title: E-selectin is a viable route of infection for polymer-coated adenovirus retargeting in TNF-α-activated human umbilical vein endothelial cells.
abstract: BACKGROUND
  E-selectin is an attractive endothelial cell surface marker in inflammation and cancer.
  
  
  OBJECTIVE
  We sought to investigate retargeting of adenovirus via E-selectin as a viable pathway of infection in tumor necrosis factor-α (TNF-α)-activated human umbilical vein endothelial cells (HUVECs).
  
  
  METHODS
  E1, E3-deleted Ad5 expressing cytomegalovirus immediate-early (CMV IE) promoter-driven luciferase (Adluc) was coated with an amino-reactive multivalent hydrophilic polymer based on poly [N-(2-hydroxypropyl) methacrylamide] to generate pHPMA-adenovirus (pcAdluc). This was then retargeted by covalent attachment of a mouse antihuman E-selectin monoclonal antibody (MHES mAb), purified from the H18/7 hybridoma cell line (MHESpcAdluc).
  
  
  RESULTS
  MHESpcAdluc was efficiently taken up into HUVECs, generating a high level of transduction in TNF-α-treated E-selectin positive cells but not in untreated receptor-negative cells. Specific retargeting of MHESpcAdluc was demonstrated through reduced transduction of stimulated HUVEC when incubated in the presence of free E-selectin antibodies.
  
  
  CONCLUSIONS
  Our results suggest that E-selectin could be a valuable target for gene transfer strategies internalizing polymer-coated modified adenovirus particles through a viable receptor-mediated endocytosis pathway, generating adequate levels of transgene expression per virus genome copy without compromising the specific activity of the parental virus.
authors.affiliation: Department of Clinical Pharmacology, University of Oxford, Oxford, Oxfordshire, UK.
authors.forename: Houria
authors.initials: H
authors.lastname: Bachtarzi
authors.forename: Mark
authors.initials: M
authors.lastname: Stevenson
authors.forename: Vladimir
authors.initials: V
authors.lastname: Subr
authors.forename: Leonard W
authors.initials: LW
authors.lastname: Seymour
authors.forename: Kerry D
authors.initials: KD
authors.lastname: Fisher
mesh_terms: D000178:Acrylamides / Q000737:chemistry*; D000256:Adenoviridae / Q000235:genetics*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D019040:E-Selectin / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D004705:Endocytosis; D042783:Endothelial Cells / Q000276:immunology / Q000378:metabolism / Q000821:virology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000821:virology; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism; D008024:Ligands; D008156:Luciferases / Q000235:genetics; D016133:Polymerase Chain Reaction; D011991:Receptors, Virus / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D000178:Acrylamides; D000911:Antibodies, Monoclonal; D019040:E-Selectin; D007074:Immunoglobulin G; D008024:Ligands; D011991:Receptors, Virus; C546936:SELE protein, human; D014409:Tumor Necrosis Factor-alpha; C085301:adenovirus receptor; D008156:Luciferases; C032976:N-(2-hydroxypropyl)methacrylamide