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Antigen-specific downregulation of T cells by doxorubicin delivered through a recombinant MHC II–peptide chimera

Nature Biotechnology volume 19, pages 142–147 (2001)Cite this article

Abstract

As the number of drugs with potential therapeutic use for T-cell-mediated diseases increases, there is a need to find methods of delivering such drugs to T cells. The major histocompatibility complex (MHC)–peptide complexes are the only antigen-specific ligands for the T-cell receptor (TCR) expressed on T cells, and they may be an appropriate drug delivery system. We engineered a soluble bivalent MHC class II–peptide chimera on the immunoglobulin scaffold (I-Edαβ/Fcγ2a/HA110-120, DEF) that binds stably and specifically to CD4 T cells recognizing the HA110-120 peptide. Doxorubicin, a powerful antimitogenic anthracycline, was enzymatically assembled on the galactose residues of a DEF chimera. The DEF-gal-Dox construct preserved both the binding capacity to hemagglutinin (HA)-specific T cells, and the drug toxicity. Brief exposure of HA-specific T cells to DEF-gal-Dox construct in vitro was followed by drug internalization in the lysosomes, translocation to the nucleus, and apoptosis. Administration of DEF-gal-Dox to mice expressing the TCR-HA transgene reduced the frequency of TCR-HA T cells in the spleen and thymus by 27% and 42%, and inhibited HA proliferative capacity by 40% and 60%, respectively. It has not been demonstrated previously that pharmacologically active drugs able to modulate T-cell functions can be delivered to T cells in an antigen-specific manner by soluble, bivalent MHC II–peptide chimeras.

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Figure 1: Schematic representation of DEF-gal-Dox synthesis.
Figure 2: Specificity of doxorubicin coupling to the galactose residues of DEF chimera.
Figure 3: Delivery of doxorubicin by DEF-gal-Dox in the lysosomal compartment of antigen-specific T cells.
Figure 4: Determination of doxorubicin bound to DNA in T cells treated with DEF-gal-Dox construct.
Figure 5: Inhibition of HA-specific proliferation of TCR-HA T cells on exposure to DEF-gal-Dox in vitro.
Figure 6: Inhibition of HA-specific proliferation of TCR-HA T cells by DEF-gal-Dox in vivo.

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Acknowledgements

This work was supported by grants to S.C. (JDIF 1-1999-272, A & A .L. Sinsheimer Foundation, and NIH/ORWH/NIDDK 1R55-DK55744), and to T-D. B. (GC0 0247-3631/1999 from Mount Sinai School of Medicine, New York, NY, and NIH/NIDDK 1R41-DK55461-01A1).

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Authors and Affiliations

  1. Department of Microbiology, Mount Sinai School of Medicine, New York, 10029, NY

    Sofia Casares, Constantin A. Bona & Teodor- D. Brumeanu

  2. Institute of Neuropathology, Hannover Medical School, Hannover, 30625, Germany

    Alexandru C. Stan

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  1. Sofia Casares
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Correspondence to Teodor- D. Brumeanu.

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Casares, S., Stan, A., Bona, C. et al. Antigen-specific downregulation of T cells by doxorubicin delivered through a recombinant MHC II–peptide chimera. Nat Biotechnol 19, 142–147 (2001). https://doi.org/10.1038/84404

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