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A more recent version of this article appeared on September 26, 2003
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M303496200v1
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Papers In Press, published online ahead of print July 10, 2003
J. Biol. Chem, 10.1074/jbc.M303496200
Submitted on April 4, 2003
Revised on July 8, 2003
Accepted on July 10, 2003

The maturation of murine dendritic cells induced by human Adenovirus is mediated by the fiber knob domain

Valérie Molinier-Frenkel, Armelle Prévost-Blondel, Saw-See Hong, Renée Lengagne, Sarah Boudaly, Maria K. Magnusson, Pierre Boulanger, and Jean-Gérard Guillet

Department of Virology, RTH Laennec, Lyon 69372

Corresponding Author: Pierre.Boulanger{at}laennec.univ-lyon1.fr

We investigated the mechanism of adenovirus serotype 5 (Ad5)-mediated maturation of bone marrow-derived murine dendritic cells (DC) using (i) Ad5 vectors with wild-type capsid (AdE1°, AdGFP), (ii) Ad5 vector mutant deleted of the fiber C-terminal knob domain (AdGFPdeltaknob), and (iii) capsid components isolated from Ad5-infected cells or expressed as recombinant proteins, hexon, penton, penton base, full-length fiber, fiber knob, and fiber mutants. We found that penton capsomer (penton-base linked to its fiber projection), full-length fiber protein and its isolated knob domain, were all capable of inducing DC maturation, whereas no significant DC maturation was observed for hexon or penton-base alone. This capacity was severely reduced for AdGFPdeltaknob, and for fiber protein deletion mutants lacking the beta-stranded region F of the knob (residues L485-T486). The DC maturation effect was fully retained in a recombinant fiber protein deleted of the HI loop (FideltaHI), a fiber deletion mutant which failed to trimerize, suggesting that the fiber knob-mediated DC activation did not depend on the integrity of the HI loop and on the trimeric status of the fiber. Interestingly, peptide-pulsed DC that had been stimulated with Ad5 knob protein induced a potent CD8+ T cell response in vivo.


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