Specific engagement of the CD94/NKG2-A killer inhibitory receptor by the HLA-E class Ib molecule induces SHP-1 phosphatase recruitment to tyrosine-phosphorylated NKG2-A: evidence for receptor function in heterologous transfectants

Eur J Immunol. 1998 Apr;28(4):1280-91. doi: 10.1002/(SICI)1521-4141(199804)28:04<1280::AID-IMMU1280>3.0.CO;2-O.

Abstract

It has been recently demonstrated that the CD94/NKG2-A killer inhibitory receptor (KIR) specifically recognizes the HLA-E class Ib molecule. Moreover, the apparent CD94-mediated specific recognition of different HLA class Ia allotypes, transfected into the HLA-defective cell line 721.221, indeed depends on their selective ability to concomitantly stabilize the surface expression of endogenous HLA-E molecules, which confer protection against CD94/NKG2-A+ effector cells. In the present study, we show that a selective engagement of the CD94/NKG2-A inhibitory receptor with a specific monoclonal antibody (mAb) (Z199) was sufficient to induce tyrosine phosphorylation of the NKG2-A subunit and SHP-1 recruitment. These early biochemical events, commonly related to negative signaling pathways, were also detected upon the specific interaction of NK cells with an HLA-E+ 721.221 transfectant (.221-AEH), and were prevented by pre-incubation of .221-AEH with an anti-HLA class I mAb. Furthermore, mAb cross-linking of the CD94/NKG2-A receptor, segregated from other NK-associated molecules by transfection into a rat basophilic leukemia cell line (RBL-2H3), promoted tyrosine phosphorylation of NKG2-A and co-precipitation of SHP-1, together with an inhibition of secretory events triggered via Fc epsilonRI. Remarkably, interaction of CD94/NKG2-A+ RBL cells with the HLA-E+ .221-AEH transfectant specifically induced a detectable association of SHP-1 with NKG2-A, constituting a more formal evidence for the receptor-HLA class I interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / immunology*
  • Cell Line
  • HLA Antigens / immunology*
  • Histocompatibility Antigens Class I / immunology*
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Killer Cells, Natural / immunology*
  • Lectins, C-Type*
  • Ligands
  • Membrane Glycoproteins / immunology*
  • NK Cell Lectin-Like Receptor Subfamily D
  • Phosphorylation
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6
  • Protein Tyrosine Phosphatases / immunology*
  • Rats
  • Signal Transduction / immunology*
  • Transfection
  • Tyrosine

Substances

  • Antigens, CD
  • HLA Antigens
  • HLA-E antigen
  • Histocompatibility Antigens Class I
  • Intracellular Signaling Peptides and Proteins
  • KLRD1 protein, human
  • Klrd1 protein, rat
  • Lectins, C-Type
  • Ligands
  • Membrane Glycoproteins
  • NK Cell Lectin-Like Receptor Subfamily D
  • Tyrosine
  • PTPN11 protein, human
  • PTPN6 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6
  • Protein Tyrosine Phosphatases
  • Ptpn11 protein, rat
  • Ptpn6 protein, rat