IL-25 blockade augments antiviral immunity during respiratory virus infection.

IL-25 is implicated in the pathogenesis of viral asthma exacerbations. However, the effect of IL-25 on antiviral immunity has yet to be elucidated. We observed abundant expression and colocalization of IL-25 and IL-25 receptor at the apical surface of uninfected airway epithelial cells and rhinovirus infection increased IL-25 expression. Analysis of immune transcriptome of rhinovirus-infected differentiated asthmatic bronchial epithelial cells (BECs) treated with an anti-IL-25 monoclonal antibody (LNR125) revealed a re-calibrated response defined by increased type I/III IFN and reduced expression of type-2 immune genes CCL26, IL1RL1 and IL-25 receptor. LNR125 treatment also increased type I/III IFN expression by coronavirus infected BECs. Exogenous IL-25 treatment increased viral load with suppressed innate immunity. In vivo LNR125 treatment reduced IL-25/type 2 cytokine expression and increased IFN-ß expression and reduced lung viral load. We define a new immune-regulatory role for IL-25 that directly inhibits virus induced airway epithelial cell innate anti-viral immunity.

Inhibition of in vitro and in vivo T cell responses by recombinant human Tim-1 extracellular domain proteins.

Members of the T cell, Ig domain and mucin domain (Tim) family of proteins have recently been implicated in the control of T cell-mediated immune responses. Tim-1 (HUGO designation HAVCR1) polymorphisms have been linked to the regulation of atopy in mice and humans, suggestive of a role in immune regulation. Tim-1 is expressed upon activation of T cells. In concert with the increased expression of Tim-1, a binding partner for the extracellular domain of Tim-1 (eTim-1) was induced on activated T cells, and mRNA expression data was consistent with the binding partner being Tim-4. We found that co-immobilized recombinant eTim-1 was able to inhibit T cell activation mediated by CD3 + CD28 mAb. eTim-1 mediated its inhibitory effects on proliferation by arresting cell cycle at G(0)/G(1) phase through regulation of cell cycle proteins. In vivo, administration of eTim-1 proteins led to a decrease in both ear (contact hypersensitivity to oxazolone) and joint (methylated BSA antigen-induced arthritis) swelling. The inhibitory activity of eTim-1 in the T(h)1-dependent models was evidence that eTim-1 is able to modulate T cell responses. Manipulation of the Tim-1 interaction with its binding partner on T cells may therefore provide a novel target for therapeutic intervention in T cell-mediated diseases.

GeneCalling: transcript profiling coupled to a gene database query.

We describe here the GeneCalling method for the discovery of differentially expressed genes, both known and novel, from any species and with useful sequence information to determine the potential function of novel genes captured. The method relies on transcript visualization coupled to a database query to rapidly and quantitatively identify differentially expressed transcripts. The method has been applied to a wide variety of disease models in a wide variety of species, addressing problems as diverse as identifying novel human cancer gene targets, understanding how drugs and diet affect animal models of disease, and understanding the basis of trait differences in related strains of corn.

Gene expression quantitation technology summary.

Scientists routinely talk and write about gene expression and the abundance of transcripts, but in reality they extrapolate this information from the various measurements that a variety of different technologies provide. Indeed, there are many reasons why applying different technologies to the problem of transcript abundance may give different results, owing to an incomplete understanding of the gene in question or from shortcomings in the applications of the technologies. There are nine basic considerations for making a technology choice for quantitating gene expression that will impact the overall outcome: architecture, specificity, sensitivity, sample requirement, coverage, throughput, cost, reproducibility, and data management. These considerations will be discussed in the context of available technologies.

GeneCalling: transcript profiling coupled to a gene database query.

We describe the GeneCalling method for the discovery of differentially expressed genes, both known and novel, from any species including useful sequence information to determine the potential function of novel genes captured. The method relies on transcript visualization coupled to a database query to rapidly and quantitatively identify differentially expressed transcripts. The method has been applied to a wide variety of disease models in a variety of species, addressing problems as diverse as identifying novel human cancer gene targets, understanding how drugs and diet affect animal models of disease, and understanding the basis of trait differences in related strains of corn.

Angioarrestin: an antiangiogenic protein with tumor-inhibiting properties.

The angiopoietins comprise a family of proteins that have pro or antiangiogenic activities. Through a proprietary technology designed to identify transcripts of all expressed genes, we isolated a cDNA encoding an angiopoietin-related protein that we designate angioarrestin. The mRNA expression profile of angioarrestin was striking in that it was down-regulated in many tumor tissues when compared with adjacent nontumor tissue, suggesting a role for this protein in tumor inhibition. To test this hypothesis, we ectopically expressed angioarrestin in HT1080 tumor cells and measured pulmonary tumor nodule formation in nude mice. HT1080 cells expressing angioarrestin showed a marked reduction in the number and size of tumor nodules. In vitro, the recombinant protein was systematically tested in a number of endothelial cell assays and found to block critical processes involved in the angiogenic cascade, such as vascular endothelial growth factor/basic fibroblast growth factor-mediated endothelial cell proliferation, migration, tubular network formation, and adhesion to extracellular matrix proteins. These findings reveal a novel function for angioarrestin as an angiogenesis inhibitor and indicate that the molecule may be a potential cancer therapeutic.