ABSTRACT
The aim of this study was to examine the expression of the cytokines and chemokines receptor-3 (CCR3) molecule in endothelial cells and vascular structures in a murine model of corneal neovascularization and in samples of neovascularized human corneas. An immunofluorescence assay using the murine model showed a greater proportion and intensity of CCR3 in the epithelium and corneal subepithelial regions in corneas with neovascularization. In the absence of vascularization, no CCR3 was found. Of the 32 studied tissues, eight were vascularized and 24 were avascular. In the human corneas, vascularized corneas showed positive labeling for CD31 in all the analzedtissues, as well as positive labeling for CCR3. Therefore, all vascularized tissues showed positive coexpression of CCR3 and CD31, whereas none of the avascular corneas showed immunolabeling for either of these receptors. These results suggest that CCR3 could be a possible marker for corneal neovascularization with potential to be a therapeutic target.
Subject(s)
Cornea/metabolism , Corneal Neovascularization/genetics , Gene Expression Regulation , RNA/genetics , Receptors, CCR3/genetics , Animals , Cornea/pathology , Corneal Neovascularization/metabolism , Corneal Neovascularization/pathology , Disease Models, Animal , Humans , Immunohistochemistry , Mice , Mice, Inbred BALB C , Receptors, CCR3/biosynthesisABSTRACT
Allergic asthma is a chronic inflammatory disease characterized by the accumulation of eosinophils, Th2 cells and mononuclear cells in the airways, leading to changes in lung architecture and subsequently reduced respiratory function. We have previously demonstrated that CDIP-2, a chemokine derived peptide, reduced in vitro chemotaxis and decreased cellular infiltration in a murine model of allergic airway inflammation. However, the mechanisms involved in this process have not been identified yet. Now, we found that CDIP-2 reduces chemokine-mediated functions via interactions with CCR1, CCR2 and CCR3. Moreover, using bone marrow-derived eosinophils, we demonstrated that CDIP-2 modifies the calcium fluxes induced by CCL11 and down-modulated CCR3 expression. Finally, CDIP-2 treatment in a murine model of OVA-induced allergic airway inflammation reduced leukocyte recruitment and decreases production of cytokines. These data suggest that chemokine-derived peptides represent new therapeutic tools to generate more effective antiinflammatory drugs.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Peptides/pharmacology , Receptors, CCR1/metabolism , Receptors, CCR2/metabolism , Receptors, CCR3/metabolism , Allergens , Animals , Anti-Inflammatory Agents/therapeutic use , CHO Cells , Calcium/metabolism , Cell Line, Tumor , Chemotaxis/drug effects , Cricetulus , Cytokines/metabolism , Eosinophils/drug effects , Eosinophils/physiology , Female , Humans , Lung/drug effects , Lung/pathology , Lymph Nodes/cytology , Mice, Inbred BALB C , Ovalbumin , Peptides/therapeutic use , Pneumonia/drug therapy , Pneumonia/pathology , Receptors, CCR1/genetics , Receptors, CCR2/genetics , Receptors, CCR3/genetics , Respiratory Hypersensitivity/drug therapy , Respiratory Hypersensitivity/pathologyABSTRACT
PURPOSE: To compare gene expression of the chemokines RANTES and eotaxin-2, its receptor, CCR-3, adhesion molecule ICAM-1 and its receptor LFA-1 in eosinophilic polyps and in control normal nasal mucosa. METHODS: Gene expression was quantified by Real Time PCR in polyps (n=35) and in healthy nasal mucosa (n=15). RESULTS: Eosinophilic polyps showed a higher expression of eotaxin-2 and RANTES, but not of CCR-3, ICAM-1 or LFA-1 compared to control nasal mucosa. CONCLUSION: Eosinophilic polyps present greater expression of eotaxin-2 and RANTES, but not of CCR-3, ICAM-1 or LFA-1 compared to control nasal mucosa.
Subject(s)
Nasal Polyps/metabolism , Rhinitis/metabolism , Sinusitis/metabolism , Case-Control Studies , Chemokine CCL24/genetics , Chemokine CCL24/metabolism , Chemokine CCL5/genetics , Chemokine CCL5/metabolism , Chronic Disease , Gene Expression , Humans , Intercellular Adhesion Molecule-1/genetics , Intercellular Adhesion Molecule-1/metabolism , Lymphocyte Function-Associated Antigen-1/genetics , Lymphocyte Function-Associated Antigen-1/metabolism , Nasal Mucosa , Nasal Polyps/complications , Polymerase Chain Reaction , Receptors, CCR3/genetics , Receptors, CCR3/metabolism , Rhinitis/complications , Sinusitis/complicationsABSTRACT
BACKGROUND: The chemokine receptor CCR3 mediates the migration of cells that play an important role in the pathogenesis of asthma to inflammatory foci. Interferon (IFN)-gamma is known to downregulate the expression of some chemokine receptors. Therefore, we decided to analyze the regulation of CCR3 by IFN-gamma in asthmatics and to characterize the dependence of this process on immunoglobulin E (IgE) levels. METHODS: Atopic asthmatics were treated with IFN-gamma or placebo, and the IgE concentration in the blood was measured using an ultra-micro-ELISA for total IgE. Mononuclear cells from patients and controls were isolated by Ficoll-Hypaque gradient and incubated in the absence or presence of IFN-gamma for different periods of time. After incubation, the cells were washed and lysed for RT-PCR analysis, which was performed using a Perkin-Elmer kit. RESULTS: IFN-gamma treatment apparently improved the evaluated clinical variables; however, the differences were not significant compared to the placebo group. We found that IFN-gamma downregulated CCR3 mRNA expression ex vivo and in vivo in those patients with IgE levels higher than 500 IU/ml, whereas IFN-gamma upregulated CCR3 mRNA expression in patients with IgE levels lower than 500 IU/ml. Correspondence between ex vivo and in vivo results was observed using this approach. There was found to be a direct correlation between total serum IgE and CCR3 mRNA expression. CONCLUSIONS: In those asthmatic patients with high levels of IgE, who are thus susceptible to downregulation of CCR3 by IFN-gamma, a significant therapeutic effect with systemic IFN-gamma might be expected.