Molecular Evolution of Broadly Neutralizing Llama Antibodies to the CD4-Binding Site of HIV-1

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Developing a vaccine against HIV-1 is a priority, but it remains unclear whether immunizations in humans can elicit potent broadly neutralizing antibodies able to prevent HIV-1 transmission. Llamas possess heavy chain only antibodies and conventional heavy and light chain antibodies. We previously reported the heavy chain only antibody J3, which potently neutralizes more than 95% of HIV strains, and was induced by immunization. Here we immunized two further llamas and elicited three novel broadly neutralizing heavy chain only antibodies, which were identified by high-throughput screening. These neutralizing llama antibodies target different areas of the CD4-binding site of the virus, therefore breadth and potency are increased when they are used in combination. To gain greater understanding of how the llama immunizations worked, deep sequencing of the HIV binding region of the antibodies was performed. This revealed that the antibodies were matured fully only in response to the protein immunogens. Furthermore, the VHH elicited in different animals, while sharing functional hallmarks, were encoded by distinct sequences and thus could not have been identified by a deep sequencing analysis alone. Our results show that immunization can potentially induce protective antibodies in llamas and provide a method to more extensively evaluate immunization studies.

Vyšlo v časopise: Molecular Evolution of Broadly Neutralizing Llama Antibodies to the CD4-Binding Site of HIV-1. PLoS Pathog 10(12): e32767. doi:10.1371/journal.ppat.1004552
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004552

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Zdroje

1. Abdool KarimQ, Abdool KarimSS, FrohlichJA, GroblerAC, BaxterC, et al. (2010) Effectiveness and safety of tenofovir gel, an antiretroviral microbicide, for the prevention of HIV infection in women. Science 329 : 1168–1174.

2. LagenaurLA, Sanders-BeerBE, BrichacekB, PalR, LiuX, et al. (2011) Prevention of vaginal SHIV transmission in macaques by a live recombinant Lactobacillus. Mucosal Immunol 4 : 648–657.

3. BalazsAB, ChenJ, HongCM, RaoDS, YangL, et al. (2011) Antibody-based protection against HIV infection by vectored immunoprophylaxis. Nature 481 : 81–84.

4. BalazsAB, OuyangY, HongCM, ChenJ, NguyenSM, et al. (2014) Vectored immunoprophylaxis protects humanized mice from mucosal HIV transmission. Nat Med 20 : 296–300.

5. HaynesBF, GilbertPB, McElrathMJ, Zolla-PaznerS, TomarasGD, et al. (2012) Immune-correlates analysis of an HIV-1 vaccine efficacy trial. N Engl J Med 366 : 1275–1286.

6. MontefioriDC, KarnasutaC, HuangY, AhmedH, GilbertP, et al. (2012) Magnitude and Breadth of the Neutralizing Antibody Response in the RV144 and Vax003 HIV-1 Vaccine Efficacy Trials. J Infect Dis 206 : 431–41.

7. Rerks-NgarmS, PitisuttithumP, NitayaphanS, KaewkungwalJ, ChiuJ, et al. (2009) Vaccination with ALVAC and AIDSVAX to prevent HIV-1 infection in Thailand. N Engl J Med 361 : 2209–2220.

8. HessellAJ, RakaszEG, PoignardP, HangartnerL, LanducciG, et al. (2009) Broadly neutralizing human anti-HIV antibody 2G12 is effective in protection against mucosal SHIV challenge even at low serum neutralizing titers. PLoS Pathog 5: e1000433.

9. MascolaJR, LewisMG, StieglerG, HarrisD, VanCottTC, et al. (1999) Protection of Macaques against pathogenic simian/human immunodeficiency virus 89.6PD by passive transfer of neutralizing antibodies. J Virol 73 : 4009–4018.

10. MascolaJR, StieglerG, VanCottTC, KatingerH, CarpenterCB, et al. (2000) Protection of macaques against vaginal transmission of a pathogenic HIV-1/SIV chimeric virus by passive infusion of neutralizing antibodies. Nat Med 6 : 207–210.

