Intracellular trafficking of a fiber-modified adenovirus using lipid raft/caveolae endocytosis.
Most adenoviral vectors (HAdvs) elaborated for gene therapy are derived from serotype 5 viruses that use clathrin-coated vesicle endocytosis for cell entry. However, it appears that adenoviral vectors are able to take advantage of lipid raft/caveolae endocytosis to infect cells. In vivo targeting of a therapeutic gene to specific cells by vector engineering has become a major focus of gene therapy research. Yet, modification of adenoviral tropism, especially fiber gene engineering, can induce deficient intracellular trafficking of the viral particle, with a shift in subcellular localization resulting in extensive exocytosis. In this study we demonstrate that uptake of a fiber-modified adenovirus using lipid raft/caveolae endocytosis leads to non-altered intracellular trafficking without endosomal retention. Moreover, activation of lipid raft structures by this vector leads to the formation of "mega-caveosomes". These results demonstrate that, by forcing adenoviruses to take advantage of a non-clathrin, non-classical endocytic pathway, it is possible to compensate for the deficiency in endosomolysis that is associated with the use of some of the fiber-modified adenoviral constructs. Moreover, it renders such vectors ideal candidates for infecting human coxsackie and adenoviruses receptor (hCAR) negative cells.
Rogée
Sophie
S
Institut National de la Santé et de la Recherche Médicale U837, Lille, France.
Grellier
Elodie
E
Bernard
Claudine
C
Loyens
Anne
A
Beauvillain
Jean-Claude
JC
D'halluin
Jean-Claude
JC
Colin
Morvane
M
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D021941:Caveolae / Q000378:metabolism*; D002460:Cell Line; D002966:Clathrin / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D016253:Microscopy, Immunoelectron; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure
D002966:Clathrin