Polynucleotide viral vaccines: codon optimisation and ubiquitin conjugation enhances prophylactic and therapeutic efficacy

Liu, Wen Jun; Zhao, Kong‐Nan; Gao, Feng; Leggatt, Graham R.; Fernando, Germain J. P.; Frazer, Ian H.

doi:10.1016/s0264-410x(01)00406-6

Cited by 70 publications

(51 citation statements)

References 42 publications

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“…To test the hypothesis that codon usage bias might determine the differentiation-dependent translation of the L1 genes in cultured KCs, we examined the expression of two heavily codon-substituted L1 genes in differentiating KCs, as these genes, encoding unmodified L1 proteins, have been shown to be expressed well in a wide range of cells in vitro (24,41,69). The Mod L1 genes were each transcribed in both undifferentiated and differentiated KCs ( Fig.…”

Section: Methodsmentioning

confidence: 99%

“…In humans, codon-mediated translational controls may play an important role in the differentiation and regulation of tissue-specific gene products (47). Blockage to translation of PV L1 mRNAs has been overcome by codon modification utilizing mammalian preferred codons without changing the protein sequence (36,41,42,69), but it remains unclear whether codon modification assists L1 protein synthesis by removal of sequences inhibitory to mRNA translation or destabilizing of mRNA or by some other mechanism. Mechanisms postulated to determine instability of L1 mRNA in undifferentiated cells (32,54,62) have not been demonstrated to distinguish between undifferentiated and differentiated KCs, the host cells of PV infection.…”

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confidence: 99%

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Gene Codon Composition Determines Differentiation-Dependent Expression of a Viral Capsid Gene in Keratinocytes In Vitro and InVivo

Zhao

Fang

et al. 2005

Molecular and Cellular Biology

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By establishing mouse primary keratinocytes (KCs) in culture, we were able, for the first time, to express papillomavirus major capsid (L1) proteins by transient transfection of authentic or codon-modified L1 gene expression plasmids. We demonstrate in vitro and in vivo that gene codon composition is in part responsible for differentiation-dependent expression of L1 protein in KCs. L1 mRNA was present in similar amounts in differentiated and undifferentiated KCs transfected with authentic or codon-modified L1 genes and had a similar half-life, demonstrating that L1 protein production is posttranscriptionally regulated. We demonstrate further that KCs substantially change their tRNA profiles upon differentiation. Aminoacyl-tRNAs from differentiated KCs but not undifferentiated KCs enhanced the translation of authentic L1 mRNA, suggesting that differentiation-associated change to tRNA profiles enhances L1 expression in differentiated KCs. Thus, our data reveal a novel mechanism for regulation of gene expression utilized by a virus to direct viral capsid protein expression to the site of virion assembly in mature KCs. Analysis of two structural proteins of KCs, involucrin and keratin 14, suggests that translation of their mRNAs is also regulated, in association with KC differentiation in vitro, by a similar mechanism.Direction of gene expression to undifferentiated or differentiated cells is classically determined by altered promoter methylation or by production of specific transcription factors or posttranscriptionally by interaction of regulatory mRNA sequences with translational regulators (38). Papillomaviruses (PVs) are a family of double-stranded DNA viruses which replicate exclusively in epithelium, promote cell growth, and affect cellular differentiation, giving rise to benign tumors with, for some virus types, potential for malignancy. mRNA encoding the PV major capsid L1 protein can be transcribed from L1 gene expression plasmids in many types of mammalian cells (20). However, translation of the transcribed mRNA to L1 protein is limited in vivo to differentiated keratinocytes (KCs) (5, 53) and to yeast cells (50,67). Although inhibitory mechanisms have been proposed to explain the blockage of PV L1 gene translation in undifferentiated cells (10, 11), no inhibitory factors have been identified as specific for epithelial cells in vitro or in vivo. Thus, the mechanism for the tight differentiation-specific translation of the PV L1 gene in KCs remains to be determined.PVs, like many mammalian DNA viruses, use relatively few "mammalian consensus" codons to encode their capsid genes, manifesting a high AϩT genome content due to third-nucleotide bias to AϩT (68). In humans, codon-mediated translational controls may play an important role in the differentiation and regulation of tissue-specific gene products (47). Blockage to translation of PV L1 mRNAs has been overcome by codon modification utilizing mammalian preferred codons without changing the protein sequence (36,41,42,69), but it remains unclear whether co...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Gene Codon Composition Determines Differentiation-Dependent Expression of a Viral Capsid Gene in Keratinocytes In Vitro and InVivo

Zhao

Fang

et al. 2005

Molecular and Cellular Biology

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“…Construction of a mammalian expression vector based on pcDNA3 and encoding a codon-modified 6bL1E7I gene, which comprises codon-modified HPV6bL1 truncated of the C-terminal 33 nt triplets fused to a gene encoding the first 50 aa of HPV6bE7, has been described elsewhere (25).…”

Section: Construction Of Codon-modified Hpv6bl1e7i Plasmid Dnamentioning

confidence: 99%

“…Immunization of mice with HPV6bL1 polynucleotide was undertaken, as described elsewhere (25). Briefly, plasmid was purified using a Qiagen Plasmid Mega Kit (Qiagen, Chatsworth, CA) and dissolved in PBS at a concentration of 1 g/l.…”

