IL-8 released constitutively by primary bronchial epithelial cells in culture forms an inactive complex with secretory component.
The bronchial epithelium is a source of both alpha and beta chemokines and, uniquely, of secretory component (SC), the extracellular ligand-binding domain of the polymeric IgA receptor. Ig superfamily relatives of SC, such as IgG and alpha(2)-macroglobulin, bind IL-8. Therefore, we tested the hypothesis that SC binds IL-8, modifying its activity as a neutrophil chemoattractant. Primary bronchial epithelial cells were cultured under conditions to optimize SC synthesis. The chemokines IL-8, epithelial neutrophil-activating peptide-78, growth-related oncogene alpha, and RANTES were released constitutively by epithelial cells from both normal and asthmatic donors and detected in high m.w. complexes with SC. There were no qualitative differences in the production of SC-chemokine complexes by epithelial cells from normal or asthmatic donors, and in all cases this was the only form of chemokine detected. SC contains 15% N-linked carbohydrate, and complete deglycosylation with peptide N-glycosidase F abolished IL-8 binding. In micro-Boyden chamber assays, no IL-8-dependent neutrophil chemotactic responses to epithelial culture supernatants could be demonstrated. SC dose-dependently (IC(50) approximately 0.3 nM) inhibited the neutrophil chemotactic response to rIL-8 (10 nM) in micro-Boyden chamber assays and also inhibited IL-8-mediated neutrophil transendothelial migration. SC inhibited the binding of IL-8 to nonspecific binding sites on polycarbonate filters and endothelial cell monolayers, and therefore the formation of haptotactic gradients, without effects on IL-8 binding to specific receptors on neutrophils. The data indicate that in the airways IL-8 may be solubilized and inactivated by binding to SC.
Marshall
L J
LJ
Department of Medical Specialities, Southampton General Hospital, Southampton, Hampshire, United Kingdom.
Perks
B
B
Ferkol
T
T
Shute
J K
JK
D001249:Asthma / Q000276:immunology; D001665:Binding Sites; D001980:Bronchi / Q000276:immunology*; D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis / Q000737:chemistry; D002634:Chemotaxis, Leukocyte; D004847:Epithelial Cells / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000737:chemistry; D046911:Macromolecular Substances; D008970:Molecular Weight; D009504:Neutrophils / Q000276:immunology; D012635:Secretory Component / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction
D018925:Chemokines; D016209:Interleukin-8; D046911:Macromolecular Substances; D012635:Secretory Component