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Elimination of murine and human T-cell epitopes in recombinant immunotoxin eliminates neutralizing and anti-drug antibodies in vivo

Cellular & Molecular Immunology volume 14, pages 432–442 (2017)Cite this article

Abstract

Antibodies against the toxin portion of recombinant immunotoxins (RIT) reduce their efficacy and pose a potential safety risk. To overcome this problem we mutated the very immunogenic immunotoxin SS1P to produce LMB-T20, a de-immunized RIT that has the eight human T-cell epitopes in SS1P modified or removed. To determine the effect of T-cell epitope removal in vivo we mapped the T-cell epitopes in immune-competent BALB/c mice and found that these mice recognize two epitopes. One corresponds to the human immunodominant T-cell epitope and the other to a human subdominant epitope; both were eliminated in LMB-T20. We found that mice immunized with LMB-T20 did not have T-cell activation and did not develop anti-drug antibodies (ADA), whereas mice immunized with SS1P, showed T-cell activation, and developed ADA detected by both ELISA and drug neutralizing assays. The ability of the mice treated with LMB-T20 to respond to other antigens was not compromised. We conclude that elimination of T-cell epitopes is sufficient to prevent formation of antibodies to an immunogenic foreign protein.

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Acknowledgements

The authors thank Dr. Chin-Hsien Tai for creating the structural model of RIT for figure 1. Supplementary information of this article can be found on the Cellular & Molecular Immunology’s website (http://www.nature.com/cmi).

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Authors and Affiliations

  1. Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Ronit Mazor, Selamawit Addissie, Youjin Jang, Gilad Kaplan & Ira Pastan

  2. Leidos Biomedical Research, Inc., National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Devorah Crown

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  1. Ronit Mazor
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  2. Devorah Crown
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Correspondence to Ira Pastan.

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Supplementary information of this article can be found on the Cellular & Molecular Immunology’s website (http://www.nature.com/cmi).

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Mazor, R., Crown, D., Addissie, S. et al. Elimination of murine and human T-cell epitopes in recombinant immunotoxin eliminates neutralizing and anti-drug antibodies in vivo. Cell Mol Immunol 14, 432–442 (2017). https://doi.org/10.1038/cmi.2015.91

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