Glycosaminoglycan Interactions with Chemokines Add Complexity to a Complex System.

Chemokines have two types of interactions that function cooperatively to control cell migration. Chemokine receptors on migrating cells integrate signals initiated upon chemokine binding to promote cell movement. Interactions with glycosaminoglycans (GAGs) localize chemokines on and near cell surfaces and the extracellular matrix to provide direction to the cell movement.

Antibody Neutralization of CXCL10 Is Dependent on Binding to Free and Not Endothelial-bound Chemokine: IMPLICATIONS FOR THE DESIGN OF A NEW GENERATION OF ANTI-CHEMOKINE THERAPEUTIC ANTIBODIES.

To improve our understanding of properties that confer successful inhibition of chemokines , we analyzed anti-murine CXCL10 monoclonal antibodies (mAb) having different characteristics. 1B6 displayed potent inhibition of cell recruitment with an IC of 0.5 nm but demonstrated little efficacy in various animal models of human disease.

Evasins: Therapeutic Potential of a New Family of Chemokine-Binding Proteins from Ticks.

Blood-sucking parasites, such as ticks, remain attached to their hosts for relatively long periods of time in order to obtain their blood meal without eliciting an immune response. One mechanism used to avoid rejection is the inhibition of the recruitment of immune cells, which can be achieved by a class of chemokine-binding proteins (CKBPs) known as Evasins. We have identified three distinct Evasins produced by the salivary glands of the common brown dog tick, Rhipicephalus sanguineus.

Mutagenesis by Phage Display.

Chemokines are small chemoattractant proteins involved in the recruitment of leukocytes to the site of inflammation. Due to their prominent role in the inflammatory process, chemokine inhibitors have been developed by parasites to remain undetected not only by the host immune system but also by various laboratories to develop anti-inflammatory compounds. Taking advantage of the small size of natural chemokine-binding proteins, we report here several methods to facilitate their characterization using phage display to identify the chemokine-binding site and to modulate the selectivity of such inhibitors.

Methods for the Recognition of GAG-Bound Chemokines.

Chemokines play a pivotal role in the multistep cascade of cellular recruitment, where they provide the directional signal. They activate cells through a high-affinity interaction with their receptors, members of the large family of heptahelical G protein-coupled receptors. In order to provide the directional signal, they bind to cell surface proteoglycans through a low-affinity interaction with the glycosaminoglycan (GAG) moiety.

Targeting chemokines: Pathogens can, why can't we?

Chemoattractant cytokines, or chemokines, are the largest sub-family of cytokines. About 50 distinct chemokines have been identified in humans. Their principal role is to stimulate the directional migration of leukocytes, which they achieve through activation of their receptors, following immobilization on cell surface glycosaminoglycans (GAGs).

De novo isolation of antibodies with pH-dependent binding properties.

pH-dependent antibodies are engineered to release their target at a slightly acidic pH, a property making them suitable for clinical as well as biotechnological applications. Such antibodies were previously obtained by histidine scanning of pre-existing antibodies, a labor-intensive strategy resulting in antibodies that displayed residual binding to their target at pH 6.0.

Identification of the pharmacophore of the CC chemokine-binding proteins Evasin-1 and -4 using phage display.

To elucidate the ligand-binding surface of the CC chemokine-binding proteins Evasin-1 and Evasin-4, produced by the tick Rhipicephalus sanguineus, we sought to identify the key determinants responsible for their different chemokine selectivities by expressing Evasin mutants using phage display. We first designed alanine mutants based on the Evasin-1·CCL3 complex structure and an in silico model of Evasin-4 bound to CCL3. The mutants were displayed on M13 phage particles, and binding to chemokine was assessed by ELISA.

CCL18 exhibits a regulatory role through inhibition of receptor and glycosaminoglycan binding.

CCL18 has been reported to be present constitutively at high levels in the circulation, and is further elevated during inflammatory diseases. Since it is a rather poor chemoattractant, we wondered if it may have a regulatory role. CCL18 has been reported to inhibit cellular recruitment mediated by CCR3, and we have shown that whilst it is a competitive functional antagonist as assessed by Schild plot analysis, it only binds to a subset of CCR3 receptor populations.

Evasin-4, a tick-derived chemokine-binding protein with broad selectivity can be modified for use in preclinical disease models.

Rhipicephalus sanguineus, the common brown dog tick, produces several chemokine-binding proteins which are secreted into the host in its saliva to modulate the host response during feeding. Two of these demonstrate very restricted selectivity profiles. Here, we describe the characterization of the third, which we named Evasin-4.

Dynamic octopus amphiphiles as powerful activators of DNA transporters: differential fragrance sensing and beyond.

We report the design, synthesis and evaluation of dynamic "octopus" amphiphiles with emphasis on their efficiency as activators in synthetic membrane-based sensing systems. Previously, we found that the in situ treatment of charged hydrazides with hydrophobic aldehydes or ketones gives amphiphilic counterion activators of polyion transporters in lipid bilayers, and that their efficiency increases with the number of their hydrophobic tails. Herein, we expand this series to amphiphiles with one cationic head (guanidinium or ammonium) and four exchangeable hydrophobic tails.