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A Cre-inducible diphtheria toxin receptor mediates cell lineage ablation after toxin administration

Nature Methods volume 2, pages 419–426 (2005)Cite this article

Abstract

A new system for lineage ablation is based on transgenic expression of a diphtheria toxin receptor (DTR) in mouse cells and application of diphtheria toxin (DT). To streamline this approach, we generated Cre-inducible DTR transgenic mice (iDTR) in which Cre-mediated excision of a STOP cassette renders cells sensitive to DT. We tested the iDTR strain by crossing to the T cell– and B cell–specific CD4-Cre and CD19-Cre strains, respectively, and observed efficient ablation of T and B cells after exposure to DT. In MOGi-Cre/iDTR double transgenic mice expressing Cre recombinase in oligodendrocytes, we observed myelin loss after intraperitoneal DT injections. Thus, DT crosses the blood-brain barrier and promotes cell ablation in the central nervous system. Notably, we show that the developing DT-specific antibody response is weak and not neutralizing, and thus does not impede the efficacy of DT. Our results validate the use of iDTR mice as a tool for cell ablation in vivo.

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Figure 1: Scheme of the iDTR system.
Figure 2: Generation of ES cells with Cre-inducible DT sensitivity.
Figure 3: DT-mediated ablation of T cells in CD4-Cre/iDTR mice.
Figure 4: Improvement of ablation by longer and more frequent DT application.
Figure 5: DT-mediated ablation of oligodendrocytes in MOGi-Cre/iDTR mice.
Figure 6: DT-specific humoral immunity.

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Acknowledgements

We thank S. Becker and C. Uthoff-Hachenberg for technical assistance. We also thank J. van de Water, I. Parvanova and A. Croxford for critically reading the manuscript. This work was funded by Deutsche Forschungsgemeinschaft grants WA 1601/1-1 to A.W. and BU 1410/1-1 to T.B. T.B. was a fellow of the International Graduate School of Functional Genomics and Genetics at the Institute for Genetics, Cologne. F.L.H. was supported by the Stammbach foundation. S.J. is a Scholar of the Benoziyo Center for Molecular Medicine.

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  1. Ari Waisman

    Present address: I. Medical Department, Johannes Gutenberg-University Mainz, 55131, Mainz, Germany

Authors and Affiliations

  1. Laboratory for Molecular Immunology, Institute for Genetics, University of Cologne, Cologne, D-50931, Germany

    Thorsten Buch, Christine Tertilt, Tobias J A J Heinen, F Thomas Wunderlich & Ari Waisman

  2. Institute of Neuropathology, University Hospital Zurich, Zurich, CH-8091, Switzerland

    Frank L Heppner

  3. Research Department, Kantonal Hospital St. Gallen, St. Gallen, 9007, Switzerland

    Marcel Kremer

  4. Department of Immunology, Weizmann Institute of Science, Rehovot, 76100, Israel

    Steffen Jung

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Supplementary information

Supplementary Fig. 1

Disruption of myelin in the corpus callosum of DT-treated MOGi-cre/iDTR mice. (PDF 2884 kb)

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Buch, T., Heppner, F., Tertilt, C. et al. A Cre-inducible diphtheria toxin receptor mediates cell lineage ablation after toxin administration. Nat Methods 2, 419–426 (2005). https://doi.org/10.1038/nmeth762

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