A new way to generate cytolytic tumor-specific T cells: electroporation of RNA coding for a T cell receptor into T lymphocytes

Schaft, Niels; Dörrie, Jan; Müller, Ina; Beck, Verena; Baumann, Stefanie; Schunder, Tanja; Kämpgen, Eckhart; Schuler, Gerold

doi:10.1007/s00262-005-0098-2

Cited by 93 publications

(128 citation statements)

References 30 publications

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“…32,33 Subsequently, RNA electroporation was used to transfer full-length TCR to T cells to redirect them with a new specificity. 9,11,25,26 Owing to the fact that RNA does not integrate into the genome and is quickly degraded, this technique has certain advantages over retroviral transduction.…”

Section: Discussionmentioning

confidence: 99%

“…25 Briefly, T cells were washed once with pure RPMI 1640 and once with OptiMEM without phenol red (Lonza, Vervier, Belgium; all at room temperature), and resuspended in OptiMEM at a concentration of up to 14 Â 10 7 cells per ml. RNA was transferred to a 4-mm cuvette (Peqlab, Erlangen, Germany; 150 mg ml À1 final concentration), 100-600 ml of cell suspension was added and pulsed in a Genepulser Xcell (Bio-Rad, Munich, Germany).…”

Section: Rna Electroporation Of Cd4mentioning

confidence: 99%

“…9,11,25,26 Owing to the limited half-life and the lack of stable genomic integration, RNA-based immunoreceptor expression is transient. If an autoreactive T cell is unintendedly generated, the autoaggressive pathology will be limited due to the short half-life of the immunoreceptor expression, without the need of therapeutic T-cell elimination.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+ and CD8+ T cells for use in the adoptive immunotherapy of cancer

et al. 2009

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Section: Discussionmentioning

confidence: 99%

Section: Rna Electroporation Of Cd4mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+ and CD8+ T cells for use in the adoptive immunotherapy of cancer

et al. 2009

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“…The CD4 + cells were then cultivated overnight in MLPC medium with 20 IU IL-7/ml (PeproTech), as noted in Schaft et al (2006), and used for the antigen-specific CD4 + T-cell-stimulation assays. CD4 + T-cells to be used for mixed-lymphocyte reactions (MLR) were isolated with a MACS isolation kit (CD4 + T-Cell Isolation Kit II, human; Miltenyi) and cultivated in MLPC medium without IL-7 to avoid T-cell-stimulation signals.…”

Section: Generation Of Human Cd4mentioning

confidence: 99%

Norm- and hypo-fractionated radiotherapy is capable of activating human dendritic cells

Kulzer¹,

Rubner²,

Deloch³

et al. 2014

Journal of Immunotoxicology

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Despite the transient immunosuppressive properties of local radiotherapy (RT), this classical treatment modality of solid tumors is capable of inducing immunostimulatory forms of tumor-cell death. The resulting 'immunotoxicity' in the tumor, but not in healthy tissues, may finally lead to immune-mediated destruction of the tumor. However, little is known about the best irradiation scheme in this setting. This study examines the immunological effects of differently irradiated human colorectal tumor cells on human monocyte-derived dendritic cells (DC). Human SW480 tumor cells were irradiated with a norm-fractionation scheme (5 Â 2 Gy), a hypo-fractionated protocol (3 Â 5 Gy), and with a high single irradiation dose (radiosurgery; 1 Â 15 Gy). Subsequently, human immature DC (iDC) were co-incubated with supernatants (SN) of these differently treated tumor cells. Afterwards, DC were analyzed regarding the expression of maturation markers, the release of cytokines, and the potential to stimulate CD4 + T-cells. The co-incubation of iDC with SN of tumor cells exposed to norm-or hypo-fractionated RT resulted in a significantly increased secretion of the immune activating cytokines IL-12p70, IL-8, IL-6, and TNF, compared to iDC co-incubated with SN of tumor cells that received a high single irradiation dose or were not irradiated. In addition, DC-maturation markers CD80, CD83, and CD25 were also exclusively elevated after co-incubation with the SN of fractionated irradiated tumor cells. Furthermore, the SN of tumor cells that were irradiated with norm-or hypo-fractionated RT triggered iDC to stimulate CD4 + T-cells not only in an allogenic, but also in an antigen-specific manner like mature DC. Collectively, these results demonstrate that norm-and hypo-fractionated RT induces a fast human colorectal tumor-cell death with immunogenic potential that can trigger DC maturation and activation in vitro. Such findings may contribute to the improvement of irradiation protocols for the most beneficial induction of anti-tumor immunity.

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“…40,41 Subsequently, RNA electroporation was used to transfer full-length TCR to T cells to confer upon them a new specificity. [42][43][44] Using this method, we previously showed that RNA-mediated gene transfer redirection of T-cell target antigens is an efficient nonviral gene transfer technology for T lymphocytes in terms of transduction efficiency and convenience in preparation. In this study, IL-2-PBL and CIR/IL-2-PBL highly expressed IL-2.…”

Section: Discussionmentioning

confidence: 99%

Direct and indirect antitumor effects by human peripheral blood lymphocytes expressing both chimeric immune receptor and interleukin-2 in ovarian cancer xenograft model

Lee

et al. 2010

Cancer Gene Ther

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Human peripheral blood lymphocytes (PBLs) electroporated with RNA encoding anti-Her-2/neu-specific chimeric immune receptor (CIR) have been reported to elicit potent immune responses against SKOV3 tumors in a nude mouse model. However, CIRelectroporated PBL (CIR-PBL) did not proliferate, and the cell number rapidly decreased in the absence of exogenous interleukin-2 (IL-2). In this study, PBLs electroporated with both CIR and IL-2 RNA (CIR/IL-2-PBL) were studied to determine whether antitumor effects could be improved by adoptive immunotherapy. CIR and IL-2 were expressed in CIR/IL-2-PBL at levels similar to PBLs electroporated, with IL-2 RNA (IL-2-PBL) or CIR-PBL. Transfer of IL-2 RNA induced proliferation and prolonged survival of PBLs in vitro. In a xenograft model, both IL-2-PBL and CIR/IL-2-PBL showed significantly higher antitumor effects than CIR-PBL. The number of tumor-infiltrating natural killer (NK) cells was significantly increased in IL-2-PBL and CIR/IL-2-PBL. After NK cell depletion, IL-2-PBL showed significantly lower antitumor effects than CIR/IL-2-PBL. These results suggest that transfer of IL-2 RNA to CIR-PBL can promote NK cell infiltration of tumors and prolong survival of infused PBLs in vivo. RNA electroporated PBLs may represent efficient tools for delivery of functional molecules to tumors by multiple gene transfer.

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A new way to generate cytolytic tumor-specific T cells: electroporation of RNA coding for a T cell receptor into T lymphocytes

Cited by 93 publications

References 30 publications

Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+ and CD8+ T cells for use in the adoptive immunotherapy of cancer

Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+ and CD8+ T cells for use in the adoptive immunotherapy of cancer

Norm- and hypo-fractionated radiotherapy is capable of activating human dendritic cells

Direct and indirect antitumor effects by human peripheral blood lymphocytes expressing both chimeric immune receptor and interleukin-2 in ovarian cancer xenograft model

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