title: Rapid activation receptor- or IL-2-induced lytic granule convergence in human natural killer cells requires Src, but not downstream signaling.
abstract: Natural killer (NK) cells participate in host defense by surveying for and ultimately killing virally infected or malignant target cells. NK cell cytotoxicity is a tightly regulated process that proceeds stepwise from adhesion and activation to the secretion of preformed lytic granule contents onto a diseased or stressed cell. We previously characterized rapid dynein-dependent lytic granule convergence to the microtubule-organizing center (MTOC) as an early, prerequisite step in NK cell cytotoxicity. Although multiple activating receptors can trigger granule convergence, the specific signal or signals responsible remained unknown. Using live cell confocal microscopy, NK cell lytic granule movement after NK cell activation was captured and measured. Using inhibitors of common early signaling mediators, we show that Src kinases are required for lytic granule convergence, but downstream signals that promote actin rearrangement, MTOC polarization, and calcium mobilization are not. Exposure to interleukin 2 was also sufficient to induce lytic granule convergence, which required noncanonical Src-dependent signaling. Thus, NK cell lytic granule convergence, prompted by specific receptor-mediated and soluble cytokine signals, depends on a directly downstream early Src kinase-dependent signal and emphasizes the importance of this step in readying NK cells for cytotoxicity.
authors: Graduate Program in Cell Biology and Physiology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
authors: Ashley Mentlik
authors: AM
authors: James
authors: Hsiang-Ting
authors: HT
authors: Hsu
authors: Prachi
authors: P
authors: Dongre
authors: Gulbu
authors: G
authors: Uzel
authors: Emily M
authors: EM
authors: Mace
authors: Pinaki P
authors: PP
authors: Banerjee
authors: Jordan S
authors: JS
authors: Orange
mesh_terms: D000199:Actins / Q000378:metabolism; D018821:CD18 Antigens / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000276:immunology / Q000378:metabolism*; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004398:Dyneins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007376:Interleukin-2 / Q000276:immunology / Q000378:metabolism*; D053616:Janus Kinase 3 / Q000378:metabolism; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D008213:Lymphocyte Activation / Q000276:immunology*; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism; D018613:Microscopy, Confocal; D022101:Microtubule-Organizing Center / Q000276:immunology / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051966:Phospholipase C gamma / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D015398:Signal Transduction / Q000276:immunology*; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000199:Actins; D018821:CD18 Antigens; D007376:Interleukin-2; D047428:Protein Kinase Inhibitors; D049452:Green Fluorescent Proteins; D019869:Phosphatidylinositol 3-Kinases; C507933:JAK3 protein, human; D053616:Janus Kinase 3; D019061:src-Family Kinases; D020930:MAP Kinase Kinase Kinases; D051966:Phospholipase C gamma; D004398:Dyneins
references: J Biol Chem. 2004 Apr 16;279(16):16194-205
references: 14960575
references: Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15062-7
references: 10611338
references: J Clin Invest. 2011 Apr;121(4):1535-48
references: 21383498
references: J Biol Chem. 1994 Jan 21;269(3):1599-602
references: 8294403
references: Nat Immunol. 2000 Nov;1(5):419-25
references: 11062502
references: J Biol Chem. 1998 Aug 14;273(33):21194-202
references: 9694876
references: J Exp Med. 1996 Dec 1;184(6):2243-50
references: 8976179
references: Blood. 2003 Apr 15;101(8):3052-7
references: 12480696
references: J Immunol. 2003 Jun 15;170(12):6107-14
references: 12794140
references: Mol Cell Biol. 2003 Sep;23(17):6291-9
references: 12917349
references: Mol Biol Cell. 1997 Sep;8(9):1709-21
references: 9307968
references: PLoS One. 2008 Sep 26;3(9):e3278
references: 18818767
references: Mol Biol Cell. 2010 Jul 1;21(13):2241-56
references: 20444980
references: Nat Immunol. 2003 Jun;4(6):557-64
references: 12740575
references: J Clin Invest. 2012 Oct;122(10):3769-80
references: 23006327
references: J Immunol. 1994 Mar 1;152(5):2098-104
references: 7907631
references: Nat Rev Immunol. 2008 Sep;8(9):713-25
references: 19172692
references: Blood. 2006 Jan 1;107(1):159-66
references: 16150947
references: J Immunol. 2006 Aug 15;177(4):2349-55
references: 16887996
references: Proc Natl Acad Sci U S A. 2008 Feb 26;105(8):3017-22
references: 18287025
references: J Exp Med. 2003 Aug 4;198(3):469-74
references: 12885870
references: J Immunol. 2011 Mar 1;186(5):2998-3005
references: 21270398
references: Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3725-9
references: 8097322
references: J Biol Chem. 1999 Sep 24;274(39):27583-9
references: 10488096
references: J Immunol. 2005 Jul 1;175(1):213-8
references: 15972651
references: J Immunol. 1999 Feb 15;162(4):2081-6
references: 9973481
references: Cell Immunol. 1993 Feb;146(2):391-405
references: 8174177
references: Immunity. 2011 Jun 24;34(6):905-18
references: 21703542
references: Proc Natl Acad Sci U S A. 2006 Jul 5;103(27):10346-10351
references: 16801532
references: J Immunol. 1984 Dec;133(6):2972-8
references: 6092461
references: J Biol Chem. 1996 Dec 20;271(51):33141-7
references: 8955163
references: Nat Immunol. 2006 May;7(5):524-32
references: 16582911
references: Curr Protoc Immunol. 2010 Aug;Chapter 11:Unit 11.9B
references: 20814939
references: Proc Natl Acad Sci U S A. 2007 Apr 10;104(15):6329-34
references: 17395718
references: J Biol Chem. 2000 Jul 21;275(29):21946-52
references: 10781580
references: J Immunol. 2006 Jun 1;176(11):6615-23
references: 16709819
references: J Virol. 2011 May;85(9):4623-7
references: 21345943
references: Annu Rev Med. 1987;38:175-94
references: 3555290
references: J Immunol. 1991 Oct 1;147(7):2202-7
references: 1655887
references: Eur J Immunol. 2003 May;33(5):1235-41
references: 12731048
references: Nat Rev Immunol. 2011 Aug 26;11(10):645-57
references: 21869816
references: J Exp Med. 2007 Oct 1;204(10):2305-20
references: 17785506
references: PLoS Biol. 2011 Sep;9(9):e1001151
references: 21931536
references: Angiogenesis. 2007;10(3):167-82
references: 17486419
references: FASEB J. 1999 Jan;13(1):31-8
references: 9872927
references: J Leukoc Biol. 1996 Oct;60(4):441-52
references: 8864127