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J. Biol. Chem., Vol. 278, Issue 39, 37175-37182, September 26, 2003

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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 

From the Département d'Immunologie, Institut Cochin, INSERM U567, CNRS UMR 8104, Laboratoire membre de l'Institut Fédératif de Recherche 116, Université R. Descartes, 27 rue du Faubourg Saint Jacques, 75014 Paris, France, the ^**Laboratoire de Virologie et Pathogénèse Virale, CNRS UMR 5537, Faculté de Médecine R.T.H. Laennec, 7, Rue Guillaume Paradin, 69372 Lyon Cedex 08, France, Laboratoire de Pathologie Métabolique et Hormonale du Développement, INSERM U342, Hôpital Saint Vincent de Paul, 75014 Paris, France, and the Got-A-Gene AB, Stena Center 1B, SE 41292 Göteborg, Sweden

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 (AdGFPknob); 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 AdGFPknob and for fiber protein deletion mutants lacking the -stranded region F of the knob (residues Leu-485–Thr-486). The DC maturation effect was fully retained in a recombinant fiber protein deleted of the HI loop (FiHI), a fiber (Fi) deletion mutant that 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.

Received for publication, April 4, 2003 , and in revised form, July 8, 2003.

^* This study was supported by grants from the French Cystic Fibrosis Association "Vaincre la Mucoviscidose" (VLM) and from the Association Française contre les Myopathies (AFM). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

^|| Supported by a fellowship from the Association Française de Recherche contre le Cancer.

^¶ Supported by a fellowship from VLM. To whom correspondence may be addressed: Laboratoire d'Immunologie, Hôpital Henri Mondor, 51, av du Maréchal de Lattre de Tassigny, 94010 Créteil Cedex, France. Tel.: 33-1-4981-2298; Fax: 33-1-4981-2897; E-mail: valerie.frenkel{at}hmn.ap-hop-paris.fr.

^¶¶ To whom correspondence may be addressed: Tel.: 33-4-7877-8621; Fax: 33-4-7877-8751; E-mail: Pierre.Boulanger{at}laennec.univ-lyon1.fr.

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