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lüll Novel association between vasoactive intestinal peptide and CRTH2 receptor in recruiting eosinophils: a possible biochemical mechanism for allergic eosinophilic inflammation of the airways El-Shazly AE; Begon DY; Kustermans G; Arafa M; Dortu E; Henket M; Lefebvre PP; Louis R; Delvenne PJ Biol Chem 2013[Jan]; 288 (2): 1374-84We explored the relation between vasoactive intestinal peptide (VIP), CRTH2, and eosinophil recruitment. It is shown that CRTH2 expression by eosinophils from allergic rhinitis (AR) patients and eosinophil cell line (Eol-1 cells) was up-regulated by VIP treatment. This was functional and resulted in exaggerated migratory response of cells against PGD2. Nasal challenge of AR patients resulted in a significant increase of VIP contents in nasal secretion (ELISA), and the immunohistochemical studies of allergic nasal tissues showed significant expression of VIP in association with intense eosinophil recruitment. Biochemical assays showed that VIP-induced eosinophil chemotaxis from AR patients and Eol-1 cells was mediated through the CRTH2 receptor. Cell migration against VIP was sensitive to protein kinase C (PKC) and protein kinase A (PKA) inhibition but not to tyrosine kinase or p38 MAPK inhibition or calcium chelation. Western blot demonstrated a novel CRTH2-mediated cytosol-to-membrane translocation of PKC-epsilon, PKC-delta, and PKA-alpha, -gamma, and -IIalphareg in Eol-1 cells upon stimulation with VIP. Confocal images and FACS demonstrated a strong association and co-localization between VIP peptide and CRTH2 molecules. Further, VIP induced PGD2 secretion from eosinophils. Our results demonstrate the first evidence of association between VIP and CRTH2 in recruiting eosinophils.|Amino Acid Sequence[MESH]|Base Sequence[MESH]|DNA Primers[MESH]|Enzyme-Linked Immunosorbent Assay[MESH]|Eosinophilia/metabolism/*pathology[MESH]|Eosinophils/*cytology[MESH]|Flow Cytometry[MESH]|Humans[MESH]|Hypersensitivity/metabolism/*pathology[MESH]|Immunohistochemistry[MESH]|Molecular Sequence Data[MESH]|Receptors, Immunologic/*metabolism[MESH]|Receptors, Prostaglandin/*metabolism[MESH]|Reverse Transcriptase Polymerase Chain Reaction[MESH]|Trachea/metabolism/*pathology[MESH]|Vasoactive Intestinal Peptide/chemistry/*metabolism[MESH] |