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Sustained IL-2R signaling of limited duration by high-dose mIL-2/mCD25 fusion protein amplifies tumor-reactive CD8+ T cells to enhance antitumor immunity

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Abstract

High-dose IL-2 induces cancer regression but its therapeutic use is limited due to high toxicities resulting from its broad cell targeting. In one strategy to overcome this limitation, IL-2 has been modified to selectively target the intermediate affinity IL-2R that broadly activates memory-phenotypic CD8+ T and NK cells, while minimizing Treg-associated tolerance. In this study, we modeled an alternative strategy to amplify tumor antigen-specific TCR transgenic CD8+ T cells through limited application of a long-acting IL-2 fusion protein, mIL-2/mCD25, which selectively targets the high-affinity IL-2R. Here, mice were vaccinated with a tumor antigen and high-dose mIL-2/mCD25 was applied to coincide with the induction of the high affinity IL-2R on tumor-specific T cells. A single high dose of mIL-2/mCD25, but not an equivalent amount of IL-2, amplified the frequency and function of tumor-reactive CD8+ T effector (Teff) and memory cells. These mIL-2/mCD25-dependent effects relied on distinctive requirements for TLR signals during priming of CD8+ tumor-specific T cells. The mIL-2/mCD25-amplified tumor-reactive effector and memory T cells supported long-lasting antitumor responses to B16-F10 melanoma. This regimen only transiently increased Tregs, yielding a favorable Teff–Treg ratio within the tumor microenvironment. Notably, mIL-2/mCD25 did not increase non-tumor-specific Teff or NK cells within tumors, further substantiating the specificity of mIL-2/mCD25 for tumor antigen-activated T cells. Thus, the selectivity and persistence of mIL-2/mCD25 in conjunction with a tumor vaccine supports antitumor immunity through a mechanism that is distinct from recombinant IL-2 or IL-2-based biologics that target the intermediate affinity IL-2R.

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Availability of data and materials

Data are available upon reasonable request. All data relevant to the study are included in the article or uploaded as supplementary information.

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Acknowledgements

We thank Michael Dee and Aixin Yu for technical assistance, Mary Struthers and Francisco Ramirez-Valle at Bristol Myers Squibb for critically reading the manuscript, and Patricia Guevara, Jay Enten, and Shannon Saigh from the Flow Cytometry Core of the Sylvester Comprehensive Cancer Center (supported by NIH P30CA240139).

Funding

This research was supported by funding to T.R.M. from the NIH (R21CA195334) and Sylvester Comprehensive Cancer Center at the University of Miami.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA

    Rosmely Hernandez, Kevin H. Toomer, Janika Põder, Alicia Santos Savio, Sunnie Hsiung & Thomas R. Malek

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  1. Rosmely Hernandez
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  2. Kevin H. Toomer
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Contributions

Conception and design: RH and TRM. Production and validation of mIL-2/mCD25: ASS. Acquisition of data: RH, KHT, JP, SH. Analysis and interpretation of data: RH and TRM. Manuscript Writing: RH and TRM. Revisions: All authors edited and approved the manuscript.

Corresponding author

Correspondence to Thomas R. Malek.

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Conflict of interest

The University of Miami, T.R.M. and R.H. have patents pending on IL-2/CD25 fusion proteins (Wo2016022671A1;T.R.M) and their use (PCT/US20/13152; T.R.M., R.H) that have been licensed exclusively to Bristol Myers Squibb, and some research on IL-2/CD25 fusion proteins has been supported in part by a collaboration and sponsored research and licensing agreement with Bristol Myers Squibb. The other authors have no financial conflicts of interest.

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Animal studies were approved by the Institutional Animal Care and Use Committee at the University of Miami (Protocol 18-147).

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Hernandez, R., Toomer, K.H., Põder, J. et al. Sustained IL-2R signaling of limited duration by high-dose mIL-2/mCD25 fusion protein amplifies tumor-reactive CD8+ T cells to enhance antitumor immunity. Cancer Immunol Immunother 70, 909–921 (2021). https://doi.org/10.1007/s00262-020-02722-5

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