Design and characterization of a protein superagonist of IL-15 fused with IL-15Rα and a high-affinity T cell receptor

Jennifer D. Stone, Adam S. Chervin, Hans Schreiber, David M. Kranz

Research output: Contribution to journalArticlepeer-review

Abstract

To avoid high systemic doses, strategies involving antigen-specific delivery of cytokine via linked antibodies or antibody fragments have been used. Targeting cancer-associated peptides presented by major histocompatibility complex (MHC) molecules (pepMHC) increases the number of potential target antigens and takes advantage of cross-presentation on tumor stroma and in draining lymph nodes. Here, we use a soluble, high-affinity single-chain T cell receptor Vα-Vβ (scTv), to deliver cytokines to intracellular tumor-associated antigens presented as pepMHC. As typical wild-type T cell receptors (TCRs) exhibit low affinity (Kd = 1-100 μM or more), we used an engineered TCR, m33, that binds its antigenic peptide SIYRYYGL (SIY) bound to the murine class I major histocompatability complex protein H2-Kb (SIY/Kb) with nanomolar affinity (Kd = 30 nM). We generated constructs consisting of m33 scTv fused to murine interleukin 2 (IL-2), interleukin 15 (IL-15), or IL-15/IL-15Rα (IL-15 linked to IL-15Rα sushi domain, called "superfusion"). The fusions were purified with good yields and bound specifically to SIY/Kb with high affinity. Proper cytokine folding and binding were confirmed, and the fusions were capable of stimulating proliferation of cytokine-dependent cells, both when added directly and when presented in trans, bound to cells with the target pepMHC. The m33 superfusion was particularly potent and stable and represents a promising design for targeted antitumor immunomodulation.

Original languageEnglish (US)
Pages (from-to)1588-1597
Number of pages10
JournalBiotechnology Progress
Volume28
Issue number6
DOIs
StatePublished - Nov 2012

Keywords

  • High-affinity TCR
  • IL-15/IL-15Rα
  • IL-2
  • Immunokine
  • TCR-cytokine fusion

ASJC Scopus subject areas

  • Biotechnology

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