scale and that considering regional variation in habitat selection improves broad-scale habitat mapping. More generally, we highlight the promise of large wildlife tracking databases for large-area habitat mapping. Our maps provide the first high-resolution, yet continental assessment of lynx habitat across Europe, providing a consistent basis for conservation planning for restoring the species within its former range.publishedVersio

Chemokines in asthma: Cooperative interaction between chemokines and IL-13

The asthmatic response is characterized by elevated production of IgE, cytokines, chemokines, mucus hypersecretion, airway obstruction, eosinophilia, and enhanced airway hyperreactivity to spasmogens. Clinical and experimental investigations have demonstrated a strong correlation between the presence of CD4+ TH2 cells, eosinophils, and disease severity, suggesting an integral role for these cells in the pathophysiology of asthma. TH2 cells are thought to induce asthma through the secretion of an array of cytokines (IL-4, -5, -9 -10, -13, -25) that activate inflammatory and residential effector pathways both directly and indirectly. In particular, IL-4 and IL-13 are produced at elevated levels in the asthmatic lung and are thought to be central regulators of many of the hallmark features of the disease. The potency of IL-13 in promoting airway hyperreactivity and mucus hypersecretion and the ability of IL-13 blockade to abrogate several critical aspects of experimental asthma have led to the view that this is a critical cytokine in disease pathogenesis. Extensive studies have also demonstrated a central role for chemokines in orchestrating multiple aspects of the asthmatic response. Chemokines are potent leukocyte chemoattractants, cellular activating factors, and histamine-releasing factors, which makes them particularly important in the pathogenesis of allergic inflammation. In particular, the eotaxin subfamily of chemokines and their receptor CC chemokine receptor 3 have emerged as central regulators of the asthmatic response. Recent studies have provided an integrated mechanism by which to explain the coordinate interaction between IL-13 and chemokines in the pathogenesis of asthma. In this regard, chemokines and IL-13 are attractive new therapeutic targets for the treatment of allergic disease. This article focuses on recently emerging data pertaining to the importance of chemokines, especially eotaxins, in promoting IL-13-associated allergic lung responses, as well as the potential for pharmacologically targeting these pathways

The cationic amino acid transporter 2 is induced in inflammatory lung models and regulates lung fibrosis

Abstract Background Arginine is an amino acid that serves as a substrate for the enzymes nitric oxide synthase (NOS) and arginase, leading to synthesis of NO and ornithine, respectively. As such, arginine has the potential to influence diverse fundamental processes in the lung. Methods We used mice deficient in cationic amino acid transporter (CAT) 2 in models of allergic airway inflammation and pulmonary fibrosis. Results We report that the arginine transport protein CAT2 was over-expressed in the lung during the induction of allergic airway inflammation. Furthermore, CAT2 mRNA was strongly induced by transgenically over-expressed IL-4, and allergen-induced expression was dependent upon signal-transducer-and-activator-of-transcription (STAT) 6. In situ mRNA hybridization demonstrated marked staining of CAT2, predominantly in scattered mononuclear cells. Analysis of allergic airway inflammation and bleomycin-induced inflammation in CAT2-deficient mice revealed that while inflammation was independent of CAT2 expression, bleomycin-induced fibrosis was dependent upon CAT2. Mechanistic analysis revealed that arginase activity in macrophages was partly dependent on CAT2. Conclusion Taken together, these results identify CAT2 as a regulator of fibrotic responses in the lung