Efficient nonviral transfection of dendritic cells and their use for in vivo immunization

Irvine, Alistair S.; Trinder, P. K. E.; Laughton, David L.; Ketteringham, Helen; McDermott, R. H.; Reid, Sophie C.H.; Haines, A. M. R.; Amir, Abdu; Husain, Rhonda; Doshi, Rajeev; Young, Lawrence S.; Mountain, Andrew

doi:10.1038/82383

Cited by 90 publications

(61 citation statements)

References 28 publications

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“…This approach does not appear to afford advantages over the one that we describe and is feasible only if purified Ags are available. Positively charged peptides have also been used to dramatically increase DC transfection efficiency (33). DC transfected with peptide-coated plasmids encoding Ags of interest induced Ab and CTL responses and vaccinated against tumors.…”

Section: Discussionmentioning

confidence: 99%

Dendritic Cells Transduced with Protein Antigens Induce Cytotoxic Lymphocytes and Elicit Antitumor Immunity

Shibagaki

Udey

2002

The Journal of Immunology

124

106

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Dendritic cell (DC)-based vaccines are being developed for treatment of patients with cancer, in part because DC are potent inducers of CD8+ CTL. DC MHC class I:antigenic peptide complexes that are required for CTL elicitation are most often generated by incubating DC with peptides or by transfecting (or transducing) DC with cDNAs (or viral vectors) that encode protein Ags. The former approach is feasible when MHC class I Ags and relevant peptides are known. The latter approach may be hampered by inefficient DC transfection (transduction) and/or difficulties associated with preparation and use of viral vectors. Herein we demonstrate that a bacterial recombinant model tumor-associated Ag (OVA) that contains the HIV TAT protein transduction domain (PTD) was readily engineered and purified, efficiently transduced murine lymphocytes and DC, and was processed by proteasomes for MHC class I-restricted presentation to CTL. In addition, PTD-containing rOVA was processed and presented by DC to CD4 T cells as efficiently as native OVA or rOVA lacking the PTD. PTD-OVA-transduced DC induced CTL in vivo in a Th cell-independent fashion and vaccinated against OVA-expressing tumors. In contrast, rOVA lacking the PTD did not enter the DC MHC class I presentation pathway and DC treated with this protein did not prime OVA-specific CTL in vivo. Treatment of mice harboring clinically apparent OVA-expressing tumors with PTD-OVA-transduced DC resulted in tumor regression in some animals. This straightforward vaccination strategy may translate into DC-based treatments for patients with cancer and other serious diseases.

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Section: Discussionmentioning

confidence: 99%

Dendritic Cells Transduced with Protein Antigens Induce Cytotoxic Lymphocytes and Elicit Antitumor Immunity

Shibagaki

Udey

2002

The Journal of Immunology

124

106

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“…Our recent work shows that the average transfection efficiency of human DC with [CL22] 2 -DNA complexes (17%) is greatly superior to alternative nonviral transfection agents. 30 Such primary DC are generally refractory to transfection. 31,32 Our results with [CL22] 2 -DNA complexes are inconsistent with this hypothesis.…”

Section: Figure 9 Comparison Of Transfection Activity Of [Cl22] 2 -Dnmentioning

confidence: 99%

CL22 – a novel cationic peptide for efficient transfection of mammalian cells

Haines¹,

Irvine²,

Mountain³

et al. 2001

Gene Ther

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Condensing peptide-DNA complexes have great potential as nonviral agents for gene delivery. To date, however, such complexes have given transfection activities greatly inferior to adenovirus and somewhat inferior to cationic lipid-DNA complexes, even for cell lines and primary cells in vitro. We report here the identification of a novel condensing peptide, CL22, which forms DNA complexes that efficiently transfect many cell lines, as well as primary dendritic and endothelial cells. We report studies with sequence and structure vari-

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“…[24][25][26][27] Among the numerous nonviral methods to genetically modify DC, we mention the pulsing of DC with naked RNA or plasmid DNA and lipofection of RNA or DNA. [28][29][30][31][32][33][34][35][36] The different methods can either use the full antigenic spectrum of the tumor or can focus on only one TAA. The former strategy is believed to be more effective in eradicating the tumor, because of a reduced risk that antigen-loss variants will arise and escape the immune response.…”

mentioning

confidence: 99%

Induction of Influenza Matrix Protein 1 and MelanA-specific T lymphocytes in vitro using mRNA-electroporated dendritic cells

et al. 2003

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Genetically modified dendritic cells (DC) constitute a promising approach in cancer immunotherapy. Viral gene delivery systems have been shown to be very efficient strategies, but safety concerns for their clinical use in immunotherapy remain an important issue. Recently, the technique of mRNA electroporation was described as a very efficient tool for the genetic modification of human monocyte-derived DC. Here, we show that transgene expression can be modulated by varying the amount of mRNA used for electroporation. We document that CD40 ligation leads to a significant production of IL-12 by the electroporated DC, although the level of IL-12 production is somewhat lower than for non-or mock-electroporated DC. Furthermore, we show that the electroporated DC can be frozen and thawed without loss of viability or function and that Influenza virus Matrix Protein 1 mRNA electroporated DC are capable of inducing a memory cytotoxic T lymphocyte response and are more potent in doing so than mRNA-pulsed DC. Similar results were obtained with MelanA/MART-1 mRNA electroporated DC. These results clearly indicate that mRNA-electroporated DC represent powerful candidates for use as tumor vaccines and could constitute an improvement compared with vaccines using peptide-pulsed DC.

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Efficient nonviral transfection of dendritic cells and their use for in vivo immunization

Cited by 90 publications

References 28 publications

Dendritic Cells Transduced with Protein Antigens Induce Cytotoxic Lymphocytes and Elicit Antitumor Immunity

Dendritic Cells Transduced with Protein Antigens Induce Cytotoxic Lymphocytes and Elicit Antitumor Immunity

CL22 – a novel cationic peptide for efficient transfection of mammalian cells

Induction of Influenza Matrix Protein 1 and MelanA-specific T lymphocytes in vitro using mRNA-electroporated dendritic cells

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