title: Respiratory syncytial virus glycoprotein G interacts with DC-SIGN and L-SIGN to activate ERK1 and ERK2.
abstract: Respiratory syncytial virus (RSV) interaction with epithelial and dendritic cells (DCs) is known to require divalent cations, suggesting involvement of C-type lectins. RSV infection and maturation of primary human DCs are reduced in a dose-dependent manner by EDTA. Therefore, we asked whether RSV infection involves DC-SIGN (CD209) or its isoform L-SIGN (CD299) (DC-SIGN/R). Using surface plasmon resonance analysis, we demonstrated that the attachment G glycoprotein of RSV binds both DC- and L-SIGN. However, neutralization of DC- and L-SIGN on primary human DCs did not inhibit RSV infection, demonstrating that interactions between RSV G and DC- or L-SIGN are not required for productive infection. Thus, neither DC- nor L-SIGN represents a functional receptor for RSV. However, inhibition of these interactions increased DC activation, as evidenced by significantly higher levels of alpha interferon (IFN-α), MIP-1α, and MIP-1β in plasmacytoid DCs (pDCs) exposed to RSV after neutralization of DC-and L-SIGN. To understand the molecular interactions involved, intracellular signaling events triggered by purified RSV G glycoprotein were examined in DC- and L-SIGN-transfected 3T3 cells. RSV G interaction with DC- or L-SIGN was shown to stimulate ERK1 and ERK2 phosphorylation, with statistically significant increases relative to mock-infected cells. Neutralization of DC- and L-SIGN reduced ERK1/2 phosphorylation. With increased DC activation following DC- and L-SIGN neutralization and RSV exposure, these data demonstrate that the signaling events mediated by RSV G interactions with DC/L-SIGN are immunomodulatory and diminish DC activation, which may limit induction of RSV-specific immunity.
authors.affiliation: Viral Pathogenesis Laboratorya and Structural Biology Section,b Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA. tjohnson@genvec.com
authors.forename: Teresa R
authors.initials: TR
authors.lastname: Johnson
authors.forename: Jason S
authors.initials: JS
authors.lastname: McLellan
authors.forename: Barney S
authors.initials: BS
authors.lastname: Graham
mesh_terms: D002413:Cations, Divalent; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012136:Respiratory Syncytial Viruses / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000378:metabolism*; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D002413:Cations, Divalent; D015815:Cell Adhesion Molecules; D048049:Extracellular Signal-Regulated MAP Kinases; D037181:Lectins, C-Type; D011956:Receptors, Cell Surface; D014760:Viral Fusion Proteins; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C433029:CLEC4M protein, human; C502275:G glycoprotein, Respiratory syncytial virus
references.citation: J Mol Biol. 2005 Apr 15;347(5):979-89
references.pmid: 15784257
references.citation: Infect Immun. 1982 Mar;35(3):1142-6
references.pmid: 6175576
references.citation: J Immunol. 2009 Nov 1;183(9):5694-704
references.pmid: 19828628
references.citation: J Virol. 2003 Nov;77(21):11303-11
references.pmid: 14557616
references.citation: PLoS Pathog. 2007 Nov;3(11):e169
references.pmid: 17983270
references.citation: Future Microbiol. 2009 Sep;4(7):879-90
references.pmid: 19722841
references.citation: Ann Rheum Dis. 2008 Dec;67 Suppl 3:iii70-4
references.pmid: 19022819
references.citation: J Virol. 2008 Jun;82(12):5715-24
references.pmid: 18367533
references.citation: Cell Signal. 2010 Oct;22(10):1397-405
references.pmid: 20363321
references.citation: J Virol. 2005 May;79(9):5507-15
references.pmid: 15827165
references.citation: Virology. 2000 Jun 5;271(2):264-75
references.pmid: 10860881
references.citation: J Virol. 2010 May;84(9):4366-82
references.pmid: 20164223
references.citation: J Immunol. 2004 Nov 15;173(10):5935-43
references.pmid: 15528327
references.citation: Viral Immunol. 2000;13(1):125-35
references.pmid: 10733174