11. MoldtB, RakaszEG, SchultzN, Chan-HuiPY, SwiderekK, et al. (2012) Highly potent HIV-specific antibody neutralization in vitro translates into effective protection against mucosal SHIV challenge in vivo. Proc Natl Acad Sci U S A 109 : 18921–18925.

12. WatkinsJD, SiddappaNB, LakhasheSK, HumbertM, SholukhA, et al. (2011) An anti-HIV-1 V3 loop antibody fully protects cross-clade and elicits T-cell immunity in macaques mucosally challenged with an R5 clade C SHIV. PLoS One 6: e18207.

13. WatkinsJD, SholukhAM, MukhtarMM, SiddappaNB, LakhasheSK, et al. (2013) Anti-HIV IgA isotypes: differential virion capture and inhibition of transcytosis are linked to prevention of mucosal R5 SHIV transmission. AIDS 27: F13–20.

14. BarouchDH, WhitneyJB, MoldtB, KleinF, OliveiraTY, et al. (2013) Therapeutic efficacy of potent neutralizing HIV-1-specific monoclonal antibodies in SHIV-infected rhesus monkeys. Nature 503 : 224–228.

15. PlotkinSA (2008) Vaccines: correlates of vaccine-induced immunity. Clin Infect Dis 47 : 401–409.

16. MoorePL, GrayES, WibmerCK, BhimanJN, NonyaneM, et al. (2012) Evolution of an HIV glycan-dependent broadly neutralizing antibody epitope through immune escape. Nat Med 18 : 1688–1692.

17. WibmerCK, BhimanJN, GrayES, TumbaN, Abdool KarimSS, et al. (2013) Viral escape from HIV-1 neutralizing antibodies drives increased plasma neutralization breadth through sequential recognition of multiple epitopes and immunotypes. PLoS Pathog 9: e1003738.

18. PoignardP, SabbeR, PicchioGR, WangM, GuliziaRJ, et al. (1999) Neutralizing antibodies have limited effects on the control of established HIV-1 infection in vivo. Immunity 10 : 431–438.

19. Doria-RoseNA, LouderMK, YangZ, O'DellS, NasonM, et al. (2012) HIV-1 Neutralization Coverage Is Improved by Combining Monoclonal Antibodies That Target Independent Epitopes. J Virol 86 : 3393–7.

20. ScheidJF, MouquetH, UeberheideB, DiskinR, KleinF, et al. (2011) Sequence and structural convergence of broad and potent HIV antibodies that mimic CD4 binding. Science 333 : 1633–1637.

21. LiaoHX, BonsignoriM, AlamSM, McLellanJS, TomarasGD, et al. (2013) Vaccine induction of antibodies against a structurally heterogeneous site of immune pressure within HIV-1 envelope protein variable regions 1 and 2. Immunity 38 : 176–186.

22. ForsellMN, SoldemoM, DosenovicP, WyattRT, KarlssonMC, et al. (2013) Independent expansion of epitope-specific plasma cell responses upon HIV-1 envelope glycoprotein immunization. J Immunol 191 : 44–51.

23. TranK, PoulsenC, GuenagaJ, de ValN, WilsonR, et al. (2014) Vaccine-elicited primate antibodies use a distinct approach to the HIV-1 primary receptor binding site informing vaccine redesign. Proc Natl Acad Sci U S A 111: E738–747.

24. CarbonettiS, OliverBG, GlennJ, StamatatosL, SatherDN (2014) Soluble HIV-1 envelope immunogens derived from an elite neutralizer elicit cross-reactive V1V2 antibodies and low potency neutralizing antibodies. PLoS One 9: e86905.

25. McCoyLE, WeissRA (2013) Neutralizing antibodies to HIV-1 induced by immunization. J Exp Med 210 : 209–223.

26. McCoyLE, QuigleyAF, StrokappeNM, Bulmer-ThomasB, SeamanMS, et al. (2012) Potent and broad neutralization of HIV-1 by a llama antibody elicited by immunization. J Exp Med 209 : 1091–1103.

27. Hamers-CastermanC, AtarhouchT, MuyldermansS, RobinsonG, HamersC, et al. (1993) Naturally occurring antibodies devoid of light chains. Nature 363 : 446–448.

