Adenoassociated viral aav vector with cmv promoter, sv40 polya signal, maximum capacity for insertion 2. The properties of constitutive promoters within adenoassociated viral aav vectors have not yet been fully characterized. The influence of sv40 polya on gene expression of baculovirus. Like other polyomaviruses, sv40 is a dna virus that has the potential to cause tumors in animals, but most often persists as a latent infection. The tumors investigated included ependymoma, all brain cancers combined, osteosarcoma, and mesothelioma. Sv40 promoter for highlevel constitutive expression of your gene of interest cmv promoter for highlevel constitutive expression of the cycle 3 gfpzeocin fusion sv40 origin for episomal replication and simple vector rescue in cell lines expressing the sv40 large t antigen the ptracercmv2. In the baculovirus prototype autographa californica multiple nucleopolyhedrovirus acmnpv, the polyhedrin promoter very late promoter transcribes its gene by a viral rna polymerase therefore there is no supporting. Other recently described viral agents include a bovine papilloma virus 12, and herpes simplex virus and, very recently, hiv 14. Intrathecal viral vector delivery of trastuzumab prevents or. The viral oncoproteins of dna tumor viruses ie, sv40 tantigen, hpv e6 protein, and adenovirus e1b55 kdae4 orf6 inactivate p53 via distinct mechanisms. In conclusion, sv40 intron is a potent strong intron element that increases transgene expression, which. Lentivirus production is influenced by sv40 large tantigen. We cloned the expression cassette nine copies of epo hre, sv40 minimal promoter, lacz gene, and sv40 polyadenylation signal into an aav vector. This page is informational only this vector is not available from addgene please contact the manufacturer for further details.
Introduction simian virus 40 sv40 was an inadvertent contaminant of the early poliovirus vaccines used throughout much of the world during the late 1950s and early 1960s. A new packaging cell line for sv40 vectors that eliminates. Simian virus 40 in humans infectious agents and cancer. Replication incompetent sv40 vectors in which the viral tantigen is replaced by the gene of interest are produced in vivo in cells that supply tantigen in trans, such as cos or cot cells, and. Hepatitis b surface antigen hbsag cdna was cloned downstream of two tandem sv40 early. Sv40 intron, a potent strong intron element that effectively. Dec 14, 2015 schematic diagram of viral constructs with and without sv40 polya signals for egfp gene expression under the acmnpv polh promoter control at the polh, egt and gp37 loci of acmnpv genome to test the functions of sv40 polya in the baculovirus expression vector system. Simian virus 40 sv40 is a promising viral vector for efficient gene transfer into a variety of human tissues, including the bone marrow and the liver 4, 5. Sv40 host range restriction and adenovirus helper factor the harvard community has made this article openly available. The vector of claim 4, wherein a vertebrate cell transfected therewith produces at least ca. Recombinant sv40 vectors rsv40 are good candidates for gene transfer, as they display some unique features.
Vaccine ingredients sv40 childrens hospital of philadelphia. Factors influencing the production of recombinant sv40. Sv40 was serendipitously discovered from a monkey cell line that was being used for the production of poliovirus. In conclusion, sv40 intron is a potent strong intron element that increases transgene expression, which can readily be used to more efficient transgenic protein production in cho cells. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. This may represent a pathway for sv40 infections in humans of unknown frequency, as there are reports of the detection and expression of sv40 tag and the presence of viral dna in cases of. Sv40 simian virus 40 viruses have several advantages for this purpose. Recently, the institute of medicine of the national academies concluded that the biological evidence is strong that sv40 is a transforming virus and that the biological evidence is of moderate. This may represent a pathway for sv40 infections in humans of unknown frequency, as there are reports of the detection and expression of sv40 tag and the presence of viral dna in cases of primary brain cancers in infants and young children 5, 71, 72, 117, 129, 3. Other viral vector systems that can be used for overexpression of proteins include vectors based on baculovirus, vaccinia virus, and herpes simplex virus. Testing of retroviral vectorbased human gene therapy. Viral dna isolated from rsvlucp3, rsvlucp4, rsvlucp2, and wtsv40 stocks was subjected to quantitative pcr using primers that amplified the sv40 late region vps or the luciferase transgene. We conclude that in vitro packaging is more flexible then previously realized, permitting packaging of at least 17 kb plasmid dna without the requirement for any viral sequences.
In this study, aav vectors, in which enhanced gfp expression was directed by one. Simian virus 40 sv40 is a monkey virus that was administered to human populations by contaminated vaccines which were produced in sv40 naturally infected monkey cells. More than 30 years of followup data were available. Emergent human pathogen simian virus 40 and its role in cancer.
