title: Eotaxin is specifically cleaved by hookworm metalloproteases preventing its action in vitro and in vivo.
abstract: Eotaxin is a potent eosinophil chemoattractant that acts selectively through CCR3, which is expressed on eosinophils, basophils, mast cells, and Th2-type T cells. This arm of the immune system is believed to have evolved to control helminthic parasites. We hypothesized that helminths may employ mechanisms to inhibit eosinophil recruitment, to prolong worm survival in the host. We observed that the excretory/secretory products of the hookworm Necator americanus inhibited eosinophil recruitment in vivo in response to eotaxin, but not leukotriene B(4), a phenomenon that could be prevented by the addition of protease inhibitors. Using Western blotting, N. americanus supernatant was shown to cause rapid proteolysis of eotaxin, but not IL-8 or eotaxin-2. N. americanus homogenate was fractionated by gel filtration chromatography, and a FACS-based bioassay measured the ability of each fraction to inhibit the activity of a variety of chemokines. This resulted in two peaks of eotaxin-degrading activity, corresponding to approximately 15 and 50 kDa molecular mass. This activity was specific for eotaxin, as responses to other agonists tested were unaffected. Proteolysis of eotaxin was prevented by EDTA and phenanthroline, indicating that metalloprotease activity was involved. Production of enzymes inactivating eotaxin may be a strategy employed by helminths to prevent recruitment and activation of eosinophils at the site of infection. As such this represents a novel mechanism of regulation of chemokine function in vivo. The existence of CCR3 ligands other than eotaxin (e.g., eotaxin-2) may reflect the evolution of host counter measures to parasite defense systems.
authors.affiliation: Leukocyte Biology Section, Biomedical Sciences Division, Imperial College School of Medicine, South Kensington, London, United Kingdom. f.culley@ic.ac.uk
authors.forename: F J
authors.initials: FJ
authors.lastname: Culley
authors.forename: A
authors.initials: A
authors.lastname: Brown
authors.forename: D M
authors.initials: DM
authors.lastname: Conroy
authors.forename: I
authors.initials: I
authors.lastname: Sabroe
authors.forename: D I
authors.initials: DI
authors.lastname: Pritchard
authors.forename: T J
authors.initials: TJ
authors.lastname: Williams
mesh_terms: D000818:Animals; D000937:Antigen-Antibody Reactions; D001681:Biological Assay; D002464:Cell Migration Inhibition; D054413:Chemokine CCL11; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil / Q000008:administration & dosage / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism*; D016207:Cytokines / Q000008:administration & dosage / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D004804:Eosinophils / Q000276:immunology; D006168:Guinea Pigs; D015801:Helminth Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D007271:Injections, Intradermal; D007700:Kinetics; D007975:Leukotriene B4 / Q000008:administration & dosage; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D017221:Necator americanus / Q000201:enzymology* / Q000276:immunology*; D010436:Pepstatins / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity / Q000276:immunology
chemical_list: C515456:CCL11 protein, human; D054413:Chemokine CCL11; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil; D016207:Cytokines; D015801:Helminth Proteins; D010436:Pepstatins; D010618:Phenanthrolines; D011480:Protease Inhibitors; C014195:Streptomyces pepsin inhibitor; D007975:Leukotriene B4; D008666:Metalloendopeptidases; C031375:pepstatin