Electron Tomography of HIV-1 Infection in Gut-Associated Lymphoid Tissue


Critical aspects of HIV-1 infection occur in mucosal tissues, particularly in the gut, which contains large numbers of HIV-1 target cells that are depleted early in infection. We used electron tomography (ET) to image HIV-1 in gut-associated lymphoid tissue (GALT) of HIV-1–infected humanized mice, the first three-dimensional ultrastructural examination of HIV-1 infection in vivo. Human immune cells were successfully engrafted in the mice, and following infection with HIV-1, human T cells were reduced in GALT. Virions were found by ET at all stages of egress, including budding immature virions and free mature and immature viruses. Immuno-electron microscopy verified the virions were HIV-1 and showed CD4 sequestration in the endoplasmic reticulum of infected cells. Observation of HIV-1 in infected GALT tissue revealed that most HIV-1–infected cells, identified by immunolabeling and/or the presence of budding virions, were localized to intestinal crypts with pools of free virions concentrated in spaces between cells. Fewer infected cells were found in mucosal regions and the lamina propria. The preservation quality of reconstructed tissue volumes allowed details of budding virions, including structures interpreted as host-encoded scission machinery, to be resolved. Although HIV-1 virions released from infected cultured cells have been described as exclusively mature, we found pools of both immature and mature free virions within infected tissue. The pools could be classified as containing either mostly mature or mostly immature particles, and analyses of their proximities to the cell of origin supported a model of semi-synchronous waves of virion release. In addition to HIV-1 transmission by pools of free virus, we found evidence of transmission via virological synapses. Three-dimensional EM imaging of an active infection within tissue revealed important differences between cultured cell and tissue infection models and furthered the ultrastructural understanding of HIV-1 transmission within lymphoid tissue.


Vyšlo v časopise: Electron Tomography of HIV-1 Infection in Gut-Associated Lymphoid Tissue. PLoS Pathog 10(1): e32767. doi:10.1371/journal.ppat.1003899
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003899

Souhrn

Critical aspects of HIV-1 infection occur in mucosal tissues, particularly in the gut, which contains large numbers of HIV-1 target cells that are depleted early in infection. We used electron tomography (ET) to image HIV-1 in gut-associated lymphoid tissue (GALT) of HIV-1–infected humanized mice, the first three-dimensional ultrastructural examination of HIV-1 infection in vivo. Human immune cells were successfully engrafted in the mice, and following infection with HIV-1, human T cells were reduced in GALT. Virions were found by ET at all stages of egress, including budding immature virions and free mature and immature viruses. Immuno-electron microscopy verified the virions were HIV-1 and showed CD4 sequestration in the endoplasmic reticulum of infected cells. Observation of HIV-1 in infected GALT tissue revealed that most HIV-1–infected cells, identified by immunolabeling and/or the presence of budding virions, were localized to intestinal crypts with pools of free virions concentrated in spaces between cells. Fewer infected cells were found in mucosal regions and the lamina propria. The preservation quality of reconstructed tissue volumes allowed details of budding virions, including structures interpreted as host-encoded scission machinery, to be resolved. Although HIV-1 virions released from infected cultured cells have been described as exclusively mature, we found pools of both immature and mature free virions within infected tissue. The pools could be classified as containing either mostly mature or mostly immature particles, and analyses of their proximities to the cell of origin supported a model of semi-synchronous waves of virion release. In addition to HIV-1 transmission by pools of free virus, we found evidence of transmission via virological synapses. Three-dimensional EM imaging of an active infection within tissue revealed important differences between cultured cell and tissue infection models and furthered the ultrastructural understanding of HIV-1 transmission within lymphoid tissue.


Zdroje

1. UNAIDS (2012) http://www.unaids.org/en/media/unaids/contentassets/documents/epidemiology/2012/gr2012/20121120_UNAIDS_Global_Report_2012_with_annexes_en.pdf.

2. HaaseAT (2011) Early events in sexual transmission of HIV and SIV and opportunities for interventions. Annu Rev Med 62: 127–139.