Uses for the vectors include the expression of proteins in vitro and in vivo. A vector based on the sv40 origin of replication and chromosomal smars replicates episomally in cho cells article pdf available in nucleic acids research 272. During the past 4 years, polymerase chain reaction pcr assays have detected dna sequences related to sv40 an oncogenic simian polyomavirus in a variety of human tissues, especially choroid plexus tumors, ependymomas, mesotheliomas, and osteosarcomas 17. Identification of fam111a as an sv40 host range restriction. Alkek building for biomedical research request vectors through. Pdf a vector based on the sv40 origin of replication and. Adenoassociated viral vectormediated hypoxia response. Viral dna isolated from rsvlucp3, rsvlucp4, rsvlucp2, and wtsv40 stocks was subjected to quantitative pcr using primers that amplified the sv40 late region vps or the luciferase transgene luc. Sv40, we recently conducted a large, longterm retrospective followup study of birth cohorts exposed or unexposed to sv40contaminated poliovirus vaccine strickler et al. Sv40 infects primate cells, forcing its way inside and releasing its dna circle. The amplifications were done more than three times with each of six, four, and two different stocks of rsvlucp3, rsvlucp4, and rsvlucp2, respectively. The polyomavirus simian virus 40 sv40 is a potent dna tumor virus, and mounting evidence suggests that it is an emergent human pathogen 1, 10, 12, 39, 49, 50, 66, 111, 123. Wangshick ryu, in molecular virology of human pathogenic viruses, 2017. Intrathecal viral vector delivery of trastuzumab prevents or inhibits tumor growth of human her2positive xenografts in mice william t.
The sv40 host range mutant virus hr684 lacks the cterminal 24 residues of lt and has significantly reduced expression of early lt and late viral vp1. In this study, to determine whether rsv40 follows the same route as the wildtype. Recent molecular biology and epidemiological studies suggest that sv40 may be contagiously transmitted in humans by horizontal infection, independently from the earlier administration of sv40 contaminated vaccines. Simian virus 40 sv40 is an icosahedral nonenveloped polyomavirus with a.
Intrathecal viral vector delivery of trastuzumab prevents. We report here successful immunization using such an sv40 derived viral vector. Schematic illustration of the different dual vector approaches to expand the aav packaging capacity. Currently, lentiviral vectors for research and gene therapy are produced from 293t cells that are transiently transfected with plasmids encoding the vector and helper functions. Consequently, we devised a gene transfer system based on simian virus40 sv40 1 as a vector. They infect resting as well as cycling cells, they have a long term expression and they. Simian virus40 as a gene therapy vector mary ann liebert, inc. This is a free resource for the scientific community that is compiled by addgene. Thus, the mechanism of viral entry to deliver foreign genes is exploited, whereas the harmful effects of a productive viral infection in recipient cells are avoided. Propagation of these defective viruses required a helper virus and the recombinant viruses obtained could be grown only as a mixture. Sv40 t antigencontaining cells should not be considered more hazardous than the intact virus. Table i provides an overview of the testing required for cell banks, viral vector, and celltherapy product.
In particular, p53 is dysregulated via sequestration by sv40 tantigen, whereas p53 is dysregulated via degradation either by hpv e6associated ubiquitin e3 ligase or by adenovirus e1b55e4. Vaccine contamination with sv40 occurred in the united states between 1955 and 1961 simian vacuolating virus 40, known as sv40, is a virus found in monkeys and humans, which has the potential. Factors influencing the production of recombinant sv40 vectors. Amazingly, sv40 only needs one protein, the tantigen, to control both of these processes. Transient and stable gene expression in mammalian cells. Sv40 viral particles were detected in both pancreatic islets and acini. In pancreatic islets, all islet cell types were infected by sv40, albeit the infection rate of glucagonproducing.
Sv40 was present in cancers of people who either had or had not received the polio vaccines that were contaminated with sv40. The university of iowa viral vector core facility is supported in part by the university of iowa holden comprehensive cancer center. Viral vector core facility 500 newton road 221 eckstein medical research building iowa city, ia 52242 phone. Simian virus 40 sv40, a possible human polyomavirus. Here, qualification of the relevant manufacturers, good supply chain control, and additional testing contribute to another level of complexity. In the past, simian virus 40 sv40 has been used as a cloning vehicle to clone foreign genes by substituting portions of the viral genome vital for viral replication. Wilson1 abstract breast cancer brain metastases are a deadly sequela of. Lentiviral vector that expresses nuclear localization signal nlscontaining cre recombinase from internal cmv promoter. For example the replication of sv40 dna requires a single viral protein, the large tantigen, whereas the other factors are supplied by the host cell.
These findings were supported by the isolation of infectious sv40 from a choroid plexus tumor. Sv40, the prototype of polyomavirus, has been extensively studied as a model of dna tumor virus, since sv40 induces tumor in rodents and it can be readily propagated in cultured cells. Vector manufacturing and testing for gene and cell therapy. Consequently, we devised a gene transfer system based on simian virus 40 sv40 1 as a vector.
Although viral vectors are occasionally created from pathogenic viruses. To detect vector dna peak fractions for sv40, jcv, bkv, and lpv capsid vector particles, 1. Dk173592b1 recombinant viral expression vector capable of. Infection with sv40 or transfection with vectors carrying the gene coding for the sv40 large tantigen can lead to immortalization of primary cells and can induce tumor formation in animals. However, the sponsors should take the following steps to resolve questions raised by human exposure to sv40 in viral vaccines prepared in rhesus monkey cells in the 1950s and 1960s. Yet the viral vector manufacturer does not directly control production of these materials. However, transiently transfected vectors as well as the presence of sv40 virus large tantigen tag cause serious technical and safety considerations. Sv40 vectors carrying minimal sequence of viral origin with. People with cancers who were born after sv40 was no longer a contaminant of the polio vaccine were found to have evidence for sv40 in their cancerous cells. Viral vectors are tailored to their specific applications but generally share a few key properties. Efficient expression of small rna polymerase iii genes from a. Simian virus 40 sv40 is a small dna virus that was the first animal virus for which the complete genome was sequenced.
