n-Nonanoyl-CC Chemokine Ligand 14, a Potent CC Chemokine Ligand 14 Analogue That Prevents the Recruitment of Eosinophils in Allergic Airway Inflammation

Abstract

CCR3 is responsible for tissue infiltration of eosinophils, basophils, mast cells, and Th2 cells, particularly in allergic diseases. In this context, CCR3 has emerged as a target for the treatment of allergic asthma. It is well known that the N-terminal domain of chemokines is crucial for receptor binding and, in particular, its activation. Based on this background, we investigated a number of N-terminally truncated or modified peptides derived from the chemokine CCL14/hemofiltrate CC chemokine-1 for their ability to modulate the activity of CCR3. Among 10 derivatives tested, n-nonanoyl (NNY)-CCL14[10–74] (NNY-CCL14) was the most potent at evoking the release of reactive oxygen species and inducing chemotaxis of human eosinophils. In contrast, NNY-CCL14 has inactivating properties on human eosinophils, because it is able to induce internalization of CCR3 and to desensitize CCR3-mediated intracellular calcium release and chemotaxis. In contrast to naturally occurring CCL11, NNY-CCL14 is resistant to degradation by CD26/dipeptidyl peptidase IV. Because inhibition of chemokine receptors through internalization is a reasonable therapeutic strategy being pursued for HIV infection, we tested a potential inhibitory effect of NNY-CCL14 in two murine models of allergic airway inflammation. In both OVA- and Aspergillus fumigatus-sensitized mice, i.v. treatment with NNY-CCL14 resulted in a significant reduction of eosinophils in the airways. Moreover, airway hyper-responsiveness was shown to be reduced by NNY-CCL14 in the OVA model. It therefore appears that an i.v. administered agonist internalizing and thereby inhibiting CCR3, such as NNY-CCL14, has the potential to alleviate CCR3-mediated diseases.

Recognition Versus Adaptive Up-regulation and Degradation of CC Chemokines by the Chemokine Decoy Receptor D6 Are Determined by Their N-terminal Sequence

“…In conclusion, results presented here indicate that D6 cooperates with CD26/dipeptidyl peptidase IV (29) in the inactivation and degradation of biologically active CCL14 isoforms. Moreover, by using CCL14 and other chemokine isoforms, we have shown that a proline residue in position 2 of D6 ligands is dispensable for receptor binding, but it is required to induce receptor traffic modifications resulting in efficient degradation of inflammatory CC chemokines.…”

“…Upon removal of the first 8 amino acid residues from the N terminus of CCL14 by urokinase plasminogen activator and/or plasmin, the prochemokine is converted into CCL14(9 -74), a potent agonist for CCR1, CCR3, and CCR5 (26 -28). Interestingly, the new N terminus is now recognized and processed by the dipeptidyl peptidase IV (CD26), which further cleaves 2 amino acids and generates the biologically inactive CCL14(11-74) variant (29).…”

Inhibition of CD26/Dipeptidyl Peptidase IV Enhances CCL11/Eotaxin-Mediated Recruitment of Eosinophils In Vivo

“…Human CCL11 was obtained from PeproTech and n-nonanoyl (NNY)-CCL14 was prepared as previously described (11). CCL11 was prepared as follows: CCL11 (0.5 mg) was dissolved in 1.5 ml of 0.01 M Tris-HCl buffer (pH 7.4).…”

“…After 1 and 24 h, samples of 20 or 50 l were taken and the degradation was stopped with 2 or 5 l of 2% TFA and frozen to Ϫ20°C immediately. Cleavage products of CCL11 were determined offline with a MALDI-MS (Voyager DE-Pro) in linear positive ion mode, as described recently (11).…”

Chemokines in tumor angiogenesis and metastasis

“…NNY-CCL14 (CCL14 analogue), CCL15, CCL24 and CCL26-28 [26, [54][55][56][57]. CCR4 is a receptor for CCL17 and CCL22 [58,59], whereas CCR5 is a major co-receptor for macrophage (M)-tropic HIV-1, HIV-2 and SIV strains [60].…”