3. BrenchleyJM, DouekDC (2008) HIV infection and the gastrointestinal immune system. Mucosal Immunol 1: 23–30.

4. DouekDC, RoedererM, KoupRA (2009) Emerging concepts in the immunopathogenesis of AIDS. Annu Rev Med 60: 471–484.

5. VeazeyRS, DeMariaM, ChalifouxLV, ShvetzDE, PauleyDR, et al. (1998) Gastrointestinal tract as a major site of CD4+ T cell depletion and viral replication in SIV infection. Science 280: 427–431.

6. BrenchleyJM, SchackerTW, RuffLE, PriceDA, TaylorJH, et al. (2004) CD4+ T cell depletion during all stages of HIV disease occurs predominantly in the gastrointestinal tract. J Exp Med 200: 749–759.

7. LacknerAA, LedermanMM, RodriguezB (2012) HIV Pathogenesis: The Host. Cold Spring Harb Perspect Med 2: a007005.

8. HaaseAT (2005) Perils at mucosal front lines for HIV and SIV and their hosts. Nat Rev Immunol 5: 783–792.

9. Guy-GrandD, VassalliP (1993) Gut intraepithelial T lymphocytes. Curr Opin Immunol 5: 247–252.

10. DouekD (2007) HIV disease progression: immune activation, microbes, and a leaky gut. Top HIV Med 15: 114–117.

11. BrenchleyJM, PriceDA, SchackerTW, AsherTE, SilvestriG, et al. (2006) Microbial translocation is a cause of systemic immune activation in chronic HIV infection. Nat Med 12: 1365–1371.

12. DentonPW, GarciaJV (2012) Mucosal HIV-1 transmission and prevention strategies in BLT humanized mice. Trends in microbiology 20: 268–274.

13. BrainardDM, SeungE, FrahmN, CariappaA, BaileyCC, et al. (2009) Induction of robust cellular and humoral virus-specific adaptive immune responses in human immunodeficiency virus-infected humanized BLT mice. J Virol 83: 7305–7321.

14. DentonPW, GarciaJV (2011) Humanized mouse models of HIV infection. AIDS Rev 13: 135–148.

15. DentonPW, EstesJD, SunZ, OthienoFA, WeiBL, et al. (2008) Antiretroviral pre-exposure prophylaxis prevents vaginal transmission of HIV-1 in humanized BLT mice. PLoS Med 5: e16.

16. SunZ, DentonPW, EstesJD, OthienoFA, WeiBL, et al. (2007) Intrarectal transmission, systemic infection, and CD4+ T cell depletion in humanized mice infected with HIV-1. J Exp Med 204: 705–714.

17. LanP, TonomuraN, ShimizuA, WangS, YangYG (2006) Reconstitution of a functional human immune system in immunodeficient mice through combined human fetal thymus/liver and CD34+ cell transplantation. Blood 108: 487–492.

18. RajeshD, ZhouY, Jankowska-GanE, RoenneburgDA, DartML, et al. (2010) Th1 and Th17 immunocompetence in humanized NOD/SCID/IL2rgammanull mice. Human immunology 71: 551–559.

19. Barre-SinoussiF, ChermannJC, ReyF, NugeyreMT, ChamaretS, et al. (1983) Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science 220: 868–871.

20. GalloRC, SarinPS, GelmannEP, Robert-GuroffM, RichardsonE, et al. (1983) Isolation of human T-cell leukemia virus in acquired immune deficiency syndrome (AIDS). Science 220: 865–867.

21. OrensteinJM (2007) Replication of HIV-1 in vivo and in vitro. Ultrastruct Pathol 31: 151–167.

22. BriggsJA, RichesJD, GlassB, BartonovaV, ZanettiG, et al. (2009) Structure and assembly of immature HIV. Proc Natl Acad Sci U S A 106: 11090–11095.

