Membrane-bound CC chemokine inhibitor 35K provides localized inhibition of CC chemokine activity in vitro and in vivo.
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.
Bursill
Christina A
CA
Department of Cardiovascular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom.
Cash
Jenna L
JL
Channon
Keith M
KM
Greaves
David R
DR
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*
D019742:Chemokines, CC; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; C072495:immunodominant protein p35, vaccinia virus; D049452:Green Fluorescent Proteins