Signaling through human killer cell activating receptors triggers tyrosine phosphorylation of an associated protein complex

Our understanding of the biology of human natural killer (NK) cells has significantly advanced in recent years upon identification of a family of NK cell‐expressed genes that encode killer cell inhibitory receptors (KIR). Individual KIR can selectively bind various HLA class I allotypes and consequently transduce inhibitory signals that block NK cell lysis of ligand‐bearing target cells. A distinct subset of related and linked genes express truncated versions of KIR that are otherwise highly homologous in amino acid sequence. Interestingly, these receptors appear to transmit stimulatory signals into NK cells and have been termed killer cell activating receptors (KAR). In this report, we demonstrate that recognition of HLA‐Cw3 by the p50 KAR, NKAT8, can potentiate the cytotoxic response of appropriate NK cell clones. Specific cross‐linking of this KAR with a monoclonal antibody resulted in intracellular calcium mobilization, protein tyrosine phosphorylation, and phosphorylation of the MAP kinases, ERK1 and ERK2. In addition, we identified a KAR‐associated disulfide‐linked dimer of a 13‐kDa protein that was absent in the Jurkat T cell line and is predicted to participate in these activation signaling events. Upon treatment of NK cells with pervanadate, the disulfide‐linked p13 and additional proteins of 25, 30, 37 and 50 – 95 kDa were identified as KAR‐associated tyrosine phosphoproteins. Importantly, p13 was inducibly tyrosine phosphorylated upon cross‐linking of NKAT8, which strongly suggests that the associated p13 provides KAR with appropriate cytoplasmic structure to couple with tyrosine kinase‐mediated signaling effectors.

[1]  L. Lanier,et al.  CD94/NKG2 is the predominant inhibitory receptor involved in recognition of HLA-G by decidual and peripheral blood NK cells. , 1997, Journal of immunology.

[2]  K. Campbell,et al.  Normal development but differentially altered proliferative responses of lymphocytes in mice lacking CD81 , 1997, The EMBO journal.

[3]  H. Moriya,et al.  A new 12-kilodalton dimer associated with pre-TCR complex and clonotype-independent CD3 complex on immature thymocytes. , 1997, Journal of immunology.

[4]  P. Leibson Signal transduction during natural killer cell activation: inside the mind of a killer. , 1997, Immunity.

[5]  É. Vivier,et al.  Human killer cell activatory receptors for MHC class I molecules are included in a multimeric complex expressed by natural killer cells. , 1997, Journal of immunology.

[6]  L. Lanier,et al.  Natural killer cell cytolytic activity is inhibited by NKG2-A and activated by NKG2-C. , 1997, Journal of immunology.

[7]  L. Lanier Natural killer cells: from no receptors to too many. , 1997, Immunity.

[8]  W. Seaman,et al.  Mouse Ly-49A Interrupts Early Signaling Events in Natural Killer Cell Cytotoxicity and Functionally Associates with the SHP-1 Tyrosine Phosphatase , 1997, The Journal of experimental medicine.

[9]  C. Biron Activation and function of natural killer cell responses during viral infections. , 1997, Current opinion in immunology.

[10]  T. George,et al.  Role of murine NK cells and their receptors in hybrid resistance. , 1997, Current opinion in immunology.

[11]  M. Llano,et al.  Structure and function of the CD94 C‐type lectin receptor complex involved in recognition of HLA class I molecules , 1997, Immunological reviews.

[12]  M. Colonna Specificity and function of immunoglobulin superfamily NK cell inhibitory and stimulatory receptors , 1997, Immunological reviews.

[13]  O. Mandelboim,et al.  Enhancement of Class II-Restricted T cell Responses by Costimulatory NK Receptors for Class I MHC Proteins , 1996, Science.

[14]  I. Melero,et al.  Biochemical and serologic evidence for the existence of functionally distinct forms of the CD94 NK cell receptor. , 1996, Journal of immunology.

[15]  J. Kinet,et al.  Sequential involvement of Lck and SHP-1 with MHC-recognizing receptors on NK cells inhibits FcR-initiated tyrosine kinase activation. , 1996, Immunity.

[16]  S. Anderson,et al.  The Ly-49D Receptor Activates Murine Natural Killer Cells , 1996, The Journal of experimental medicine.

[17]  M. López-Botet,et al.  Clonotypic differences in signaling from CD94 (kp43) on NK cells lead to divergent cellular responses. , 1996, Journal of immunology.

[18]  R. Biassoni,et al.  A novel surface molecule homologous to the p58/p50 family of receptors is selectively expressed on a subset of human natural killer cells and induces both triggering of cell functions and proliferation , 1996, European journal of immunology.

[19]  M. Colonna,et al.  Tyrosine phosphorylation of a human killer inhibitory receptor recruits protein tyrosine phosphatase 1C , 1996, The Journal of experimental medicine.

