Journal Article

Cell Entry and Trafficking of Human Adenovirus Bound to Blood Factor X Is Determined by the Fiber Serotype and Not Hexon: Heparan Sulfate Interaction

Lindholm, Leif, Gonzalez, Gaëlle, Fender, Pascal, French Cystic Fibrosis Foundation (Vaincre la Mucoviscidose, VLM) TG0801 TG0901, Henning, Petra, Unit of Virus Host Cell Interactions = Biologie structurale des interactions entre virus et cellule hôte (UVHCI), Grichine, Alexei, Rétrovirus et Pathologie Comparée, Contrat d'Interface HCL-INSERM, Region Rhone-Alpes, ANRS, Got-A-Gene AB, Hong, Saw-See, Boulanger, Pierre, Corjon, Stephanie

2011

Overview

Human adenovirus serotype 5 (HAdV5)-based vectors administered intravenously accumulate in the liver as the result of their direct binding to blood coagulation factor X (FX) and subsequent interaction of the FX-HAdV5 complex with heparan sulfate proteoglycan (HSPG) at the surface of liver cells. Intriguingly, the serotype 35 fiber-pseudotyped vector HAdV5F35 has liver transduction efficiencies 4-logs lower than HAdV5, even though both vectors carry the same hexon capsomeres. In order to reconcile this apparent paradox, we investigated the possible role of other viral capsid proteins on the FX/HSPG-mediated cellular uptake of HAdV5-based vectors. Using CAR-and CD46-negative CHO cells varying in HSPG expression, we confirmed that FX bound to serotype 5 hexon protein and to HAdV5 and HAdV5F35 virions via its Gla-domain, and enhanced the binding of both vectors to surface-immobilized hypersulfated heparin and cellular HSPG. Using penton mutants, we found that the positive effect of FX on HAdV5 binding to HSPG and cell transduction did not depend on the penton base RGD and fiber shaft KKTK motifs. However, we found that FX had no enhancing effect on the HAdV5F35-mediated cell transduction, but a negative effect which did not involve the cell attachment or endocytic step, but the intracellular trafficking and nuclear import of the FX-HAdV5F35 complex. By cellular imaging, HAdV5F35 particles were observed to accumulate in the late endosomal compartment, and were released in significant amounts into the extracellular medium via exocytosis. We showed that the stability of serotype 5 hexon: FX interaction was higher at low pH compared to neutral pH, which could account for the retention of FX-HAdV5F35 complexes in the late endosomes. Our results suggested that, despite the high affinity interaction of hexon capsomeres to FX and cell surface HSPG, the adenoviral fiber acted as the dominant determinant of the internalization and trafficking pathway of HAdV5-based vectors.

• Publisher: CCSD,Public Library of Science,Public Library of Science (PLoS)
• Subject: Acids / Adenoviruses / Adenoviruses, Human - classification / Adenoviruses, Human - metabolism / Amino Acid Motifs / Animals / Antigens / Binding / Biology / Blood / Blood coagulation / Capsid Proteins - metabolism / CD46 antigen / Cell Adhesion / Cell Compartmentation / Cell Membrane - metabolism / Cell surface / CHO Cells / Clinical Medicine / Coagulation / Coagulation factor X / Coagulation factors / Cricetinae / Cricetulus / Cytotoxicity / Dendritic cells / Endocytosis / Endosomes / Exocytosis / Expression vectors / Factor X - chemistry / Factor X - metabolism / Green Fluorescent Proteins - metabolism / HEK293 Cells / Heparan sulfate / Heparan sulfate proteoglycans / Heparin / Heparitin Sulfate - metabolism / Hepatocytes / Humans / Internalization / Intracellular Space - metabolism / Klinisk medicin / Life Sciences / Liver / Mutants / Mutation - genetics / Nuclear transport / Oligopeptides - metabolism / Pathology / pH effects / Protein Binding / Protein Structure, Tertiary / Proteins / Serotyping / Sulfate / Sulfates / Surface Plasmon Resonance / Transduction, Genetic / Viral proteins / Virion - metabolism / Virion - ultrastructure / Virions / Virus Internalization / Viruses
• ISBN: 0002910522000