B Cell Repertoire Analysis Identifies New Antigenic Domains on Glycoprotein B of Human Cytomegalovirus which Are Target of Neutralizing Antibodies

English version České info

Human cytomegalovirus (HCMV), a herpesvirus, is a ubiquitously distributed pathogen that causes severe disease in immunosuppressed patients and infected newborns. Efforts are underway to prepare effective subunit vaccines and therapies including antiviral antibodies. However, current vaccine efforts are hampered by the lack of information on protective immune responses against HCMV. Characterizing the B-cell response in healthy infected individuals could aid in the design of optimal vaccines and therapeutic antibodies. To address this problem, we determined, for the first time, the B-cell repertoire against glycoprotein B (gB) of HCMV in different healthy HCMV seropositive individuals in an unbiased fashion. HCMV gB represents a dominant viral antigenic determinant for induction of neutralizing antibodies during infection and is also a component in several experimental HCMV vaccines currently being tested in humans. Our findings have revealed that the vast majority (>90%) of gB-specific antibodies secreted from B-cell clones do not have virus neutralizing activity. Most neutralizing antibodies were found to bind to epitopes not located within the previously characterized antigenic domains (AD) of gB. To map the target structures of these neutralizing antibodies, we generated a 3D model of HCMV gB and used it to identify surface exposed protein domains. Two protein domains were found to be targeted by the majority of neutralizing antibodies. Domain I, located between amino acids (aa) 133–343 of gB and domain II, a discontinuous domain, built from residues 121–132 and 344–438. Analysis of a larger panel of human sera from HCMV seropositive individuals revealed positivity rates of >50% against domain I and >90% against domain II, respectively. In accordance with previous nomenclature the domains were designated AD-4 (Dom II) and AD-5 (Dom I), respectively. Collectively, these data will contribute to optimal vaccine design and development of antibodies effective in passive immunization.

Vyšlo v časopise: B Cell Repertoire Analysis Identifies New Antigenic Domains on Glycoprotein B of Human Cytomegalovirus which Are Target of Neutralizing Antibodies. PLoS Pathog 7(8): e32767. doi:10.1371/journal.ppat.1002172
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002172

Souhrn

Zdroje

1. BoeckhMLjungmanP 2009 How we treat cytomegalovirus in hematopoietic cell transplant recipients. Blood 113 5711 5719

2. KennesonACannonMJ 2007 Review and meta-analysis of the epidemiology of congenital cytomegalovirus (CMV) infection. Rev Med Virol 17 253 276

3. LudwigAHengelH 2009 Epidemiological impact and disease burden of congenital cytomegalovirus infection in Europe. Euro Surveill 14 26 32

4. BoppanaSBPassRFBrittWJStagnoSAlfordCA 1992 Symptomatic congenital cytomegalovirus infection: neonatal morbidity and mortality. Pediatr Infect Dis J 11 93 99

5. DahleAJFowlerKBWrightJDBoppanaSBBrittWJ 2000 Longitudinal investigation of hearing disorders in children with congenital cytomegalovirus. J Am Acad Audiol 11 283 290

6. StrattonKRDurchJS 2001 Vaccines for the 21st century: a tool for decisionmaking. Washington, DC The National Academic Press

7. ZinkernagelRMHengartnerH 2006 Protective 'immunity' by pre-existent neutralizing antibody titers and preactivated T cells but not by so-called 'immunological memory'. Immunol Rev 211 310 319

8. NigroGAdlerSPLaTRBestAM 2005 Passive immunization during pregnancy for congenital cytomegalovirus infection. N Engl J Med 353 1350 1362

9. NigroGTorreRLPentimalliHTavernaPLituaniaM 2008 Regression of fetal cerebral abnormalities by primary cytomegalovirus infection following hyperimmunoglobulin therapy. Prenat Diagn 28 512 517

10. FowlerKBStagnoSPassRF 2003 Maternal immunity and prevention of congenital cytomegalovirus infection. JAMA 289 1008 1011

11. BurtonDR 2002 Antibodies, viruses and vaccines. Nat Rev Immunol 2 706 713

12. VarnumSMStreblowDNMonroeMESmithPAuberryKJ 2004 Identification of proteins in human cytomegalovirus (HCMV) particles: the HCMV proteome. J Virol 78 10960 10966

