The contamination of adenovirus (Ad) stocks with adeno-associated viruses (AAV) is usually unnoticed, and it has been associated with lower Ad yields upon large-scale production. During Ad propagation, AAV contamination needs to be detected routinely by polymerase chain reaction without symptomatic suspicion. In this study, we describe that the coinfection of either Ad wild type 5 or oncolytic Ad with AAV results in a large-plaque phenotype associated with an accelerated release of Ad from coinfected cells. This accelerated release was accompanied with the expected decrease in Ad yields in two out of three cell lines tested. Despite this lower Ad yield, coinfection with AAV accelerated cell death and enhanced the cytotoxicity mediated by Ad propagation. Intratumoral coinjection of Ad and AAV in two xenograft tumor models improved antitumor activity and mouse survival. Therefore, we conclude that accidental or intentional AAV coinfection has important implications for Ad-mediated virotherapy.
Get full access to this article
View all access options for this article.
References
1.
AgrawalN., et al. (2002). Temporal acceleration of the human papillomavirus life cycle by adeno-associated virus (AAV) type 2 superinfection in natural host tissue. Virology, 297, 203–210.
2.
AlemanyR. (2007). Cancer selective adenoviruses. Mol. Aspects Med., 28, 42–58.
3.
CarterB.J., et al. (1979). Adeno-associated virus autointerference. Virology, 92, 449–462.
4.
CascalloM., et al. (2007). Systemic toxicity-efficacy profile of ICOVIR-5, a potent and selective oncolytic adenovirus based on the pRB pathway. Mol. Ther., 15, 1607–1615.
5.
CasperJ.M., et al. (2005). Identification of an adeno-associated virus Rep protein binding site in the adenovirus E2a promoter. J. Virol., 79, 28–38.
6.
CoukosG., et al. (1999). Use of carrier cells to deliver a replication-selective herpes simplex virus-1 mutant for the intraperitoneal therapy of epithelial ovarian cancer. Clin. Cancer Res., 5, 1523–1537.
7.
DoroninK., et al. (2003). Overexpression of the ADP (E3-11.6K) protein increases cell lysis and spread of adenovirus. Virology, 305, 378–387.
8.
GaoG., et al. (2004). Clades of adeno-associated viruses are widely disseminated in human tissues. J. Virol., 78, 6381–6388.
9.
Garcia-CastroJ., et al. (2005). Tumor cells as cellular vehicles to deliver gene therapies to metastatic tumors. Cancer Gene Ther., 12, 341–349.
10.
GrimmD., et al. (2003). Preclinical in vivo evaluation of pseudotyped adeno-associated virus vectors for liver gene therapy. Blood, 102, 2412–2419.
11.
GrimmD., et al. (2008). In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J. Virol., 82, 5887–5911.
12.
GrosA. (2010). Adenovirus release from the infected cell as a key factor for adenovirus oncolysis. Open Gene Ther. J., 3, 24–30.
13.
GrosA., et al. (2008). Bioselection of a gain of function mutation that enhances adenovirus 5 release and improves its antitumoral potency. Cancer Res., 68, 8928–8937.
14.
HamadaK., et al. (2007). Carrier cell-mediated delivery of a replication-competent adenovirus for cancer gene therapy. Mol. Ther., 15, 1121–1128.
15.
HogganM.D., et al. (1966). Studies of small DNA viruses found in various adenovirus preparations: physical, biological, and immunological characteristics. Proc. Natl. Acad. Sci. USA, 55, 1467–1474.
16.
JingX.J., et al. (2001). Inhibition of adenovirus cytotoxicity, replication, and E2a gene expression by adeno-associated virus. Virology, 291, 140–151.
17.
KhleifS.N., et al. (1991). Inhibition of cellular transformation by the adeno-associated virus rep gene. Virology, 181, 738–741.
18.