28. GorlaniA, BrouwersJ, McConvilleC, van der BijlP, MalcolmK, et al. (2011) Llama antibody fragments have good potential for application as HIV type 1 topical microbicides. AIDS Res Hum Retroviruses 28 : 198–205.

29. PantN, MarcotteH, HermansP, BezemerS, FrenkenL, et al. (2011) Lactobacilli producing bispecific llama-derived anti-rotavirus proteins in vivo for rotavirus-induced diarrhea. Future Microbiol 6 : 583–593.

30. SeamanMS, JanesH, HawkinsN, GrandpreLE, DevoyC, et al. (2010) Tiered categorization of a diverse panel of HIV-1 Env pseudoviruses for assessment of neutralizing antibodies. J Virol 84 : 1439–1452.

31. StrokappeN, SzynolA, Aasa-ChapmanM, GorlaniA, Forsman QuigleyA, et al. (2012) Llama antibody fragments recognizing various epitopes of the CD4bs neutralize a broad range of HIV-1 subtypes A, B and C. PLoS One 7: e33298.

32. Lutje HulsikD, LiuYY, StrokappeNM, BattellaS, El KhattabiM, et al. (2013) A gp41 MPER-specific llama VHH requires a hydrophobic CDR3 for neutralization but not for antigen recognition. PLoS Pathog 9: e1003202.

33. WuX, YangZY, LiY, HogerkorpCM, SchiefWR, et al (2010) Rational design of envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1. Science 329 : 856–861.

34. ZhouT, GeorgievI, WuX, YangZY, DaiK, et al. (2010) Structural basis for broad and potent neutralization of HIV-1 by antibody VRC01. Science 329 : 811–817.

35. KwongPD, DoyleML, CasperDJ, CicalaC, LeavittSA, et al. (2002) HIV-1 evades antibody-mediated neutralization through conformational masking of receptor-binding sites. Nature 420 : 678–682.

36. ForsmanA, BeirnaertE, Aasa-ChapmanMM, HoorelbekeB, HijaziK, et al. (2008) Llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (HIV-1)-neutralizing properties and high affinity for HIV-1 gp120. J Virol 82 : 12069–12081.

37. MatzJ, KesslerP, BouchetJ, CombesO, RamosOH, et al. (2012) Straightforward Selection of Broadly Neutralizing Single-Domain Antibodies Targeting the Conserved CD4 and Coreceptor Binding Sites of HIV-1 gp120. J Virol 87 : 1137–1149.

38. DiskinR, ScheidJF, MarcovecchioPM, WestAPJr, KleinF, et al. (2011) Increasing the Potency and Breadth of an HIV Antibody by Using Structure-Based Rational Design. Science 334 : 1289–93.

39. CortiD, LangedijkJP, HinzA, SeamanMS, VanzettaF, et al. (2010) Analysis of memory B cell responses and isolation of novel monoclonal antibodies with neutralizing breadth from HIV-1-infected individuals. PLoS One 5: e8805.

40. FalkowskaE, RamosA, FengY, ZhouT, MoquinS, et al. (2012) PGV04, an HIV-1 gp120 CD4 binding site antibody, is broad and potent in neutralization but does not induce conformational changes characteristic of CD4. J Virol 86 : 4394–4403.

41. WestAPJr, DiskinR, NussenzweigMC, BjorkmanPJ (2012) Structural basis for germ-line gene usage of a potent class of antibodies targeting the CD4-binding site of HIV-1 gp120. Proc Natl Acad Sci U S A 109: E2083–2090.

42. WalkerLM, PhogatSK, Chan-HuiPY, WagnerD, PhungP, et al. (2009) Broad and potent neutralizing antibodies from an African donor reveal a new HIV-1 vaccine target. Science 326 : 285–289.

43. WalkerLM, HuberM, DooresKJ, FalkowskaE, PejchalR, et al. (2011) Broad neutralization coverage of HIV by multiple highly potent antibodies. Nature 477 : 466–470.

44. Doria-RoseNA, SchrammCA, GormanJ, MoorePL, BhimanJN, et al. (2014) Developmental pathway for potent V1V2-directed HIV-neutralizing antibodies. Nature 509 : 55–62.

