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

Tuyaerts, Sandra; Michiels, Annelies; Corthals, Jurgen; Bonehill, Aude; Heirman, Carlo; Greef, Catherine De; Noppe, Sofie; Thielemans, Kris

doi:10.1038/sj.cgt.7700622

Cited by 43 publications

(31 citation statements)

References 43 publications

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“…Therefore, DC were electroporated with mRNA either before or after maturation by a previously optimized protocol yielding a transfection efficiency of up to 80%. 40 We here show that the transgene expression by DC electroporated with eGFP mRNA either before or after maturation is high, even at 120 h after electroporation. At 4 h after transfection, we observed a significantly higher transfection efficiency of DC electroporated in a mature state when compared to DC electroporated in an immature state.…”

Section: Discussionmentioning

confidence: 60%

“…40 As described in Materials and methods, DC were electroporated on day 6 of DC culture with eGFP or tNGFR encoding mRNA and matured with the inflammatory cytokine cocktail either 24 h before or 4 h after electroporation ( Figure 1 for the timing of the different manipulations of the cells). The percentage of transgene-expressing cells was equal when the cells were transfected with either eGFP or tNGFR encoding mRNA (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…Several recent reports have indicated that loading of immature DC with whole tumor-derived mRNA or defined mRNA represents an effective nonviral strategy to stimulate T-cell responses both in in vitro and in vivo models. [32][33][34][35][36][38][39][40]42 In this study, we have used the recently described electroporation method to introduce defined mRNA into DC. Many factors may play an important role in the final outcome of this method.…”

Section: Discussionmentioning

confidence: 99%

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Electroporation of immature and mature dendritic cells: implications for dendritic cell-based vaccines

et al. 2005

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Until now, studies utilizing mRNA electroporation as a tool for the delivery of tumor antigens to human monocyte-derived dendritic cells (DC) have focused on DC electroporated in an immature state. Immature DC are considered to be specialized in antigen capture and processing, whereas mature DC present antigen and have an increased T-cell stimulatory capacity. Therefore, the consensus has been to electroporate DC before maturation. We show that the transfection efficiency of DC electroporated either before or after maturation was similarly high. Both immature and mature electroporated DC, matured in the presence of an inflammatory cytokine cocktail, expressed mature DC surface markers and preserved their capacity to secrete cytokines and chemokines upon CD40 ligation. In addition, both immature and mature DC can be efficiently cryopreserved before or after electroporation without deleterious effects on viability, phenotype or T-cell stimulatory capacity including in vitro antigen-specific T-cell activation. However, DC electroporated after maturation are more efficient in in vitro migration assays and at least as effective in antigen presentation as DC electroporated before maturation. These results are important for vaccination strategies where an optimal antigen presentation by DC after migration to the lymphoid organs is crucial. Gene Therapy (2005) 12, 772-782.

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Section: Discussionmentioning

confidence: 60%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Electroporation of immature and mature dendritic cells: implications for dendritic cell-based vaccines

et al. 2005

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“…23 pGEM-4Z/sig-MART1-LAMP1/A64 encodes the chimeric MART1 protein carrying the peptide signal and lysosomal sorting signal of human LAMP1 protein, which have high homology with those of mice. 24,25 pGEM-4Z/LUC/A64 encoding the firefly Luciferase gene (LUC) under the T7 RNA polymerase promoter of l-bacteriophage was constructed as described previously.…”

Section: Animals and Cell Linesmentioning

confidence: 99%

mRNA-based cancer vaccine: prevention of B16 melanoma progression and metastasis by systemic injection of MART1 mRNA histidylated lipopolyplexes

et al. 2007

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Immunization with mRNA encoding tumor antigen is an emerging vaccine strategy for cancer. In this paper, we demonstrate that mice receiving systemic injections of MART1 mRNA histidylated lipopolyplexes were specifically and significantly protected against B16F10 melanoma tumor progression. The originality of this work concerns the use of a new tumor antigen mRNA formulation as vaccine, which allows an efficient protection against the growth of a highly aggressive tumor model after its delivery by intravenous route. Synthetic melanoma-associated antigen MART1 mRNA was formulated with a polyethylene glycol (PEG)ylated derivative of histidylated polylysine and L-histidine-(N,N-di-n-hexadecylamine)ethylamide liposomes (termed histidylated lipopolyplexes). Lipopolyplexes comprised mRNA/polymer complexes encapsulated by liposomes. The tumor protective effect was induced with MART1 mRNA carrying a poly(A) tail length of 100 adenosines at an optimal dose of 12.5 mg per mouse. MART1 mRNA lipopolyplexes elicited a cellular immune response characterized by the production of interferon-g and the induction of cytotoxic T lymphocytes. Finally, the anti-B16 response was enhanced using a formulation containing both MART1 mRNA and MART1-LAMP1 mRNA encoding the antigen targeted to the major histocompatibility complex class II compartments by the lysosomal sorting signal of LAMP1 protein. Our results provide a basis for the development of mRNA histidylated lipopolyplexes for cancer vaccine.

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“…10,[18][19][20][21][22] By our strategy, we intend to make the DCs present a wide spectrum of antigens that are relevant to each patient's tumor. The tumor-RNA approach overcomes the requirement for defined human lymphocyte antigen (HLA) alleles and for expression of identified antigens by the tumors.…”

Section: Introductionmentioning

confidence: 99%

Phase I/II trial of melanoma therapy with dendritic cells transfected with autologous tumor-mRNA

et al. 2006

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We have developed an individualized melanoma vaccine based on transfection of autologous dendritic cells (DCs) with autologous tumor-mRNA. Dendritic cells loaded with complete tumor-mRNA may generate an immune response against a broad repertoire of antigens, including unique patient-specific antigens. The purpose of the present phase I/II trial was to evaluate the feasibility and safety of the vaccine, and the ability of the DCs to elicit T-cell responses in melanoma patients. Further, we compared intradermal (i.d.) and intranodal (i.n.) vaccine administration. Twenty-two patients with advanced malignant melanoma were included, each receiving four weekly vaccines. Monocyte-derived DCs were transfected with tumor-mRNA by electroporation, matured and cryopreserved. We obtained successful vaccine production for all patients elected. No serious adverse effects were observed. A vaccine-specific immune response was demonstrated in 9/19 patients evaluable by T-cell assays (T-cell proliferation/interferon-g ELISPOT) and in 8/18 patients evaluable by delayed-type hypersensitivity (DTH) reaction. The response was demonstrated in 7/10 patients vaccinated intradermally and in 3/12 patients vaccinated intranodally. We conclude that immuno-gene-therapy with the described DC-vaccine is feasible and safe, and that the vaccine can elicit in vivo T-cell responses against antigens encoded by the transfected tumor-mRNA. The response rates do not suggest an advantage in applying i.n. vaccination. Cancer Gene Therapy (2006) 13, 905-918.

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

Cited by 43 publications

References 43 publications

Electroporation of immature and mature dendritic cells: implications for dendritic cell-based vaccines

Electroporation of immature and mature dendritic cells: implications for dendritic cell-based vaccines

mRNA-based cancer vaccine: prevention of B16 melanoma progression and metastasis by systemic injection of MART1 mRNA histidylated lipopolyplexes

Phase I/II trial of melanoma therapy with dendritic cells transfected with autologous tumor-mRNA

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