Replication-deficient human adenovirus type 35 vectors for gene transfer and vaccination: efficient human cell infection and bypass of preexisting adenovirus immunity.
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.
Vogels
Ronald
R
Vaccine R&D Division, Crucell Holland BV, 2301CA Leiden, The Netherlands.
Zuijdgeest
David
D
van Rijnsoever
Richard
R
Hartkoorn
Eric
E
Damen
Irma
I
de Béthune
Marie-Pierre
MP
Kostense
Stefan
S
Penders
Germaine
G
Helmus
Niels
N
Koudstaal
Wouter
W
Cecchini
Marco
M
Wetterwald
Antoinette
A
Sprangers
Mieke
M
Lemckert
Angelique
A
Ophorst
Olga
O
Koel
Björn
B
van Meerendonk
Michelle
M
Quax
Paul
P
Panitti
Laura
L
Grimbergen
Jos
J
Bout
Abraham
A
Goudsmit
Jaap
J
Havenga
Menzo
M
D017501:Adenovirus E1B Proteins / Q000737:chemistry / Q000235:genetics; D000260:Adenoviruses, Human / Q000235:genetics / Q000276:immunology* / Q000502:physiology*; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000821:virology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000276:immunology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009130:Muscle, Smooth / Q000166:cytology / Q000821:virology; D009500:Neutralization Tests; D010957:Plasmids; D013583:Synovial Membrane / Q000166:cytology / Q000821:virology; D014611:Vaccination; D019065:Virus Assembly; D014779:Virus Replication
D017501:Adenovirus E1B Proteins; D000914:Antibodies, Viral