A Partial E3 Deletion in Replication-Defective Adenoviral Vectors Allows for Stable Expression of Potentially Toxic Transgene Products.
Adenovirus (Ad) is used extensively for construction of viral vectors, most commonly with deletion in its E1 and/or E3 genomic regions. Previously, our attempts to insert envelope proteins (Env) of HIV-1 into such vectors based on chimpanzee-derived Ad (AdC) viruses were thwarted. Here, we describe that genetic instability of an E1- and E3-deleted AdC vector of serotype C6 expressing Env of HIV-1 can be overcome by reinsertion of E3 sequences with anti-apoptotic activities. This partial E3 deletion presumably delays premature death of HEK-293 packaging cell lines due to Env-induced cell apoptosis. The same partial E3 deletion also allows for the generation of stable glycoprotein 140 (gp140)- and gp160-expressing Ad vectors based on AdC7, a distinct AdC serotype. Env-expressing AdC vectors containing the partial E3 deletion are genetically stable upon serial cell culture passaging, produce yields comparable to those of other AdC vectors, and induce transgene product-specific antibody responses in mice. A partial E3 deletion thereby allows expansion of the repertoire of transgenes that can be expressed by Ad vectors.
1 The Wistar Institute , Philadelphia, Pennsylvania.
Larissa H
LH
Haut
1 The Wistar Institute , Philadelphia, Pennsylvania.|2 Current address: Clinical Molecular Regulation Section/Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health , Bethesda, Maryland.
Amanda L
AL
Gill
1 The Wistar Institute , Philadelphia, Pennsylvania.
Raj K
RK
Kurupati
1 The Wistar Institute , Philadelphia, Pennsylvania.
Ang
A
Bian
1 The Wistar Institute , Philadelphia, Pennsylvania.
Yan
Y
Li
1 The Wistar Institute , Philadelphia, Pennsylvania.
Wynetta
W
Giles-Davis
1 The Wistar Institute , Philadelphia, Pennsylvania.
Zhiquan
Z
Xiang
1 The Wistar Institute , Philadelphia, Pennsylvania.
Xiang Yang
XY
Zhou
1 The Wistar Institute , Philadelphia, Pennsylvania.
Hildegund C J
HC
Ertl
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics* / Q000627:therapeutic use; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D051379:Mice; D065288:Serogroup; D019076:Transgenes / Q000235:genetics; D014779:Virus Replication / Q000235:genetics; D054299:env Gene Products, Human Immunodeficiency Virus / Q000096:biosynthesis* / Q000627:therapeutic use
D054299:env Gene Products, Human Immunodeficiency Virus
HIV-1; genetic stability; immune responses; vaccine