KwonO.J., et al. (2011). Viral genome DNA/lipoplexes elicit in situ oncolytic viral replication and potent antitumor efficacy via systemic delivery. J. Control Release, 155, 317–325.
19.
MullerO.J., et al. (2006). Improved cardiac gene transfer by transcriptional and transductional targeting of adeno-associated viral vectors. Cardiovasc. Res., 70, 70–78.
20.
Puig-SausC., et al. (2012). Adenovirus i-leader truncation bioselected against cancer-associated fibroblasts to overcome tumor stromal barriers. Mol. Ther., 20, 54–62.
21.
RamachandraM., et al. (2001). Re-engineering adenovirus regulatory pathways to enhance oncolytic specificity and efficacy. Nat. Biotechnol., 19, 1035–1041.
22.
RojasJ.J., et al. (2010). Minimal RB-responsive E1A promoter modification to attain potency, selectivity, and transgene-arming capacity in oncolytic adenoviruses. Mol. Ther., 18, 1960–1971.
23.
RussellS.J., et al. (2012). Oncolytic virotherapy. Nat. Biotechnol., 30, 658–670.
24.
SauthoffH., et al. (2000). Deletion of the adenoviral E1b-19kD gene enhances tumor cell killing of a replicating adenoviral vector. Hum. Gene Ther., 11, 379–388.
25.
SauthoffH., et al. (2002). Late expression of p53 from a replicating adenovirus improves tumor cell killing and is more tumor cell specific than expression of the adenoviral death protein. Hum. Gene Ther., 13, 1859–1871.
26.
SchmidtM., et al. (2000). Adeno-associated virus type 2 Rep78 induces apoptosis through caspase activation independently of p53. J. Virol., 74, 9441–9450.
27.
SchmidtM., et al. (2006). Identification and characterization of novel adeno-associated virus isolates in ATCC virus stocks. J. Virol., 80, 5082–5085.
28.
StantonR.J., et al. (2008). Re-engineering adenovirus vector systems to enable high-throughput analyses of gene function. Biotechniques, 45, 659–662, 664–658.
29.
SubramanianT., et al. (2006). Genetic identification of adenovirus type 5 genes that influence viral spread. J. Virol., 80, 2000–2012.
30.
TimpeJ.M., et al. (2006). Effects of adeno-associated virus on adenovirus replication and gene expression during coinfection. J. Virol., 80, 7807–7815.
31.
TimpeJ.M., et al. (2007). Adeno-associated virus induces apoptosis during coinfection with adenovirus. Virology, 358, 391–401.
32.
TollefsonA.E., et al. (1996a). The E3-11.6-kDa adenovirus death protein (ADP) is required for efficient cell death: characterization of cells infected with adp mutants. Virology, 220, 152–162.
33.
TollefsonA.E., et al. (1996b). The adenovirus death protein (E3-11.6K) is required at very late stages of infection for efficient cell lysis and release of adenovirus from infected cells. J. Virol., 70, 2296–2306.
34.
VillanuevaA., et al. (1998). Disruption of the antiproliferative TGF-beta signaling pathways in human pancreatic cancer cells. Oncogene, 17, 1969–1978.
35.
WeinL.M., et al. (2003). Validation and analysis of a mathematical model of a replication-competent oncolytic virus for cancer treatment: implications for virus design and delivery. Cancer Res., 63, 1317–1324.
36.
WillmonC., et al. (2009). Cell carriers for oncolytic viruses: Fed Ex for cancer therapy. Mol. Ther., 17, 1667–1676.
37.
ZhouC., and TrempeJ.P. (1999). Induction of apoptosis by cadmium and the adeno-associated virus Rep proteins. Virology, 261, 280–287.
38.
ZhouC., et al. (1999). Enhancement of UV-induced cytotoxicity by the adeno-associated virus replication proteins. Biochim. Biophys. Acta, 1444, 371–383.
39.
ZolotukhinS., et al. (1999). Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Ther., 6, 973–985.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