13. WangDShenkT 2005 Human cytomegalovirus virion protein complex required for epithelial and endothelial cell tropism. Proc Natl Acad Sci U S A 102 18153 18158

14. BrittWJVuglerLButfiloskiEJStephensEB 1990 Cell surface expression of human cytomegalovirus (HCMV) gp55-116 (gB): use of HCMV-recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. J Virol 64 1079 1085

15. ShimamuraMMachMBrittWJ 2006 Human cytomegalovirus infection elicits a glycoprotein M (gM)/gN-specific virus-neutralizing antibody response. J Virol 80 4591 4600

16. UrbanMKleinMBrittWJHassfurtherEMachM 1996 Glycoprotein H of human cytomegalovirus is a major antigen for the neutralizing humoral immune response. J Gen Virol 77 1537 1547

17. SchoppelKKropffBSchmidtCVornhagenRMachM 1997 The humoral immune response against human cytomegalovirus is characterized by a delayed synthesis of glycoprotein-specific antibodies. J Infect Dis 175 533 544

18. MarshallGSRabalaisGPStoutGGWaldeyerSL 1992 Antibodies to recombinant-derived glycoprotein B after natural human cytomegalovirus infection correlate with neutralizing activity. J Infect Dis 165 381 384

19. NavarroDPazPTugizovSToppKLa VailJPereiraL 1993 Glycoprotein B of human cytomegalovirus promotes virion penetration into cells, transmission of infection from cell to cell, and fusion of infected cells. Virology 197 143 158

20. SchleissMRBourneNBernsteinDI 2003 Preconception vaccination with a glycoprotein B (gB) DNA vaccine protects against cytomegalovirus (CMV) transmission in the guinea pig model of congenital CMV infection. J Infect Dis 188 1868 1874

21. CekinovicDGolemacMPugelEPTomacJCicin-SainL 2008 Passive immunization reduces murine cytomegalovirus-induced brain pathology in newborn mice. J Virol 82 12172 12180

22. AdlerSPPlotkinSAGonczolECadozMMericC 1999 A canarypox vector expressing cytomegalovirus (CMV) glycoprotein B primes for antibody responses to a live attenuated CMV vaccine (Towne). J Infect Dis 180 843 846

23. BernsteinDIReapEAKatenKWatsonASmithK 2009 Randomized, double-blind, Phase 1 trial of an alphavirus replicon vaccine for cytomegalovirus in CMV seronegative adult volunteers. Vaccine 28 484 493

24. PassRFZhangCEvansASimpsonTAndrewsW 2009 Vaccine prevention of maternal cytomegalovirus infection. N Engl J Med 360 1191 1199

25. FeireALKossHComptonT 2004 Cellular integrins function as entry receptors for human cytomegalovirus via a highly conserved disintegrin-like domain. Proc Natl Acad Sci U S A 101 15470 15475

26. SoroceanuLAkhavanACobbsCS 2008 Platelet-derived growth factor-alpha receptor activation is required for human cytomegalovirus infection. Nature 455 391 395

27. ComptonTNowlinDMCooperNR 1993 Initiation of human cytomegalovirus infection requires initial interaction with cell surface heparan sulfate. Virology 193 834 841

28. BoldSOhlinMGartenWRadsakK 1996 Structural domains involved in human cytomegalovirus glycoprotein B-mediated cell-cell fusion. J Gen Virol 77 Pt 9 2297 2302

29. VanarsdallALRyckmanBJChaseMCJohnsonDC 2008 Human cytomegalovirus glycoproteins gB and gH/gL mediate epithelial cell-cell fusion when expressed either in cis or in trans. J Virol 82 11837 11850

30. LopperMComptonT 2004 Coiled-coil domains in glycoproteins B and H are involved in human cytomegalovirus membrane fusion. J Virol 78 8333 8341

31. IsaacsonMKComptonT 2009 Human cytomegalovirus glycoprotein B is required for virus entry and cell-to-cell spread but not for virion attachment, assembly, or egress. J Virol 83 3891 3903

32. WagnerBKropffBKalbacherHBrittWSundqvistVA 1992 A continuous sequence of more than 70 amino acids is essential for antibody binding to the dominant antigenic site of glycoprotein gp58 of human cytomegalovirus. J Virol 66 5290 5297

