title: Differences in potency of CXC chemokine ligand 8-, CC chemokine ligand 11-, and C5a-induced modulation of integrin function on human eosinophils.
abstract: The hypothesis was tested that different chemoattractants have different effects on the activity of integrins expressed by the human eosinophil. Three chemoattractants, CXCL8 (IL-8), CCL11 (eotaxin-1), and C5a were tested with respect to their ability to induce migration and the transition of eosinophils from a rolling interaction to a firm arrest on activated endothelial cells under flow conditions. CCL11 and C5a induced a firm arrest of eosinophils rolling on an endothelial surface, whereas CXCL8 induced only a transient arrest of the cells. The CXCL8- and CCL11-induced arrest was inhibited by simultaneously blocking alpha4 integrins (HP2/1) and beta2 integrins (IB4). In contrast, the C5a-induced arrest was only inhibited by 30% under these conditions. The potency differences of C5a>CCL11>CXCL8 to induce firm adhesion under flow condition was also observed in migration assays and for the activation of the small GTPase Rap-1, which is an important signaling molecule in the inside-out regulation of integrins. Interestingly, only C5a was able to induce the high activation epitope of alphaMbeta2 integrin recognized by MoAb CBRM1/5. The C5a-induced appearance of this epitope and Rap activation was controlled by phospholipase C (PLC), as was shown with the PLC inhibitor U73122. These data show that different chemoattractants are able to induce distinct activation states of integrins on eosinophils and that optimal chemotaxis is associated with the high activation epitope of the alphaMbeta2 integrin. Furthermore, PLC plays an important role in the inside-out signaling and, thus, the activation status of integrins on eosinophils.
authors.affiliation: Department of Pulmonary Diseases, University Medical Center Utrecht, Utrecht, The Netherlands. L.Ulfman@hli.azu.nl
authors.forename: Laurien H
authors.initials: LH
authors.lastname: Ulfman
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Alblas
authors.forename: Corneli W
authors.initials: CW
authors.lastname: van Aalst
authors.forename: Jaap Jan
authors.initials: JJ
authors.lastname: Zwaginga
authors.forename: Leo
authors.initials: L
authors.lastname: Koenderman
mesh_terms: D018821:CD18 Antigens / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects; D054413:Chemokine CCL11; D019742:Chemokines, CC / Q000494:pharmacology*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D015936:Complement C5a / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects; D004804:Eosinophils / Q000187:drug effects* / Q000276:immunology / Q000502:physiology; D006801:Humans; D039441:Integrin alpha4 / Q000502:physiology; D016023:Integrins / Q000502:physiology*; D016209:Interleukin-8 / Q000494:pharmacology*; D016177:Macrophage-1 Antigen / Q000502:physiology; D010738:Type C Phospholipases / Q000502:physiology; D020827:rap1 GTP-Binding Proteins / Q000502:physiology
chemical_list: C515456:CCL11 protein, human; D018821:CD18 Antigens; D054413:Chemokine CCL11; D019742:Chemokines, CC; D016023:Integrins; D016209:Interleukin-8; D016177:Macrophage-1 Antigen; D039441:Integrin alpha4; D015936:Complement C5a; D010738:Type C Phospholipases; D020827:rap1 GTP-Binding Proteins