Affiliation:
1. Max-von-Pettenkofer Institute, Gene
Center, Ludwig-Maximilians-University, 81377 Munich,
Germany
2. University
of Warwick, Department of Biological Sciences, Coventry CV4 7AL, United
Kingdom
Abstract
ABSTRACT
Until
recently, adenovirus (Ad)-mediated gene therapy was almost exclusively
based on human Ad type 5 (Ad5). Preexisting immunity and the limited,
coxsackievirus and adenovirus receptor-dependent tropism of Ad5
stimulated attempts to exploit the natural diversity in tropism of the
other 50 known human Ad serotypes. Aiming in particular at
immunotherapy and vaccination, we have screened representative
serotypes from different Ad species for their ability to infect
dendritic cells. Ad19a, an Ad from species D, was selected for
development as a new vector for vaccination and cancer gene therapy. To
clone and manipulate its genome, we have developed a novel methodology,
coined “exposon mutagenesis,” that allows the rapid and
precise introduction of virtually any genetic alteration (deletions,
point mutations, or insertions) into recombinant Ad bacterial
artificial chromosomes. The versatility of the system was exemplified
by deleting the E3 region of Ad19a, by specifically knocking out
expression of a species-specific E3 gene, E3/49K, and by reinserting
E3/49K into an E3 null Ad19a mutant. The technology requires only
limited sequence information and is applicable to other Ad species.
Therefore, it should be extremely valuable for the analysis of gene
functions from any Ad species. In addition, a basic,
replication-defective E1- and E3-deleted Ad19a vector expressing GFP
(Ad19aGFP) was generated. This new vector based on species D Ads
exhibits a very promising tropism for lymphoid and muscle cells and
shows great potential as an alternative vector for transduction of cell
types that are resistant to or only poorly transduced by conventional
Ad5-based
vectors.
Publisher
American Society for Microbiology
Subject
Virology,Insect Science,Immunology,Microbiology
Cited by
35 articles.
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