Although viral vectors are occasionally created from pathogenic viruses, they are modified in such a way as to minimize the risk of handling them. This is a free resource for the scientific community that is compiled. The present invention provides sv40based viral vectors for use in the delivery of genes to target cells. Sv40 is an abbreviation for simian vacuolating virus 40 or simian virus 40, a polyomavirus that is found in both monkeys and humans. Use of sv40 recombinant viruses as vectors for gene. Pancreatic acinar and islet cell infection by lowdose sv40. A novel sv40based vector successfully transduces and expresses an alpha 1antitrypsin ribozyme in a human hepatomaderived cell line gene therapy 1999 6.
Use of sv40 recombinant viruses as vectors for gene therapy. Pdf most gene therapy approaches employ viral vectors for gene delivery. The rationale for this approach was based on several important aspects of sv40 virus and its. Sv40 has not been present in any vaccine since 1963. Thomas robertss lab contains the insert sv40 lt and is published in cancer cell. The rationale for this approach was based on several important aspects of sv40 virus and its activity. Simian virus 40 vectors for pulmonary gene therapy. Sv40derived vectors provide effective transgene expression. We report here successful immunization using such an sv40derived viral vector. The cterminal region of lt is required for efficient viral gene expression and replication in the african green monkey kidney agmk cv1p cell line. Also, evidence indicates that zoonotic transmission of sv40 should be a. Ideally, these vectors should be produced at high titer and purity with. Adenoassociated virus shuttle plasmids viral vector core.
Pdf the simian virus 40 polyadenylation signal sv40 polya has been routinely inserted downstream of the polyhedrin promoter in many baculovirus. The unique combination of multiple viral systems provides for vectors with increased carrying capacity. This system combines efficient gene delivery of the sv40 viral vector with the presumed safety of nonviral vectors. Testing of retroviral vectorbased human gene therapy products for replication competent retrovirus during product manufacture and patient followup draft guidance for industry july 2018. The virusinfected cells formed colonies 34 weeks after infection in medium containing fetal calf. Sv40 lt cterminal region increases viral gene expression in monkey and human cells. Thus, the mechanism of viral entry to deliver foreign genes is exploited, whereas the. Most gene therapy approaches employ viral vectors for gene delivery. Here, qualification of the relevant manufacturers, good supply chain control, and additional testing.
Pdf the influence of sv40 polya on gene expression of. Use of sv40based vectors to transduce foreign genes to. Sv40 as an effective gene transfer vector in vivo journal of. Selection of the bestsuited vector is pivotal and requires focused indepth knowledge of the delivery systems and their performances. Pdf factors influencing the production of recombinant sv40.
The vector of any one of claims, wherein the viral vector is the sv40 vector. Sv40, we recently conducted a large, longterm retrospective followup study of birth cohorts exposed or unexposed to sv40 contaminated poliovirus vaccine strickler et al. Schematic diagram of viral constructs with and without sv40 polya signals for egfp gene expression under the acmnpv polh promoter control at the polh, egt and gp37 loci of acmnpv. Sv40 promoter for highlevel constitutive expression of your gene of interest cmv promoter for highlevel constitutive expression of the cycle 3 gfpzeocin fusion. Vaccine contamination with sv40 occurred in the united states between 1955 and 1961 simian vacuolating virus 40, known as sv40, is a virus found in monkeys and humans, which has the potential to cause cancer, mainly in animals, although research indicates that it doesnt cause cancer in humans. In this study, to determine whether rsv40 follows the same route as the wildtype sv40, we generated an rsv40 that is deleted of all viral coding sequences and expresses e.
Also, the sv40 intron induced higher level of epo expression than ivs intron in transfected cho cell. A comparative analysis of constitutive promoters located in. High cloning capacity of in vitro packaged sv40 vectors with. The simian virus 40 polyadenylation signal sv40 polya has been routinely inserted downstream of the polyhedrin promoter in many baculovirus expression vector systems bevs. The present invention provides sv40 based viral vectors for use in the delivery of genes to target cells. Pbmc using two different sv40derived viral vectors. Recombinant viral vector biosafety levels stanford. The unique combination of multiple viral systems provides for vectors with increased carrying capacity and extended host range when compared to normal sv40 vectors. Hundreds of millions of people were immunized with sv40 contaminated vaccine, making this by far the largest singlesource. Hepatitis b surface antigen hbsag cdna was cloned downstream of two tandem sv40 early promoters to yield a t. In this study, aav vectors, in which enhanced gfp expression was directed by one of the six constitutive promoters human. Sv40 infects a wide range of cell types from humans and other mammals, and expresses its genes in them.
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