1. ¹College of Pharmacy, Division of Pharmaceutics, The University of Texas at Austin, Austin, TX, USA
2. ²The Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA
3. ³Dipartimento di Biochimica e Biotecnologie Mediche, University of Naples 'Federico II'
4. ⁴CEINGE-Biotecnologie Avanzate, Napoli, Italy
5. ⁵Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA

Correspondence: Dr MA Croyle, College of Pharmacy, Division of Pharmaceutics, The University of Texas at Austin, PHR 4.214D, 1 University Station #A1920, 2409 W. University Avenue, Austin, TX 78712, USA

Received 22 July 2004; Accepted 4 October 2004; Published online 13 January 2005.

Abstract

Transgene expression from helper-dependent adenoviral (HD-Ad) vectors is effective and long lasting, but not permanent. Their use is also limited by the host response against capsid proteins that precludes successful gene expression upon readministration. In this report, we test the hypothesis that PEGylation of HD-Ad reduces its toxicity and promotes transgene expression upon readministration. PEGylation did not compromise transduction efficiency in vitro and in vivo and reduced peak serum IL-6 levels two-fold. IL-12 and TNF- levels were reduced three- and seven-fold, respectively. Thrombocytopenia was not detected in mice treated with the PEGylated vector. Serum transaminases were not significantly elevated in mice treated with either vector. Mice immunized with 1 10¹¹ particles of unmodified HD-Ad expressing human alpha-1 antitrypsin (hA1AT) were rechallenged 28 days later with 8 10¹⁰ particles of unmodified or PEG-conjugated vector expressing beta-galactosidase. Trace levels of beta-galactosidase (52.2319.2 pg/mg protein) were detected in liver homogenates of mice that received two doses of unmodified HD-Ad. Mice rechallenged with PEGylated HD-Ad produced significant levels of beta-galactosidase (5.10.4 10⁵ pg/mg protein, P=0.0001). This suggests that PEGylation of HD-Ad vectors may be appropriate for their safe and efficient use in the clinic.