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Journal of Virology, November 1999, p. 9137-9144, Vol. 73, No. 11
Virology Group, Veterinary Infectious Disease
Organization, University of Saskatchewan, Saskatoon, Saskatchewan,
Canada S7N 5E3,1 and Gene Therapy
Department, Transgene, S.A., 67000 Strasbourg,
France2
Received 30 March 1999/Accepted 26 July 1999
Although recombinant human adenovirus (HAV)-based vectors offer
several advantages for somatic gene therapy and vaccination over other
viral vectors, it would be desirable to develop alternative vectors
with prolonged expression and decreased toxicity. Toward this
objective, a replication-defective bovine adenovirus type 3 (BAV-3) was
developed as an expression vector. Bovine cell lines designated VIDO R2
(HAV-5 E1A/B-transformed fetal bovine retina cell [FBRC] line) and
6.93.9 (Madin-Darby bovine kidney [MDBK] cell line expressing E1
proteins) were developed and found to complement the E1A deletion in
BAV-3. Replication-defective BAV-3 with a 1.7-kb deletion removing most
of the E1A and E3 regions was constructed. This virus could be grown in
VIDO R2 or 6.93.9 cells but not in FBRC or MDBK cells. The results
demonstrated that the E1 region of HAV-5 has the capacity to transform
bovine retina cells and that the E1A region of HAV-5 can complement
that of BAV-3. A replication-defective BAV-3 vector expressing bovine herpesvirus type 1 glycoprotein D from the E1A region was made. A
similar replication-defective vector expressing the
hemagglutinin-esterase gene of bovine coronavirus from the E3 region
was isolated. Although these viruses grew less efficiently than the
replication-competent recombinant BAV-3 (E3 deleted), they are suitable
for detailed studies with animals to evaluate the safety, duration of
foreign gene expression, and ability to induce immune responses. In
addition, a replication-competent recombinant BAV-3 expressing green
fluorescent protein was constructed and used to evaluate the host range
of BAV-3 under cell culture conditions. The development of bovine E1A-complementing cell lines and the generation of
replication-defective BAV-3 vectors is a major technical advancement
for defining the use of BAV-3 as vector for vaccination against
diseases of cattle and somatic gene therapy in humans.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Replication-Defective Bovine Adenovirus Type 3 as
an Expression Vector
*
Corresponding author. Mailing address: VIDO, 120 Veterinary Rd., University of Saskatchewan, Saskatoon, Saskatchewan,
Canada S7N 5E3. Phone: (306) 966-7482. Fax: (306) 966-7478. E-mail:
tikoo{at}sask.usask.ca.
Published with the permission of the Director of VIDO as journal
series no. 258.
Present address: Genetic Therapy, Inc., Gaithersburg, MD 20878.
Journal of Virology, November 1999, p. 9137-9144, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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