title: Membrane-bound CC chemokine inhibitor 35K provides localized inhibition of CC chemokine activity in vitro and in vivo.
abstract: CC chemokines mediate mononuclear cell recruitment and activation in chronic inflammation. We have shown previously that gene transfer using recombinant adenoviruses, encoding a soluble CC chemokine-binding protein of vaccinia virus 35K, can dramatically reduce atherosclerosis and vein graft remodeling in apolipoprotein E knockout mice. In this study, we report the development of a membrane-bound form of 35K (m35K), tagged with GFP, which allows for localized, broad-spectrum CC chemokine blockade. In vitro experiments indicate that m35K-expressing cells no longer undergo CC chemokine-induced chemotaxis, and m35K-expressing cells can locally deplete the CC chemokines RANTES (CCL5) and MIP-1alpha (CCL3) from supernatant medium. This sequestration of CC chemokines can prevent chemotaxis of bystander cells to CC, but not CX(3)C chemokines. Intraperitoneal injection of mice with an adenovirus-encoding m35K leads to a significant (44%) decrease in leukocyte recruitment into the peritoneal cavity in a sterile peritonitis model. Intravenous adenovirus-encoding m35K delivery leads to m35K expression in hepatocytes, which confers significant protection against liver damage (75% reduction in liver enzymes) in a Con A-induced hepatitis model. In summary, we have generated a membrane-bound CC chemokine-binding protein (m35K) that provides localized broad-spectrum CC chemokine inhibition in vitro and in vivo. m35K may be a useful tool to study the role of CC chemokines in leukocyte trafficking and block the recruitment of monocytes in chronic inflammation.
authors.affiliation: Department of Cardiovascular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom.
authors.forename: Christina A
authors.initials: CA
authors.lastname: Bursill
authors.forename: Jenna L
authors.initials: JL
authors.lastname: Cash
authors.forename: Keith M
authors.initials: KM
authors.lastname: Channon
authors.forename: David R
authors.initials: DR
authors.lastname: Greaves
mesh_terms: D000818:Animals; D019742:Chemokines, CC / Q000819:agonists*; D002633:Chemotaxis / Q000187:drug effects; D004195:Disease Models, Animal; D049452:Green Fluorescent Proteins / Q000235:genetics; D006505:Hepatitis / Q000473:pathology / Q000628:therapy; D008565:Membrane Proteins; D051379:Mice; D010538:Peritonitis / Q000628:therapy; D014616:Vaccinia virus / Q000737:chemistry; D014759:Viral Envelope Proteins / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*
chemical_list: D019742:Chemokines, CC; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; C072495:immunodominant protein p35, vaccinia virus; D049452:Green Fluorescent Proteins