title: Silencing human NKG2D, DAP10, and DAP12 reduces cytotoxicity of activated CD8+ T cells and NK cells.
abstract: Human CD8+ T cells activated and expanded by TCR cross-linking and high-dose IL-2 acquire potent cytolytic ability against tumors and are a promising approach for immunotherapy of malignant diseases. We have recently reported that in vitro killing by these activated cells, which share phenotypic and functional characteristics with NK cells, is mediated principally by NKG2D. NKG2D is a surface receptor that is expressed by all NK cells and transmits an activating signal via the DAP10 adaptor molecule. Using stable RNA interference induced by lentiviral transduction, we show that NKG2D is required for cytolysis of tumor cells, including autologous tumor cells from patients with ovarian cancer. We also demonstrated that NKG2D is required for in vivo antitumor activity. Furthermore, both activated and expanded CD8+ T cells and NK cells use DAP10. In addition, direct killing was partially dependent on the DAP12 signaling pathway. This requirement by activated and expanded CD8+ T cells for DAP12, and hence stimulus from a putative DAP12-partnered activating surface receptor, persisted when assayed by anti-NKG2D Ab-mediated redirected cytolysis. These studies demonstrated the importance of NKG2D, DAP10, and DAP12 in human effector cell function.
authors.affiliation: Division of Blood and Marrow Transplantation, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Mobin
authors.initials: M
authors.lastname: Karimi
authors.forename: Thai M
authors.initials: TM
authors.lastname: Cao
authors.forename: Jeanette A
authors.initials: JA
authors.lastname: Baker
authors.forename: Michael R
authors.initials: MR
authors.lastname: Verneris
authors.forename: Luis
authors.initials: L
authors.lastname: Soares
authors.forename: Robert S
authors.initials: RS
authors.lastname: Negrin
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D005260:Female; D006801:Humans; D016219:Immunotherapy, Adoptive; D007694:Killer Cells, Natural / Q000276:immunology*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D016513:Mice, SCID; D055655:NK Cell Lectin-Like Receptor Subfamily K; D009374:Neoplasms, Experimental / Q000473:pathology / Q000628:therapy; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D011971:Receptors, Immunologic / Q000235:genetics*; D055607:Receptors, Natural Killer Cell; D014161:Transduction, Genetic
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C120528:HCST protein, human; C527085:KLRK1 protein, human; C527086:Klrk1 protein, mouse; D008565:Membrane Proteins; D055655:NK Cell Lectin-Like Receptor Subfamily K; D034741:RNA, Small Interfering; D011971:Receptors, Immunologic; D055607:Receptors, Natural Killer Cell; C110546:TYROBP protein, human