23. BenjaminJ, Ganser-PornillosBK, TivolWF, SundquistWI, JensenGJ (2005) Three-dimensional structure of HIV-1 virus-like particles by electron cryotomography. J Mol Biol 346: 577–588.

24. WrightER, SchoolerJB, DingHJ, KiefferC, FillmoreC, et al. (2007) Electron cryotomography of immature HIV-1 virions reveals the structure of the CA and SP1 Gag shells. EMBO J 26: 2218–2226.

25. CarlsonLA, de MarcoA, OberwinklerH, HabermannA, BriggsJA, et al. (2010) Cryo electron tomography of native HIV-1 budding sites. PLoS Pathog 6: e1001173.

26. CarlsonLA, BriggsJA, GlassB, RichesJD, SimonMN, et al. (2008) Three-dimensional analysis of budding sites and released virus suggests a revised model for HIV-1 morphogenesis. Cell Host Microbe 4: 592–599.

27. FeltsRL, NarayanK, EstesJD, ShiD, TrubeyCM, et al. (2010) 3D visualization of HIV transfer at the virological synapse between dendritic cells and T cells. Proc Natl Acad Sci U S A 107: 13336–13341.

28. SougratR, BartesaghiA, LifsonJD, BennettAE, BessJW, et al. (2007) Electron tomography of the contact between T cells and SIV/HIV-1: implications for viral entry. PLoS Pathog 3: e63.

29. WalkerMR, PatelKK, StappenbeckTS (2009) The stem cell niche. J Pathol 217: 169–180.

30. McIntoshJR, NicastroD, MastronardeDN (2005) New views of cells in 3D: an introduction to electron tomography. Trends Cell Biol 15: 43–51.

31. GilkeyJC, StaehelinLA (1986) Advances in Ultrarapid Freezing for the Preservation of Cellular Ultrastructure. J Elect Microsc Tech 3: 177–210.

32. MorphewM, HeW, BjorkmanPJ, McIntoshJR (2008) Silver enhancement of Nanogold particles during freeze substitution for electron microscopy. J Microsc 230: 263–267.

33. LadinskyMS, HowellKE (2007) Electron tomography of immunolabeled cryosections. Methods Cell Biol 79: 543–558.

34. LutherPK, LawrenceMC, CrowtherRA (1988) A method for monitoring the collapse of plastic sections as a function of electron dose. Ultramicroscopy 24: 7–18.

35. FullerSD, WilkT, GowenBE, KrausslichHG, VogtVM (1997) Cryo-electron microscopy reveals ordered domains in the immature HIV-1 particle. Curr Biol 7: 729–738.

36. WilkT, GrossI, GowenBE, RuttenT, de HaasF, et al. (2001) Organization of immature human immunodeficiency virus type 1. J Virol 75: 759–771.

37. GanserBK, LiS, KlishkoVY, FinchJT, SundquistWI (1999) Assembly and analysis of conical models for the HIV-1 core. Science 283: 80–83.

38. HockleyDJ, WoodRD, JacobsJP, GarrettAJ (1988) Electron microscopy of human immunodeficiency virus. J Gen Virol 69: 2455–2469.

39. ZhuP, LiuJ, BessJJr, ChertovaE, LifsonJD, et al. (2006) Distribution and three-dimensional structure of AIDS virus envelope spikes. Nature 441: 847–852.

40. ChertovaE, Bess JrJWJr, CriseBJ, SowderIR, SchadenTM, et al. (2002) Envelope glycoprotein incorporation, not shedding of surface envelope glycoprotein (gp120/SU), Is the primary determinant of SU content of purified human immunodeficiency virus type 1 and simian immunodeficiency virus. J Virol 76: 5315–5325.

41. MagadanJG, Perez-VictoriaFJ, SougratR, YeY, StrebelK, et al. (2010) Multilayered mechanism of CD4 downregulation by HIV-1 Vpu involving distinct ER retention and ERAD targeting steps. PLoS Pathog 6: e1000869.