[20]  L. Lanier,et al.  Phosphotyrosines in the killer cell inhibitory receptor motif of NKB1 are required for negative signaling and for association with protein tyrosine phosphatase 1C , 1996, The Journal of experimental medicine.

[21]  D. Marguet,et al.  Human and mouse killer-cell inhibitory receptors recruit PTP1C and PTP1D protein tyrosine phosphatases. , 1996, Journal of immunology.

[22]  M. Colonna,et al.  A human killer inhibitory receptor specific for HLA-A1,2. , 1996, Journal of immunology.

[23]  S. Sforzini,et al.  The natural killer-related receptor for HLA-C expressed on T cells from CD3+ lymphoproliferative disease of granular lymphocytes displays either inhibitory or stimulatory function. , 1996, Blood.

[24]  J. Ritz,et al.  Characterization of a cell line, NKL, derived from an aggressive human natural killer cell leukemia. , 1996, Experimental hematology.

[25]  R. Biassoni,et al.  The human leukocyte antigen (HLA)-C-specific "activatory" or "inhibitory" natural killer cell receptors display highly homologous extracellular domains but differ in their transmembrane and intracytoplasmic portions , 1996, The Journal of experimental medicine.

[26]  T. Mcclanahan,et al.  Molecular cloning of NKB1. A natural killer cell receptor for HLA-B allotypes. , 1995, Journal of immunology.

[27]  L. Moretta,et al.  Existence of both inhibitory (p58) and activatory (p50) receptors for HLA-C molecules in human natural killer cells , 1995, The Journal of experimental medicine.

[28]  Eric O Long,et al.  Molecular clones of the p58 NK cell receptor reveal immunoglobulin-related molecules with diversity in both the extra- and intracellular domains. , 1995, Immunity.

[29]  M. Colonna,et al.  Cloning of immunoglobulin-superfamily members associated with HLA-C and HLA-B recognition by human natural killer cells. , 1995, Science.

[30]  E. Palmer,et al.  CART: a conserved antigen receptor transmembrane motif. , 1994, Seminars in immunology.

[31]  G. Ferrara,et al.  NK3-specific natural killer cells are selectively inhibited by Bw4- positive HLA alleles with isoleucine 80 , 1994, The Journal of experimental medicine.

[32]  E. Ciccone,et al.  The human natural killer cell receptor for major histocompatibility complex class I molecules. Surface modulation of p58 molecules and their linkage to CD3 ζ chain, FcϵRI γ chain and the p56lck kinase , 1994 .

[33]  M. Balboa,et al.  Tyrosine kinase-dependent activation of human NK cell functions upon stimulation through a 58-kDa surface antigen selectively expressed on discrete subsets of NK cells and T lymphocytes. , 1994, Journal of immunology.

[34]  G. Ferrara,et al.  HLA-C is the inhibitory ligand that determines dominant resistance to lysis by NK1- and NK2-specific natural killer cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[35]  P. Parham,et al.  Specificity of HLA class I antigen recognition by human NK clones: evidence for clonal heterogeneity, protection by self and non-self alleles, and influence of the target cell type , 1993, The Journal of experimental medicine.

[36]  J. O’Shea,et al.  Activation of human peripheral blood T lymphocytes by pharmacological induction of protein-tyrosine phosphorylation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[37]  N. Green The semiotics of charge , 1991, Nature.

[38]  J. Bonifacino,et al.  Membrane protein association by potential intrarnembrane charge pairs , 1991, Nature.

[39]  J. Shabanowitz,et al.  Identification of the regulatory phosphorylation sites in pp42/mitogen‐activated protein kinase (MAP kinase). , 1991, The EMBO journal.

[40]  K. S. Campbell,et al.  B lymphocyte antigen receptors (mIg) are non‐covalently associated with a disulfide linked, inducibly phosphorylated glycoprotein complex. , 1990, The EMBO journal.

[41]  Arthur Weiss,et al.  The role of T3 surface molecules in the activation of human T cells: a two-stimulus requirement for IL 2 production reflects events occurring at a pre-translational level. , 1984, Journal of immunology.

[42]  P. Parham,et al.  Monomorphic anti-HLA-A,B,C monoclonal antibodies detecting molecular subunits and combinatorial determinants. , 1982, Journal of immunology.

[43]  R. Biassoni,et al.  Receptors for HLA class-I molecules in human natural killer cells. , 1996, Annual review of immunology.

[44]  Eric O Long,et al.  Recruitment of tyrosine phosphatase HCP by the killer cell inhibitor receptor. , 1996, Immunity.

[45]  H. Ljunggren,et al.  In search of the 'missing self': MHC molecules and NK cell recognition. , 1990, Immunology today.

[46]  E. Clark,et al.  Human T Lymphocyte Cell Surface Molecules Defined by the Workshop Monoclonal Antibodies (“T Cell Protocol”) , 1984 .

[47]  C. Heusser,et al.  Induction of resting B cells to DNA synthesis by soluble monoclonal anti‐immunoglobulin , 1984, European journal of immunology.