Protein delivery using VP22

Bennett, R.; Dalby, Brian

doi:10.1038/nbt0102-20

Cited by 38 publications

(25 citation statements)

References 3 publications

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“…54 The experimental setting and the transfection methods used to introduce the VP22 expressing plasmids into cells could lead to artifacts. In the Flp study and others, the translocation or the biological effect mediated by VP22 were analyzed after transient transfection which could affect the organization of the cell membrane, leading to its permeabilization.…”

Section: Discussionmentioning

confidence: 99%

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

et al. 2004

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The treatment of solid tumors by retroviral delivery of the herpes simplex virus thymidine kinase (TK) followed by ganciclovir (GCV) treatment has so far shown only limited success in patients. One major drawback in this approach is the lack of efficient in vivo gene delivery to cancer cells. Although, the transduction of every single tumor cell is not a requirement since the bystander effect (BE) mediated by gap junctions allows the diffusion of the toxic GCV metabolites from TK-expressing cells toward untransduced cells. To render the TK/GCV approach more potent, and independent of the level of gap junctions, we have tested the efficiency of a TK mutant (TK30) fused to VP22, a herpes simplex protein that seems to be capable of intercellular trafficking. We failed to detect an increase in the BE with cells expressing VP22 fused to TK30 versus cells containing TK30 alone, and this result forced us to reinvestigate the trafficking properties of VP22. Using very sensitive Western blot and fluorescence assays, we were not able to detect the spread of VP22 fused either to TK30 or GFP. These results indicate that VP22 cannot be used as a cargo to translocate TK30 or GFP.

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Section: Discussionmentioning

confidence: 99%

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

et al. 2004

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“…At these low antigen concentrations of 0.1 nM, no enhanced response was observed for the PTD fusion constructs. Given the large amounts of PTD-coupled proteins needed for efficient immune stimulation, [24][25][26][27] this is a coherent result. As pp65 can be used in combination with other viral antigens for the generation of specific cytotoxic T cell against both pp65 and the other antigens, 53 no interference for the induction of cellular immunity by pp65 is expected.…”

Section: Human Cytomegalovirus Protein Pp65 N Scheller Et Almentioning

confidence: 97%

“…[22][23][24][25][26] However, the PTD-mediated uptake, which is initiated by ionic interactions with the cell surface, still requires high protein concentrations. 24,[27][28][29][30][31] As this limits the stimulation of T cells by PTD-coupled proteins in vivo, more efficient delivery systems are desirable.…”

Section: Introductionmentioning

confidence: 99%

Human cytomegalovirus protein pp65: an efficient protein carrier system into human dendritic cells

et al. 2007

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Protein-based immunogens are usually poor inducers of CD8 + T cells. To enhance the induction of CD8 + T cells, one approach is the use of protein immunogens coupled to protein transduction domains (PTDs). These are small cationic peptide sequences that significantly enhance the uptake of fused proteins into dendritic cells (DC) and then mediate their presentation in the context of major histocompatibility complex class I (MHC-I) and MHC-II molecules. One drawback of this system is the high concentrations of PTD-fusion proteins required. Here, we show that proteins fused to the human cytomegalovirus tegument protein pp65were bound with higher efficiency to DCs than those fused to the described PTDs TatPTD and Penetratin. Furthermore, the fusion of pp65 to proteins led to an enhanced uptake of these proteins by DCs. Once taken up, CD4 + and CD8 + memory T cells were strongly stimulated ex vivo demonstrating that pp65 was efficiently processed and presented in the context of both MHC-I and MHC-II. These data make pp65 a promising delivery system to induce cellular immune responses by fused protein vaccines.

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“…[3][4][5][6][7] Thus, linkage to VP22 has permitted to enhance the biological activity of several proteins such as the tumour suppressor p53 8,9 or the products of the suicide genes thymidine kinase 10,11 and cytosine deaminase. 12 It was also used to deliver functional Flp recombinase 13 and to carry SV40 large T antigen to induce cell cycle reentry in terminally differentiated muscle cells. 14 As a result of its spreading property, VP22 has also been used as an adjuvant in DNA vaccination assays.…”

Section: Introductionmentioning

confidence: 99%

Intercellular trafficking and enhanced in vivo antitumour activity of a non-virally delivered P27-VP22 fusion protein

Zavaglia

Eymin

et al. 2003

Gene Ther

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VP22, a structural protein from herpes simplex virus type I, exhibits the unique property of intercellular trafficking. This protein is exported from primary expressing cells and subsequently imported into neighbouring cells. This property is conserved when VP22 is genetically fused to a protein, making it a promising tool to enhance the delivery of a gene product. We chose to study the intercellular transport and biological effect of a fusion protein between the putative tumour suppressor gene p27 Kip1 and VP22. We show that in vitro, P27VP22 is able to spread as efficiently as VP22. Functionality of the P27VP22 protein was demonstrated by its ability to inhibit cyclin/CDK2 complexes activity. In proliferation and clonogenicity assays, transfection with the P27VP22 plasmid resulted in a stronger cell growth inhibition when compared to transfection with the p27 Kip1 vector. In vivo, sub cutaneous tumours established in nude mice were injected with naked DNA encoding P27 or P27VP22. Our results show that P27VP22 can spread in vivo and that injections of the P27VP22 plasmid resulted in a significantly greater antitumour activity than injections of the P27 plasmid. This study confirms the usefulness of VP22-mediated delivery and suggests that P27VP22 may have applications in cancer gene therapy.

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Protein delivery using VP22

Cited by 38 publications

References 3 publications

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

Human cytomegalovirus protein pp65: an efficient protein carrier system into human dendritic cells

Intercellular trafficking and enhanced in vivo antitumour activity of a non-virally delivered P27-VP22 fusion protein

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