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Journal of Virology, August 2003, p. 8263-8271, Vol. 77, No. 15
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.15.8263-8271.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Replication-Deficient Human Adenovirus Type 35 Vectors for Gene Transfer and Vaccination: Efficient Human Cell Infection and Bypass of Preexisting Adenovirus Immunity

Ronald Vogels,1 David Zuijdgeest,1 Richard van Rijnsoever,1 Eric Hartkoorn,1 Irma Damen,1 Marie-Pierre de Béthune,2 Stefan Kostense,1 Germaine Penders,1 Niels Helmus,1 Wouter Koudstaal,1 Marco Cecchini,3 Antoinette Wetterwald,3 Mieke Sprangers,1 Angelique Lemckert,1 Olga Ophorst,1 Björn Koel,1 Michelle van Meerendonk,1 Paul Quax,4 Laura Panitti,2 Jos Grimbergen,4 Abraham Bout,1 Jaap Goudsmit,1 and Menzo Havenga1*

Vaccine R&D Division, Crucell Holland BV, 2301CA Leiden,1 Division of Vascular and Connective Tissue Research, Gaubius Laboratory, 2301CE Leiden, The Netherlands,4 Tibotec-Virco NV, 2800 Mechelen, Belgium Urology,2 Clinical Research, University of Bern, C-813 Bern, Switzerland3

Received 17 January 2003/ Accepted 30 April 2003

Replication-deficient human adenovirus type 5 (Ad5) can be produced to high titers in complementing cell lines, such as PER.C6, and is widely used as a vaccine and gene therapy vector. However, preexisting immunity against Ad5 hampers consistency of gene transfer, immunological responses, and vector-mediated toxicities. We report the identification of human Ad35 as a virus with low global prevalence and the generation of an Ad35 vector plasmid system for easy insertion of heterologous genes. In addition, we have identified the minimal sequence of the Ad35-E1B region (molecular weight, 55,000 [55K]), pivotal for complementation of fully E1-lacking Ad35 vector on PER.C6 cells. After stable insertion of the 55K sequence into PER.C6 cells a cell line was obtained (PER.C6/55K) that efficiently transcomplements both Ad5 and Ad35 vectors. We further demonstrate that transduction with Ad35 is not hampered by preexisting Ad5 immunity and that Ad35 efficiently infects dendritic cells, smooth muscle cells, and synoviocytes, in contrast to Ad5.

* Corresponding author. Mailing address: Vaccine R&D Division, Crucell Holland BV, P.O. Box 2048, 2301CA Leiden, The Netherlands. Phone: 31 (0) 71 5248736. Fax: 31 (0) 71 5248702. E-mail: m.havenga{at}crucell.com.

Journal of Virology, August 2003, p. 8263-8271, Vol. 77, No. 15
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.15.8263-8271.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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