title: The disulfide bond between cysteine 10 and cysteine 34 is required for CCL18 activity.
abstract: Asthma is a Th2-mediated disease that involves Th2 cell and eosinophil migration into the bronchial mucosa which is dependent upon the expression of a specific set of chemokines within the lung. Among them, CCL18 seems to play a key role because of its preferential expression in the lung, and its up-regulation by Th2 cytokines. Here, we show that the optimal naïve T cell and basophil chemotaxis, and basophil histamine release induced by rhCCL18 occurred at a 100 time lower concentration with CHO-derived rhCCL18 than with E. coli-derived rhCCL18. FT-ICR mass spectrometry of the intact chemokines showed that the rhCCL18 produced by CHO cells contained the 2 disulfide bonds Cys10-Cys34 and Cys11-Cys50, in clear contrast to the rhCCL18 derived from E. coli where the Cys10-Cys34 bond was absent. We found that reduction of the Cys10-Cys34 of the CHO-derived rhCCL18 resulted in a shift of its activity, reaching the same level as the E. coli-derived rhCCL18. These results demonstrate that the Cys10-Cys34 disulfide bond is involved in the function of CCL18.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, U1019, Pulmonary Immunity team, F-59019 Lille, France; Institut Pasteur de Lille, Center for Infection and Immunity of Lille, F-59019 Lille, France.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Legendre
authors.forename: Caroline
authors.initials: C
authors.lastname: Tokarski
authors.forename: Ying
authors.initials: Y
authors.lastname: Chang
authors.forename: Nathalie
authors.initials: N
authors.lastname: De Freitas Caires
authors.forename: Hugues
authors.initials: H
authors.lastname: Lortat-Jacob
authors.forename: Patricia De
authors.initials: PD
authors.lastname: Nadaï
authors.forename: Christian
authors.initials: C
authors.lastname: Rolando
authors.forename: Catherine
authors.initials: C
authors.lastname: Duez
authors.forename: Anne
authors.initials: A
authors.lastname: Tsicopoulos
authors.forename: Philippe
authors.initials: P
authors.lastname: Lassalle
mesh_terms: D000818:Animals; D001249:Asthma / Q000276:immunology / Q000378:metabolism*; D001491:Basophils / Q000276:immunology / Q000378:metabolism; D016466:CHO Cells; D002460:Cell Line; D002465:Cell Movement / Q000276:immunology; D019742:Chemokines, CC / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003412:Cricetulus; D003545:Cysteine / Q000737:chemistry* / Q000235:genetics; D004804:Eosinophils / Q000378:metabolism; D006632:Histamine / Q000276:immunology; D006636:Histamine Release; D006801:Humans; D008168:Lung / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology*
chemical_list: C111792:CCL18 protein, human; D019742:Chemokines, CC; D006632:Histamine; D003545:Cysteine
keywords: CCL18 activity; Disulfide bond; Intact protein analysis