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Article

Design, Characterization, and Evaluation of scFvCD133/rGelonin: A CD133-Targeting Recombinant Immunotoxin for Use in Combination with Photochemical Internalization

1
Department of Radiation Biology, Institute for Cancer Research, Oslo University Hospital, The Norwegian Radium Hospital, N-0310 Oslo, Norway
2
Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
3
Department of Therapeutic Radiology-Radiation Oncology, University of Minnesota, Masonic Cancer Center, Minneapolis, MN 55455, USA
*
Author to whom correspondence should be addressed.
J. Clin. Med. 2020, 9(1), 68; https://doi.org/10.3390/jcm9010068
Received: 15 November 2019 / Revised: 17 December 2019 / Accepted: 22 December 2019 / Published: 26 December 2019
(This article belongs to the Special Issue The Past, Present and Future of Photodynamic Therapy for Cancers)
The objective of this study was to develop and explore a novel CD133-targeting immunotoxin (IT) for use in combination with the endosomal escape method photochemical internalization (PCI). scFvCD133/rGelonin was recombinantly constructed by fusing a gene (scFvCD133) encoding the scFv that targets both non-glycosylated and glycosylated forms of both human and murine CD133/prominin-1 to a gene encoding the ribosome-inactivating protein (RIP) gelonin (rGelonin). RIP-activity was assessed in a cell-free translation assay. Selective binding and intracellular accumulation of scFvCD133/rGelonin was evaluated by flow cytometry and fluorescence microscopy. PCI of scFvCD133/rGelonin was explored in CD133high and CD133low cell lines and a CD133neg cell line, where cytotoxicity was evaluated by the MTT assay. scFvCD133/rGelonin exhibited superior binding to and a higher accumulation in CD133high cells compared to CD133low cells. No cytotoxic responses were detected in either CD133high or CD133low cells after 72 h incubation with <100 nM scFvCD133/rGelonin. Despite a severe loss in RIP-activity of scFvCD133/rGelonin compared to free rGelonin, PCI of scFvCD133/rGelonin induced log-fold reduction of viability compared to PCI of rGelonin. Strikingly, PCI of scFvCD133/rGelonin exceeded the cytotoxicity of PCI of rGelonin also in CD133low cells. In conclusion, PCI promotes strong cytotoxic activity of the per se non-toxic scFvCD133/rGelonin in both CD133high and CD133low cancer cells. View Full-Text
Keywords: cancer stem cells; CD133; prominin-1; photochemical internalization; photodynamic therapy; immunotoxins; ribosome-inactivating proteins; drug delivery cancer stem cells; CD133; prominin-1; photochemical internalization; photodynamic therapy; immunotoxins; ribosome-inactivating proteins; drug delivery
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MDPI and ACS Style

Olsen, C.E.; Cheung, L.H.; Weyergang, A.; Berg, K.; Vallera, D.A.; Rosenblum, M.G.; Selbo, P.K. Design, Characterization, and Evaluation of scFvCD133/rGelonin: A CD133-Targeting Recombinant Immunotoxin for Use in Combination with Photochemical Internalization. J. Clin. Med. 2020, 9, 68. https://doi.org/10.3390/jcm9010068

AMA Style

Olsen CE, Cheung LH, Weyergang A, Berg K, Vallera DA, Rosenblum MG, Selbo PK. Design, Characterization, and Evaluation of scFvCD133/rGelonin: A CD133-Targeting Recombinant Immunotoxin for Use in Combination with Photochemical Internalization. Journal of Clinical Medicine. 2020; 9(1):68. https://doi.org/10.3390/jcm9010068

Chicago/Turabian Style

Olsen, Cathrine E., Lawrence H. Cheung, Anette Weyergang, Kristian Berg, Daniel A. Vallera, Michael G. Rosenblum, and Pål K. Selbo 2020. "Design, Characterization, and Evaluation of scFvCD133/rGelonin: A CD133-Targeting Recombinant Immunotoxin for Use in Combination with Photochemical Internalization" Journal of Clinical Medicine 9, no. 1: 68. https://doi.org/10.3390/jcm9010068

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