E-selectin is a viable route of infection for polymer-coated adenovirus retargeting in TNF-α-activated human umbilical vein endothelial cells.

Journal article

BACKGROUND: E-selectin is an attractive endothelial cell surface marker in inflammation and cancer. PURPOSE: 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. DISCUSSION AND CONCLUSION: 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: Bachtarzi, H; Stevenson, M; Subr, V; Seymour, L; Fisher, K

• Publication status: Published
• Journal: Journal of drug targeting
• Volume: 19
• Issue: 8
• Pages: 690-700
• Publication date: 2011-09-01
• DOI: 10.3109/1061186x.2010.547585
• EISSN: 1029-2330
• ISSN: 1061-186X
• Language: English
• Keywords: Endothelium, Vascular; Polymerase Chain Reaction; Endocytosis; Immunoglobulin G; Acrylamides; Transduction, Genetic; Gene Transfer Techniques; Genetic Vectors; Flow Cytometry; Tumor Necrosis Factor-alpha; Ligands; Endothelial Cells; Luciferases; Human Umbilical Vein Endothelial Cells; E-Selectin; Receptors, Virus; Adenoviridae; Antibodies, Monoclonal; Transgenes; Humans