Structural Insights into the Inhibitory Mechanism of an Antibody against B7-H6, a Stress-Induced Cellular Ligand for the Natural Killer Cell Receptor NKp30

J Mol Biol. 2016 Nov 6;428(22):4457-4466. doi: 10.1016/j.jmb.2016.09.011. Epub 2016 Sep 20.


Antibodies have been shown to block signaling through cell surface receptors using several mechanisms. The two most common are binding to the ligand-binding site of the receptor and, conversely, binding to the receptor-binding site of the ligand. Here, we investigated the inhibitory mechanism of an antibody (17B1.3) against human B7-H6, a stress-induced cellular ligand for the natural killer (NK) cell receptor NKp30. Binding of this antibody to B7-H6, a transmembrane protein expressed on tumor and other stressed cells, but not on normal cells, prevents NK cell activation via NKp30. We determined the crystal structure of antibody 17B1.3 in complex with the ectodomain of B7-H6 to 2.5Å resolution. Surprisingly, 17B1.3 binds to a site on B7-H6 that is completely distinct from the binding site for NKp30, such that 17B1.3 does not block the NKp30-B7-H6 interaction. We then asked whether 17B1.3 prevents signaling by binding to a putative site for B7-H6 dimerization. However, structure-based mutations designed to disrupt potential B7-H6 dimerization through this site did not diminish NKp30-mediated cell activation. We conclude that the bulky 17B1.3 antibody most likely acts by sterically interfering with close cell-cell contacts at the NK cell-target cell interface that are required for NK cell activation. A similar inhibitory mechanism may apply to other antibodies, including therapeutic antibodies that block signaling through cell surface receptors whose ligands are also cell surface proteins.

Keywords: NK cell activation; NK cell receptor; antibody; antibody–ligand complex; crystal structure.

Publication types

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

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / metabolism*
  • B7 Antigens / antagonists & inhibitors*
  • B7 Antigens / chemistry
  • B7 Antigens / genetics
  • B7 Antigens / metabolism*
  • Crystallography, X-Ray
  • DNA Mutational Analysis
  • Humans
  • Models, Molecular
  • Natural Cytotoxicity Triggering Receptor 3 / metabolism*
  • Protein Conformation


  • Antibodies, Monoclonal
  • B7 Antigens
  • NCR3 protein, human
  • NCR3LG1 protein, human
  • Natural Cytotoxicity Triggering Receptor 3