Section: Dna Immunizationmentioning

confidence: 99%

IL-10 Mediates Suppression of the CD8 T Cell IFN-γ Response to a Novel Viral Epitope in a Primed Host

Liu

Hardy

et al. 2003

The Journal of Immunology

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Priming to Ag can inhibit subsequent induction of an immune response to a new epitope incorporated into that Ag, a phenomenon referred to as original antigenic sin. In this study, we show that prior immunity to a virus capsid can inhibit subsequent induction of the IFN-γ effector T cell response to a novel CD8-restricted antigenic epitope associated with the virus capsid. Inhibition does not involve Ab to the virus capsid, as it is observed in animals lacking B cells. CD8-restricted virus-specific T cell responses are not required, as priming to virus without CTL induction is associated with inhibition. However, IL-10−/− mice, in contrast to IL-10+/+ mice, generate CD8 T cell and Ab responses to novel epitopes incorporated into a virus capsid, even when priming to the capsid has resulted in high titer Ab to the capsid. Furthermore, capsid-primed mice, unable to mount a response to a novel epitope in the capsid protein, are nevertheless able to respond to the same novel epitope delivered independently of the capsid. Thus, inhibition of responsiveness to a novel epitope in a virus-primed animal is a consequence of secretion of IL-10 in response to presented Ag, which inhibits local generation of new CD8 IFN-γ-secreting effector T cells. Induction of virus- or tumor Ag-specific CD8 effector T cells in the partially Ag-primed host may thus be facilitated by local neutralization of IL-10.

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“…Several different approaches have been examined including the attachment of ubiquitin to DNA constructs [6,7], targeting of tumor antigens to centrosomal compartments to enhance MHC class I presentation [8], the attachment to constructs of calreticulin to target HPV antigens to the MHC I pathway [9], optimization of codon usage for enhanced expression and presentation of HPV oncogenes [6,10,11] , utilization of modified genes to decrease the transformation capacity [12][13][14] or enhancement of intercellular spreading in order to increase the number of cells expressing antigen [15]. All of these approaches have demonstrated increased antigen presentation and superior tumor eradication in mouse models.…”

Section: Introductionmentioning

confidence: 99%

The efficacy of a DNA vaccine containing inserted and replicated regions of the E7 gene for treatment of HPV-16 induced tumors

Brinkman

Kast

2007

Vaccine

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A majority of cervical cancers are associated with HPV-16. A DNA vaccine (E7IR) was designed for prophylactic and therapeutic treatment of HPV-16+ tumors containing two repeats of the E7 gene to inactivate transformation and duplicate available epitopes. Mice were vaccinated then tumor challenged, or challenged and then immunized and monitored for tumor volume and survival. Splenocytes were utilized for in vivo CTL assays. The E7IR vaccine demonstrated decreased tumor volume and enhanced survival in prophylactic and therapeutic experiments and improved CTL mediated lysis. The E7IR vaccine shows promise in prevention of tumor formation and elimination of established tumors. KeywordsHuman Papillomavirus (HPV); DNA Vaccine; Cervical Cancer 1.IntroductionCancer of the uterine cervix is the second most common cause of cancer-related deaths in women worldwide. Approximately 510,000 new cases and 288,000 deaths are reported annually [1]. The development of cervical cancer is associated with infection with high risk types of Human Papillomaviruses (HPV). HPV is detected in 99.7% of cervical carcinomas [2], with high risks types HPV-16 and 18 accounting for nearly 70% of cervical cancer cases [3]. In contrast to many other cancers, the fact that cervical cancer is highly associated with a viral infection creates a unique opportunity for therapeutic vaccination against HPV to either prevent cervical cancer once an individual is infected or after the virus has caused cervical cancer. Integration of the viral genome into the host cell genome, a necessary step in cellular transformation, leads to the loss of some genes necessary for a productive viral life cycle and to deregulated or overexpression of the E6 and E7 oncoproteins. Because E7 has been shown to be consistently expressed in HPV-16 positive tumors, it is a logical target in HPV-16 vaccination strategies. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Preclinical studies using DNA based vaccine approaches tend to focus on enhancing presentation of HPV antigens via the MHC class I or MHC class II pathways [5]. Several different approaches have been examined including the attachment of ubiquitin to DNA constructs [6,7], targeting of tumor antigens to centrosomal compartments to enhance MHC class I presentation [8], the attachment to constructs of calreticulin to target HPV antigens to the MHC I pathway [9], optimization of codon usage for enhanced expression and presentation of HPV oncogenes [6,10,11] , utilization of modified genes to decrease the transformation capacity [12][13][14] or enhancement of intercellular s...

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Polynucleotide viral vaccines: codon optimisation and ubiquitin conjugation enhances prophylactic and therapeutic efficacy

Cited by 70 publications

References 42 publications

Gene Codon Composition Determines Differentiation-Dependent Expression of a Viral Capsid Gene in Keratinocytes In Vitro and InVivo

Gene Codon Composition Determines Differentiation-Dependent Expression of a Viral Capsid Gene in Keratinocytes In Vitro and InVivo

IL-10 Mediates Suppression of the CD8 T Cell IFN-γ Response to a Novel Viral Epitope in a Primed Host

The efficacy of a DNA vaccine containing inserted and replicated regions of the E7 gene for treatment of HPV-16 induced tumors

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