33. SchoppelKHassfurtherEBrittWJOhlinMBorrebaeckCA 1996 Antibodies specific for the antigenic domain 1 (AD-1) of glycoprotein B (gpUL55) of human cytomegalovirus bind to different substructures. Virology 216 133 145

34. AulitzkyWESchulzTFTilgHNiederwieserDLarcherK 1991 Human monoclonal antibodies neutralizing cytomegalovirus (CMV) for prophylaxis of CMV disease: report of a phase I trial in bone marrow transplant recipients. J Infect Dis 163 1344 1347

35. OhlinMSundqvistVAMachMWahrenBBorrebaeckCA 1993 Fine specificity of the human immune response to the major neutralization epitopes expressed on cytomegalovirus gp58/116 (gB), as determined with human monoclonal antibodies. J Virol 67 703 710

36. SpecknerAGlykofrydesDOhlinMMachM 1999 Antigenic domain 1 of human cytomegalovirus glycoprotein B induces a multitude of different antibodies which, when combined, results in incomplete virus neutralization. J Gen Virol 80 2183 2191

37. MeyerHSundqvistVAPereiraLMachM 1992 Glycoprotein gp116 of human cytomegalovirus contains epitopes for strain-common and strain-specific antibodies. J Gen Virol 73 2375 2383

38. KniessNMachMFayJBrittWJ 1991 Distribution of linear antigenic sites on glycoprotein gp55 of human cytomegalovirus. J Virol 65 138 146

39. SilvestriMSundqvistVARudenUWahrenB 1991 Characterization of a major antigenic region on gp55 of human cytomegalovirus. J Gen Virol 72 3017 3023

40. NavarroDLennetteETugizovSPereiraL 1997 Humoral immune response to functional regions of human cytomegalovirus glycoprotein B. J Med Virol 52 451 459

41. MeyerHMasuhoYMachM 1990 The gp116 of the gp58/116 complex of human cytomegalovirus represents the amino-terminal part of the precursor molecule and contains a neutralizing epitope. J Gen Virol 71 2443 2450

42. PinnaDCortiDJarrossayDSallustoFLanzavecchiaA 2009 Clonal dissection of the human memory B-cell repertoire following infection and vaccination. Eur J Immunol 39 1260 1270

43. McLeanGROlsenOAWattINRathanaswamiPLeslieKB 2005 Recognition of human cytomegalovirus by human primary immunoglobulins identifies an innate foundation to an adaptive immune response. J Immunol 174 4768 4778

44. TraggiaiEBeckerSSubbaraoKKolesnikovaLUematsuY 2004 An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus. Nat Med 10 871 875

45. MachM 2005 Antibody-mediated neutralization of infectivity. Cytomegaloviruses: Molecular Biology and Immunology Caister Academic Press 265 283

46. HeldweinEELouHBenderFCCohenGHEisenbergRJHarrisonSC 2006 Crystal structure of glycoprotein B from herpes simplex virus 1. Science 313 217 220

47. HannahBPCairnsTMBenderFCWhitbeckJCLouHE 2009 Herpes simplex virus glycoprotein B associates with target membranes via its fusion loops. J Virol 83 6825 6836

48. LopperMComptonT 2002 Disulfide bond configuration of human cytomegalovirus glycoprotein B. J Virol 76 6073 6082

49. WangXHuongSMChiuMLRaab-TraubNHuangES 2003 Epidermal growth factor receptor is a cellular receptor for human cytomegalovirus. Nature 424 456 461

50. FeireALRoyRMManleyKComptonT 2010 The glycoprotein B disintegrin-like domain binds beta 1 integrin to mediate cytomegalovirus entry. J Virol 84 10026 10037

51. OhizumiYSuzukiHMatsumotoYMasuhoYNumazakiY 1992 Neutralizing mechanisms of two human monoclonal antibodies against human cytomegalovirus glycoprotein 130/55. J Gen Virol 73 2705 2707

52. ThrosbyMGeuijenCGoudsmitJBakkerAQKorimbocusJ 2006 Isolation and characterization of human monoclonal antibodies from individuals infected with West Nile Virus. J Virol 80 6982 6992

53. WalkerLMPhogatSKChan-HuiPYWagnerDPhungP 2009 Broad and potent neutralizing antibodies from an African donor reveal a new HIV-1 vaccine target. Science 326 285 289

