Design and characterisation of a novel interleukin-15 receptor alpha fusion protein and analysis of interleukin-15 complexation

PLoS One. 2019 Jul 26;14(7):e0219313. doi: 10.1371/journal.pone.0219313. eCollection 2019.

Abstract

Interleukin-15 (IL15) is one of the most important cytokines currently being considered for cancer therapy applications. It is thought that by administering IL15 in complex with its cognate receptor alpha chain (IL15Rα) its biological activity could be increased manifold. We produced a fusion protein of mouse IL15Rα and the F8 antibody, that targets the alternatively-spliced extra-domain A (EDA) of fibronectin, which is overexpressed in many types of cancer. The fusion protein F8IL15Rα was cloned, expressed and characterized in vitro and its ability to bind to mouse IL15 was assessed with both size exclusion chromatography (SEC) and surface plasmon resonance (SPR) experiments. Furthermore, mouse and human IL15 and their corresponding Fc fused IL15Rα subunits were purchased, characterized and used to compare the capacity of F8IL15Rα to generate complexes. Surprisingly, none of the IL15Rα fusion proteins showed IL15 complexation on SEC. However, on SPR, F8IL15Rα displayed the ability to bind IL15. In a cell-based activity assay none of the IL15Rα fusions were able to increase cellular proliferation in combination with IL15 compared to IL15 alone. A better understanding of the molecular requirements for effective IL15 signalling are likely to be important for the development of IL15-based biopharmaceuticals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Interleukin-15 / metabolism*
  • Interleukin-15 Receptor alpha Subunit / metabolism*
  • Mice
  • Recombinant Fusion Proteins / metabolism*

Substances

  • Interleukin-15
  • Interleukin-15 Receptor alpha Subunit
  • Recombinant Fusion Proteins

Grants and funding

D.N. is grateful to ETH Zürich, the Swiss National Science Foundation [Project Nr. 310030B_163479/1] and the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program [grant agreement No 607603] for the financial support. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.