Cloning and expression of foreign genes in vaccinia virus, using a host range selection system

Perkus, Marion E.; Limbach, Keith; Paoletti, Enzo

doi:10.1128/jvi.63.9.3829-3836.1989

Cited by 146 publications

(28 citation statements)

References 41 publications

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“…The vaccinia recombinant expressing mouse p53 cDNA, termed Vp53-wt, was constructed as follows: The DNA fragment encoding mouse full-length wild-type p53 (p53-wt) with a single point mutation in the tetramerization domain was excised from the pGEM-p53 vector (kindly provided by Dr Thomas D. Halazonetis, The Wistar Institute) [36] and cloned into the pSC11 transfer vector for vaccinia virus [37]. A recombinant vaccinia virus containing the p53-wt sequence was generated by homologous recombination with vaccinia virus strain Copenhagen using thymidine kinase-negative cells (TK ± ) [38]. The recombinant viruses were characterized for the presence of the p53 sequence by PCR.…”

Section: Methodsmentioning

confidence: 99%

An Experimental Vaccine Expressing Wild‐Type p53 Induces Protective Immunity against Glioblastoma Cells with High Levels of Endogenous p53

Blaszczyk-Thurin

Ertl

2002

Scand J Immunol

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Inoculation of mice with a recombinant vaccinia virus expressing the full-length mouse wild-type p53 protein (Vp53-wt) was shown to induce partial protection against peripheral challenge with a mouse glioblastoma cell line, termed GL261, expressing high levels of nuclear, endogenous wild-type p53.In vivo experiments with knockout (KO) mice and mice treated with depleting doses of antibodies specific to lymphocyte subsets revealed that vaccine efficacy depended on CD4 and CD8 T cells as well as on natural killer (NK) cells. Vp53-wt virus-vaccinated mice that failed to develop tumours upon challenge with a minimal tumourigenic dose of GL261 cells remained completely resistant to further challenge with increased doses of GL261 cells. The efficacy of the Vp53-wt vaccine was improved by adding recombinant mouse interleukin-12 (rIL-12) as an adjuvant at the time of tumour challenge. The induction of T cells to p53 in Vp53-wt virus-immune mice was also demonstrated at the tumour site by immunochemistry and was further confirmed by a delayed-type hypersensitivity response to the p53 protein, although in vitro experiments using splenocytes from vaccinated mice failed to demonstrate CD4 or CD8 T-cell activity to p53.

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Section: Methodsmentioning

confidence: 99%

An Experimental Vaccine Expressing Wild‐Type p53 Induces Protective Immunity against Glioblastoma Cells with High Levels of Endogenous p53

Blaszczyk-Thurin

Ertl

2002

Scand J Immunol

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“…11 The M form is under the control of the vaccinia H6 promoter. 12 The L form is under the control of an entomopox early promoter.…”

Section: Canarypox Constructmentioning

confidence: 99%

Dna Prime/Canarypox Boost—Based Immunotherapy of Chronic Hepatitis B Virus Infection in A Chimpanzee

et al. 2001

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Duck HBV recombinant vaccinia virus has also been used to attempt the termination of chronic infection with this virus in ducks. 6 A temporary drop in surface antigen titer, but no permanent resolution, was observed. Recently, Rollier et al. 7 reported that immunization of chronically infected ducks with DNA encoding HBsAg and PreS determinants resulted in a lowering of viremia and a decrease in hepatic DNA levels.Based on our previous studies on the protective efficacy of DNA-based immunization against HBV in newborn chimpanzees, 8 in the present study we attempted to terminate the HBV chronic carrier state in a chimpanzee. Chronic HBV carriers are rare, as most chimpanzees infected after birth spontaneously resolve HBV infection. One chronically HBV-infected chimpanzee, which had remained chronic after infection with HBV for 12 years, was available to us for this study. We show that immunization with HBsAg-encoding plasmids, followed by boosting with recombinant canarypox encoding HBsAg, PreS-1, and PreS-2, resulted in disappearance of HBV DNA from blood detectable by a quantitative polymerase chain reaction (PCR) assay beginning 1 week after the canarypox booster and persisting for at least 202 weeks after the onset of immunization. A more sensitive nested PCR assay revealed borderline quantities of HBV DNA during the time when HBV DNA was not detectable by the quantitative PCR. Viral clearance in the liver was also established by the decline of covalently closed circular (ccc) HBV DNA that serves as a template for viral transcription. 9 Down-regulation of HBV replication appeared to be mediated by a noncytolytic mechanism that correlated with interferon gamma (IFN-␥) production. MATERIALS AND METHODS Plasmid ConstructsPlasmid pJW-So, which encodes HBsAg, but not Pre-S1 and Pre-S2, was used for the first DNA-based immunization. The recombinant plasmids used in this study were constructed by standard cloning techniques. The pJW-So clone was constructed by PCR from the HBsAg gene of HBV (ayw subtype) corresponding to 2 to 226 amino acids (aa) of S antigen as a Nhe I/Bgl II fragment. The fragment was

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“…Marker rescue, the principle of the selection system presented here, is well established. In a previous report (Perkus et al, 1989), a host range selection system was described that is based on a parental virus lacking the human host range gene K1L, thus being growth re-FIG. 6.…”

Section: Discussionmentioning

confidence: 99%

“…Restoration of the depleted function by an integration vector allows identification of the recombinant clones. Selection criteria are plaque size (Rodriguez and Esteban, 1989), plaque formation (Blasco and Moss, 1995), and host range (Perkus et al, 1989). In the latter system, the host range gene K1L was deleted from the parental virus, impairing viral growth in certain human cell lines.…”

Section: Introductionmentioning

confidence: 99%

Dominant Host Range Selection of Vaccinia Recombinants by Rescue of an Essential Gene

Holzer¹,

Gritschenberger²,

Mayrhofer³

et al. 1998

Virology

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We report the rescue of a defective vaccinia virus, forming the basis for a stringent selection protocol to generate replicating recombinant virus without the need for marker cassettes and selection agents. Plaques of recombinant virus could be isolated solely by their ability to grow in wild-type cells normally supporting the growth of vaccinia virus. All growth-competent clones analyzed contained the gene of interest in the intended genomic locus and displayed foreign gene expression to the same levels as was seen with classical recombinants obtained by insertion into the vaccinia virus thymidine kinase locus. The system is based on a defective vaccinia virus, expressing exclusively early genes, termed eVAC-1, and an insertion plasmid vector providing the essential function, the uracil DNA glycosylase gene. In addition, the defective virus is free of selection and color marker genes, thus also representing a basic vector for the generation of defective recombinants.

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Cloning and expression of foreign genes in vaccinia virus, using a host range selection system

Cited by 146 publications

References 41 publications

An Experimental Vaccine Expressing Wild‐Type p53 Induces Protective Immunity against Glioblastoma Cells with High Levels of Endogenous p53

An Experimental Vaccine Expressing Wild‐Type p53 Induces Protective Immunity against Glioblastoma Cells with High Levels of Endogenous p53

Dna Prime/Canarypox Boost—Based Immunotherapy of Chronic Hepatitis B Virus Infection in A Chimpanzee

Dominant Host Range Selection of Vaccinia Recombinants by Rescue of an Essential Gene

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