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Primary T cells for mRNA-mediated immunotoxin delivery

Gene Therapy volume 25, pages 47–53 (2018)Cite this article

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Abstract

Immune cells become increasingly attractive as delivery system for immunotoxins in cancer therapy to reduce the intrinsic toxicity and severe side effects of chimeric protein toxins. In this study, we investigated the potential of human primary T cells to deliver a secreted immunotoxin through transient messenger RNA (mRNA) transfection. The chimeric protein toxin was directed toward the neovasculature of cancer cells by fusing a truncated version of Pseudomonas exotoxin A (PE38) to human vascular endothelial growth factor (VEGF) and to the single chain variable fragment (scFv) of anti-Her2/neu. Protocols for the transient transfection of human embryonic kidney cells (HEK293) as well as activated primary human T cells were established. Transient transfection with mRNA coding for the immunotoxins e23-PE38, VEGF-PE38 and its attenuated variant VEGF-PE38D yielded efficient expression and secretion. Mass spectrometry analysis endorsed that a fraction of VEGF-PE38D was properly translocated into the endoplasmic reticulum. Furthermore, cytotoxic activity of immunotoxin secreting T cells toward cancer cells was confirmed in co-culture with ovarian adenocarcinoma cells in the presence of a bispecific antibody (bsAb), highlighting the potential of primary T cells for mRNA-mediated immunotoxin delivery.

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Acknowledgements

We kindly thank Dr Niels Schaft (Uniklinik Erlangen) for providing plasmids pGEM-4Z/EGFP and pGEM-4Z, Dr Gerhard Moldenhauer (German Cancer Research Center) for providing the bsAb HEA125xOKT3, and KIST Europe for basic funding and financial support.

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Author notes

  1. A Philippi

    Present address: 3Current address: BioNTech AG, 55131 Mainz, Germany.,

Authors and Affiliations

  1. Environmental safety group, Korea Institute of Science and Technology (KIST Europe), Saarbrücken, Germany

    R Eggers, A Philippi & M O Altmeyer

  2. Department of Biosciences, Molecular and Cell Biology and Center of Human and Molecular Biology (ZHMB), Saarland University, Saarbrücken, Germany

    F Breinig & M J Schmitt

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  1. R Eggers
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  2. A Philippi
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Correspondence to F Breinig or M J Schmitt.

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Eggers, R., Philippi, A., Altmeyer, M. et al. Primary T cells for mRNA-mediated immunotoxin delivery. Gene Ther 25, 47–53 (2018). https://doi.org/10.1038/gt.2017.87

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