Abstract
CD19 is a B-lineage-specific transmembrane signaling protein participating in the control of proliferation and differentiation. It is present at high surface density on chronic B-lymphocytic leukemia (B-CLL) cells and cells of other B-cell malignancies, and is a prime target for therapy with antibody-derived agents. Many attempts have been made to target malignant cells via CD19, but to date none of these agents have received drug approval. Here we report the design of a monovalent immunotoxin consisting of a CD19-specific single-chain Fv antibody fragment fused to a derivative of Pseudomonas Exotoxin A. This fusion protein induced efficient antigen-restricted apoptosis of several human leukemia- and lymphoma-derived cell lines including Nalm-6, which it eliminated at an effective concentration (EC50) of 2.5 nM. The agent displayed synergistic toxicity when used in combination with valproic acid and cyclosporin A in cell-culture assays. It induced apoptosis of primary malignant cells in 12/12 samples from B-CLL patients, including patients responding poorly to fludarabine, and of cells from one pediatric acute lymphoblastic leukemia patient. In NOD/SCID mice transplanted with Nalm-6 cells, the toxin prevented engraftment and significantly prolonged survival of treated mice. Owing to its efficient antigen-restricted antileukemic activity, the agent deserves further development towards clinical testing.
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Acknowledgements
We thank Dr W Wels for the Pseudomonas ETA cDNA, and Dr Torsten Haferlach for determining the p53 status of the B-CLL patients. We also thank Th Lange for administrative assistance. This study was supported by research grant 2003.015.1 from the Wilhelm Sander Stiftung, Neustadt, Germany; Association of Supporters of the University Children's Hospital, Erlangen, Germany; Association ‘Kaminkehrer helfen krebskranken Kindern’, Untersiemau, Germany (to GHF); Beitlich Foundation, Tuebingen, Germany (to PL and GHF); Research Training Grant GK592 from the Deutsche Forschungsgemeinschaft; intramural funding from the University of Erlangen's ELAN program; stipends from the Sofie Wallner-Stiftung, Erlangen, Germany (to MS); Schickedanz KinderKrebs Stiftung, Fuerth, Germany (to JS).
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Schwemmlein, M., Stieglmaier, J., Kellner, C. et al. A CD19-specific single-chain immunotoxin mediates potent apoptosis of B-lineage leukemic cells. Leukemia 21, 1405–1412 (2007). https://doi.org/10.1038/sj.leu.2404687
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DOI: https://doi.org/10.1038/sj.leu.2404687
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