Ab-IL2 fusion proteins mediate NK cell immune synapse formation by polarizing CD25 to the target cell-effector cell interface

Cancer Immunol Immunother. 2011 Dec;60(12):1789-800. doi: 10.1007/s00262-011-1072-9. Epub 2011 Jul 27.

Abstract

The huKS-IL2 immunocytokine (IC) consists of IL2 fused to a mAb against EpCAM, while the hu14.18-IL2 IC recognizes the GD2 disialoganglioside. They are under evaluation for treatment of EpCAM(+) (ovarian) and GD2(+) (neuroblastoma and melanoma) malignancies because of their proven ability to enhance tumor cell killing by antibody-dependent cell-mediated cytotoxicity (ADCC) and by antitumor cytotoxic T cells. Here, we demonstrate that huKS-IL2 and hu14.18-IL2 bind to tumor cells via their antibody components and increase adhesion and activating immune synapse (AIS) formation with NK cells by engaging the immune cells' IL-2 receptors (IL2R). The NK leukemia cell line, NKL (which expresses high affinity IL2Rs), shows fivefold increase in binding to tumor targets when treated with IC compared to matching controls. This increase in binding is effectively inhibited by blocking antibodies against CD25, the α-chain of the IL2R. NK cells isolated from the peritoneal environment of ovarian cancer patients, known to be impaired in mediating ADCC, bind to huKS-IL2 via CD25. The increased binding between tumor and effector cells via ICs is due to the formation of AIS that are characterized by the simultaneous polarization of LFA-1, CD2 and F-actin at the cellular interface. AIS formation of peritoneal NK and NKL cells is inhibited by anti-CD25 blocking antibody and is 50-200% higher with IC versus the parent antibody. These findings demonstrate that the IL-2 component of the IC allows IL2Rs to function not only as receptors for this cytokine but also as facilitators of peritoneal NK cell binding to IC-coated tumor cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies, Monoclonal
  • Cell Line, Tumor
  • Cell Separation
  • Female
  • Flow Cytometry
  • Gangliosides / immunology
  • Humans
  • Immunological Synapses / immunology*
  • Immunotherapy / methods*
  • Interleukin-2 / analogs & derivatives*
  • Interleukin-2 / immunology
  • Interleukin-2 Receptor alpha Subunit / immunology*
  • Interleukin-2 Receptor alpha Subunit / metabolism
  • Killer Cells, Natural / immunology*
  • Lymphocyte Activation / immunology
  • Microscopy, Confocal
  • Ovarian Neoplasms / immunology
  • Protein Transport / immunology
  • Recombinant Fusion Proteins

Substances

  • Antibodies, Monoclonal
  • Gangliosides
  • Interleukin-2
  • Interleukin-2 Receptor alpha Subunit
  • Recombinant Fusion Proteins
  • tucotuzumab celmoleukin
  • ganglioside, GD2