54. ParrenPWBurtonDR 2001 The antiviral activity of antibodies in vitro and in vivo. Adv Immunol 77 195 262

55. MayJSStevensonPG 2010 Vaccination with murid herpesvirus-4 glycoprotein B reduces viral lytic replication but does not induce detectable virion neutralization. J Gen Virol 91 2542 2552

56. SinzgerCDigelMJahnG 2008 Cytomegalovirus cell tropism. Curr Top Microbiol Immunol 325 63 83

57. BenderFCSamantaMHeldweinEEde LeonMPBilmanE 2007 Antigenic and mutational analyses of herpes simplex virus glycoprotein B reveal four functional regions. J Virol 81 3827 3841

58. UtzUBrittWVuglerLMachM 1989 Identification of a neutralizing epitope on glycoprotein gp58 of human cytomegalovirus. J Virol 63 1995 2001

59. AtanasiuDWhitbeckJCde LeonMPLouHHannahBP 2010 Bimolecular complementation defines functional regions of Herpes simplex virus gB that are involved with gH/gL as a necessary step leading to cell fusion. J Virol 84 3825 3834

60. AtanasiuDSawWTCohenGHEisenbergRJ 2010 Cascade of events governing cell-cell fusion induced by herpes simplex virus glycoproteins gD, gH/gL, and gB. J Virol 84 12292 12299

61. IshibashiKTokumotoTTanabeKShirakawaHHashimotoK 2007 Association of the outcome of renal transplantation with antibody response to cytomegalovirus strain-specific glycoprotein H epitopes. Clin Infect Dis 45 60 67

62. GrundyJELuiSFSuperMBerryNJSwenyP 1988 Symptomatic cytomegalovirus infection in seropositive kidney recipients: reinfection with donor virus rather than reactivation of recipient virus. Lancet 2 132 135

63. BoppanaSBFowlerKBBrittWJStagnoSPassRF 1999 Symptomatic congenital cytomegalovirus infection in infants born to mothers with preexisting immunity to cytomegalovirus. Pediatrics 104 55 60

64. BoppanaSBRiveraLBFowlerKBMachMBrittWJ 2001 Intrauterine transmission of cytomegalovirus to infants of women with preconceptional immunity. N Engl J Med 344 1366 1371

65. AhlforsKIvarssonSAHarrisS 1999 Report on a long-term study of maternal and congenital cytomegalovirus infection in Sweden. Review of prospective studies available in the literature. Scand J Infect Dis 31 443 457

66. ScheidJFMouquetHFeldhahnNSeamanMSVelinzonK 2009 Broad diversity of neutralizing antibodies isolated from memory B cells in HIV-infected individuals. Nature 458 636 640

67. ChouSWDennisonKM 1991 Analysis of interstrain variation in cytomegalovirus glycoprotein B sequences encoding neutralization-related epitopes. J Infect Dis 163 1229 1234

68. TillerTMeffreEYurasovSTsuijiMNussenzweigMC 2008 Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. J Immunol Methods 329 112 124

69. BurkhardtCHimmeleinSBrittWWinklerTMachM 2009 Glycoprotein N subtypes of human cytomegalovirus induce a strain-specific antibody response during natural infection. J Gen Virol 90 1951 1961

70. KropffBKoedelYBrittWMachM 2010 Optimal replication of human cytomegalovirus correlates with endocytosis of glycoprotein gpUL132. J Virol 84 7039 7052

71. MachMKropffBDal-MontePBrittWJ 2000 Complex formation of human cytomegalovirus gylcoprotein M (gpUL100) and glycoprotein N (gpUl73). J Virol 74 11881 11892

72. UrbanMBrittWMachM 1992 The dominant linear neutralizing antibody-binding site of glycoprotein gp86 of human cytomegalovirus is strain specific. J Virol 66 1303 1311

73. EswarNWebbBMarti-RenomMAMadhusudhanMSEramianD 2006 Comparative protein structure modeling using Modeller. Curr Protoc Bioinformatics Chapter 5 Unit 6.1 5.6.30

74. WiedersteinMSipplMJ 2007 ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res 35 W407 W410

75. LaskowskiRAMossDSThorntonJM 1993 Main-chain bond lengths and bond angles in protein structures. J Mol Biol 231 1049 1067

76. SinzgerCHahnGDigelMKatonaRSampaioKL 2008 Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E. J Gen Virol 89 359 368