references.citation: J Virol. 2003 Feb;77(3):1747-56
references.pmid: 12525608
references.citation: J Virol. 2009 Oct;83(19):9682-93
references.pmid: 19625398
references.citation: Clin Exp Immunol. 2008 Feb;151(2):359-66
references.pmid: 18062796
references.citation: PLoS One. 2011 Jan 13;6(1):e15756
references.pmid: 21249196
references.citation: J Immunol. 2000 Nov 15;165(10):5839-46
references.pmid: 11067944
references.citation: J Virol. 2006 May;80(10):5032-40
references.pmid: 16641294
references.citation: J Virol. 2000 Jul;74(14):6442-7
references.pmid: 10864656
references.citation: Virology. 2009 Mar 30;386(1):192-202
references.pmid: 19185899
references.citation: J Biol Chem. 2005 Jun 24;280(25):23698-708
references.pmid: 15855154
references.citation: J Virol. 1994 Jul;68(7):4538-46
references.pmid: 8207828
references.citation: Am J Respir Cell Mol Biol. 2006 Mar;34(3):320-9
references.pmid: 16284360
references.citation: Clin Exp Allergy. 2010 Jan;40(1):111-22
references.pmid: 20205699
references.citation: Am J Respir Cell Mol Biol. 2004 Aug;31(2):193-9
references.pmid: 15016617
references.citation: Biol Chem. 2008 Oct;389(10):1273-82
references.pmid: 18713014
references.citation: Am J Respir Cell Mol Biol. 2000 Nov;23(5):586-92
references.pmid: 11062136
references.citation: PLoS One. 2011 Jan 28;6(1):e16458
references.pmid: 21297989
references.citation: J Exp Med. 2003 Jan 6;197(1):7-17
references.pmid: 12515809
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15748-53
references.pmid: 15496474
references.citation: Nat Rev Immunol. 2009 Jul;9(7):465-79
references.pmid: 19521399
references.citation: Nat Immunol. 2000 Nov;1(5):398-401
references.pmid: 11062499
references.citation: J Virol. 2005 Apr;79(8):4589-98
references.pmid: 15795245
references.citation: Nat Immunol. 2010 May;11(5):419-26
references.pmid: 20364151
references.citation: J Infect Dis. 1992 May;165(5):819-25
references.pmid: 1373752
references.citation: J Clin Microbiol. 1988 Jan;26(1):139-41
references.pmid: 3343306
references.citation: Clin Exp Immunol. 2006 Mar;143(3):513-22
references.pmid: 16487251
references.citation: Cell. 2000 Mar 3;100(5):587-97
references.pmid: 10721995
references.citation: Arch Virol. 2011 Jul;156(7):1275-9
references.pmid: 21424730
references.citation: J Clin Microbiol. 1986 Jun;23(6):1009-14
references.pmid: 3754878
references.citation: Biol Cell. 2010 Mar 25;102(6):335-50
references.pmid: 20356318
references.citation: Nat Immunol. 2009 Oct;10(10):1081-8
references.pmid: 19718030
references.citation: Immunity. 2009 Sep 18;31(3):412-24
references.pmid: 19766084
references.citation: J Exp Med. 2003 Apr 7;197(7):823-9
references.pmid: 12682107
references.citation: J Virol. 2006 Feb;80(3):1290-301
references.pmid: 16415006
references.citation: Nat Immunol. 2001 Aug;2(8):732-8
references.pmid: 11477410
references.citation: J Virol. 1999 Aug;73(8):6610-7
references.pmid: 10400758
references.citation: J Immunol. 1984 Oct;133(4):1925-30
references.pmid: 6236259
references.citation: J Immunol. 2005 Nov 1;175(9):5904-11
references.pmid: 16237083
references.citation: Proc Natl Acad Sci U S A. 2004 Sep 28;101(39):14067-72
references.pmid: 15371595
references.citation: J Virol. 2008 Mar;82(5):2040-55
references.pmid: 17928346
references.citation: Pediatr Res. 2002 Sep;52(3):363-7
references.pmid: 12193668
references.citation: Blood. 2006 May 15;107(10):3950-8
references.pmid: 16434485
references.citation: Nat Genet. 2005 May;37(5):507-13
references.pmid: 15838506
references.citation: Cell Biochem Biophys. 2010;56(1):49-58
references.pmid: 19851890
references.citation: J Virol. 2002 Jul;76(13):6841-4
references.pmid: 12050398
references.citation: J Infect Dis. 1999 Dec;180(6):2009-13
references.pmid: 10558959
references.citation: J Immunol. 2009 Nov 15;183(10):6588-99
references.pmid: 19846880
references.citation: J Infect Dis. 2002 Mar 1;185(5):682-5
references.pmid: 11865426