Transiently redirected T cells for adoptive transfer

Almåsbak, Hilde; Rian, Edith; Hoel, Hanna Julie; Pulé, Martin; Wälchli, Sébastien; Kvalheim, Gunnar; Gaudernack, Gustav; Rasmussen, Anne Marie

doi:10.3109/14653249.2010.542461

Cited by 61 publications

(62 citation statements)

References 38 publications

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“…CAR transfection efficiency was similar in dividing and non-dividing cells (Fig. 1B-C signaling motifs for engineering of T cells (26). To investigate the effects of these structural modifications on CAR expression and transfection efficacy, HA21 was compared to a series of alternative constructs in terms of hinge-spacer length and signaling motifs shown in Fig.…”

Section: Optimizing Electroporation Of Car Mrna Into Primary Human Nkmentioning

confidence: 99%

“…Previously described transfection protocol was adapted for the optimized CAR expression in NK cells (26). Cytokine stimulated primary NK cells or B cell-depleted PBMC were washed and resuspended in serum-free SCGM medium at 10-50 × 10 6 cells/ml.…”

Section: Car Mrna Electroporationmentioning

confidence: 99%

“…For the purpose of this study both constructs were recloned from the retrovirus vector SFG into the mRNA expression vector pCIpA102 as described in (25). In vitro mRNA synthesis was performed as previously described (26). Anti-Reverse Cap Analog (Trilink Biotechnologies Inc.) was used to cap the RNA.…”

Section: Vectors Construction and In Vitro Mrna Synthesismentioning

confidence: 99%

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Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors

Oei

Siernicka

Graczyk-Jarzynka

et al. 2018

Cancer Immunology Research

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Section: Optimizing Electroporation Of Car Mrna Into Primary Human Nkmentioning

confidence: 99%

Section: Car Mrna Electroporationmentioning

confidence: 99%

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Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors

Oei

Siernicka

Graczyk-Jarzynka

et al. 2018

Cancer Immunology Research

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“…This was done either by mRNA electroporation or retroviral transduction as previously described. 36,37 T cells were coincubated with DCs at a ratio of 1:2 for 5 hours in the presence of GolgiPlug and GolgiStop (both BD Biosciences) before intracellular staining was performed as described earlier. All conditions were tested in duplicate, and the experiment was repeated 2-3 times.…”

Section: Proliferation Testsmentioning

confidence: 99%

“…T cells were expanded from peripheral blood mononuclear cells as previously described 36 using Dynabeads ClinExVivo CD3/CD28 (Life Technologies). Expanded T cells (day 10 postactivation) were transfected with the DMF5 TCR (a kind gift of Dr R. Morgan, National Cancer Institute, Bethesda).…”

Section: Proliferation Testsmentioning

confidence: 99%

Enhancing Antitumor Immune Responses by Optimized Combinations of Cell-penetrating Peptide-based Vaccines and Adjuvants

Belnoue¹,

Berardino-Besson

Gaertner

et al. 2016

Molecular Therapy

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Cell penetrating peptides (CPPs) from the protein ZEBRA are promising candidates to exploit in therapeutic cancer vaccines, since they can transport antigenic cargos into dendritic cells and induce tumor-specific T cells. Employing CPPs for a given cancer indication will require engineering to include relevant tumor-associated epitopes, administration with an appropriate adjuvant, and testing for antitumor immunity. We assessed the importance of structural characteristics, efficiency of in vitro transduction of target cells, and choice of adjuvant in inducing the two key elements in antitumor immunity, CD4 and CD8 T cells, as well as control of tumor growth in vivo. Structural characteristics associated with CPP function varied according to CPP truncations and cargo epitope composition, and correlated with in vitro transduction efficiency. However, subsequent in vivo capacity to induce CD4 and CD8 T cells was not always predicted by in vitro results. We determined that the critical parameter for in vivo efficacy using aggressive mouse tumor models was the choice of adjuvant. Optimal pairing of a particular ZEBRA-CPP sequence and antigenic cargo together with adjuvant induced potent antitumor immunity. Our results highlight the irreplaceable role of in vivo testing of novel vaccine constructs together with adjuvants to select combinations for further development.

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Optimization of electroporation and other non‐viral gene delivery strategies for T cells

Harris

Elmer

2020

Biotechnology Progress

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CART therapy is a particularly effective treatment for some types of cancer that uses retroviruses to deliver the gene for a chimeric antigen receptor (CAR) to a patient's T cells ex vivo. The CAR enables the T cells to bind and eradicate cells with a specific surface marker (e.g., CD19 + B cells) after they are transfused back into the patient. This treatment was proven to be particularly effective in treating non-Hodgkin's lymphoma (NHL) and acute lymphoblastic leukemia (ALL), but the current CART cell manufacturing process has a few significant drawbacks. For example, while lentiviral and gammaretroviral transduction are both relatively effective, the process of producing viral vectors is time-consuming and costly. Additionally, patients must undergo follow up appointments for several years to monitor them for any unanticipated side effects associated with the virus. Therefore, several studies have endeavored to find alternative non-viral gene delivery methods that are less expensive, more precise, simple, and safe. This review focuses on the current state of the most promising non-viral gene delivery techniques, including electroporation and transfection with cationic polymers or lipids.

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Transiently redirected T cells for adoptive transfer

Cited by 61 publications

References 38 publications

Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors

Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors

Enhancing Antitumor Immune Responses by Optimized Combinations of Cell-penetrating Peptide-based Vaccines and Adjuvants

Optimization of electroporation and other non‐viral gene delivery strategies for T cells

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