45. McCoyLE, GroppelliE, BlanchetotC, De HaardH, VerripsC, et al. (2014) Neutralisation of HIV-1 cell-cell spread by human and llama antibodies. Retroviroloy In Press.

46. XiaoX, ChenW, FengY, ZhuZ, PrabakaranP, et al. (2009) Germline-like predecessors of broadly neutralizing antibodies lack measurable binding to HIV-1 envelope glycoproteins: implications for evasion of immune responses and design of vaccine immunogens. Biochem Biophys Res Commun 390 : 404–409.

47. McGuireAT, GlennJA, LippyA, StamatatosL (2013) Diverse recombinant HIV-1 Envs fail to activate B cells expressing the germline B cell receptors of the broadly neutralizing anti-HIV-1 antibodies PG9 and 447-52D. J Virol 88 : 2645–2657.

48. HootS, McGuireAT, CohenKW, StrongRK, HangartnerL, et al. (2013) Recombinant HIV envelope proteins fail to engage germline versions of anti-CD4bs bNAbs. PLoS Pathog 9: e1003106.

49. JardineJ, JulienJP, MenisS, OtaT, KalyuzhniyO, et al. (2013) Rational HIV immunogen design to target specific germline B cell receptors. Science 340 : 711–716.

50. ScharfL, WestAPJr, GaoH, LeeT, ScheidJF, et al. (2013) Structural basis for HIV-1 gp120 recognition by a germ-line version of a broadly neutralizing antibody. Proc Natl Acad Sci U S A 110 : 6049–6054.

51. McGuireAT, HootS, DreyerAM, LippyA, StuartA, et al. (2013) Engineering HIV envelope protein to activate germline B cell receptors of broadly neutralizing anti-CD4 binding site antibodies. J Exp Med 210 : 655–663.

52. LiaoHX, LynchR, ZhouT, GaoF, AlamSM, et al. (2013) Co-evolution of a broadly neutralizing HIV-1 antibody and founder virus. Nature 496 : 469–476.

53. ZhouT, ZhuJ, WuX, MoquinS, ZhangB, et al. (2014) Multidonor analysis reveals structural elements, genetic determinants, and maturation pathway for HIV-1 neutralization by VRC01-class antibodies. Immunity 39 : 245–258.

54. BonsignoriM, MontefioriDC, WuX, ChenX, HwangKK, et al. (2012) Two distinct broadly neutralizing antibody specificities of different clonal lineages in a single HIV-1-infected donor: implications for vaccine design. J Virol 86 : 4688–4692.

55. SundlingC, ZhangZ, PhadGE, ShengZ, WangY, et al. (2014) Single-Cell and Deep Sequencing of IgG-Switched Macaque B Cells Reveal a Diverse Ig Repertoire following Immunization. J Immunol 192 : 3637–3644.

56. KumaranJ, MackenzieCR, Arbabi-GhahroudiM (2012) Semiautomated panning of naive camelidae libraries and selection of single-domain antibodies against peptide antigens. Methods Mol Biol 911 : 105–124.

57. Bashford-RogersRJ, PalserAL, HuntlyBJ, RanceR, VassiliouGS, et al. (2013) Network properties derived from deep sequencing of human B-cell receptor repertoires delineate B-cell populations. Genome Res 23 : 1874–84.

58. KleinF, Halper-StrombergA, HorwitzJA, GruellH, ScheidJF, et al. (2012) HIV therapy by a combination of broadly neutralizing antibodies in humanized mice. Nature 492 : 118–122.

59. GrafM, BojakA, DemlL, BielerK, WolfH, et al. (2000) Concerted action of multiple cis-acting sequences is required for Rev dependence of late human immunodeficiency virus type 1 gene expression. J Virol 74 : 10822–10826.

60. Csardi G, Nepusz T (2006) The igraph software package for complex network research, InterJournal, Complex Systems 1695. http://igraphsfnet.

61. AltschulSF, GishW, MillerW, MyersEW, LipmanDJ (1990) Basic local alignment search tool. J Mol Biol 215 : 403–410.

62. Wickham H (2009) ggplot2: elegant graphics for data analysis. New York: Springer.