42. BriggsJA, SimonMN, GrossI, KrausslichHG, FullerSD, et al. (2004) The stoichiometry of Gag protein in HIV-1. Nat Struct Mol Biol 11: 672–675.

43. DenekaM, Pelchen-MatthewsA, BylandR, Ruiz-MateosE, MarshM (2007) In macrophages, HIV-1 assembles into an intracellular plasma membrane domain containing the tetraspanins CD81, CD9, and CD53. J Cell Biol 177: 329–341.

44. BennettAE, NarayanK, ShiD, HartnellLM, GoussetK, et al. (2009) Ion-abrasion scanning electron microscopy reveals surface-connected tubular conduits in HIV-infected macrophages. PLoS Pathog 5: e1000591.

45. Pelchen-MatthewsA, KramerB, MarshM (2003) Infectious HIV-1 assembles in late endosomes in primary macrophages. J Cell Biol 162: 443–455.

46. RaposoG, MooreM, InnesD, LeijendekkerR, Leigh-BrownA, et al. (2002) Human macrophages accumulate HIV-1 particles in MHC II compartments. Traffic 3: 718–729.

47. JollyC, KashefiK, HollinsheadM, SattentauQJ (2004) HIV-1 cell to cell transfer across an Env-induced, actin-dependent synapse. J Exp Med 199: 283–293.

48. HioeCE, ChienPCJr, LuC, SpringerTA, WangXH, et al. (2001) LFA-1 expression on target cells promotes human immunodeficiency virus type 1 infection and transmission. J Virol 75: 1077–1082.

49. RizzutoCD, SodroskiJG (1997) Contribution of virion ICAM-1 to human immunodeficiency virus infectivity and sensitivity to neutralization. J Virol 71: 4847–4851.

50. RobertsonDL, AndersonJP, BradacJA, CarrJK, FoleyB, et al. (2000) HIV-1 nomenclature proposal. Science 288: 55–56.

51. AggarwalA, IemmaTL, ShihI, NewsomeTP, McAlleryS, et al. (2012) Mobilization of HIV spread by diaphanous 2 dependent filopodia in infected dendritic cells. PLoS Pathog 8: e1002762.

52. SundquistWI, KrausslichHG (2012) HIV-1 Assembly, Budding, and Maturation. Cold Spring Harb Perspect Med 2: a006924.

53. VottelerJ, IavnilovitchE, FingrutO, ShemeshV, TaglichtD, et al. (2009) Exploring the functional interaction between POSH and ALIX and the relevance to HIV-1 release. BMC Biochem 10: 12.

54. CarlsonLA, HurleyJH (2012) In vitro reconstitution of the ordered assembly of the endosomal sorting complex required for transport at membrane-bound HIV-1 Gag clusters. Proc Natl Acad Sci U S A 109: 16928–16933.

55. BouraE, RozyckiB, ChungHS, HerrickDZ, CanagarajahB, et al. (2012) Solution structure of the ESCRT-I and -II supercomplex: implications for membrane budding and scission. Structure 20: 874–886.

56. LataS, SchoehnG, JainA, PiresR, PiehlerJ, et al. (2008) Helical structures of ESCRT-III are disassembled by VPS4. Science 321: 1354–1357.

57. HansonPI, RothR, LinY, HeuserJE (2008) Plasma membrane deformation by circular arrays of ESCRT-III protein filaments. J Cell Biol 180: 389–402.

58. von SchwedlerUK, StuchellM, MullerB, WardDM, ChungHY, et al. (2003) The protein network of HIV budding. Cell 114: 701–713.

59. BaumgartelV, IvanchenkoS, DupontA, SergeevM, WisemanPW, et al. (2011) Live-cell visualization of dynamics of HIV budding site interactions with an ESCRT component. Nat Cell Biol 13: 469–474.

60. JouvenetN, ZhadinaM, BieniaszPD, SimonSM (2011) Dynamics of ESCRT protein recruitment during retroviral assembly. Nat Cell Biol 13: 394–401.

61. YuZ, GonciarzMD, SundquistWI, HillCP, JensenGJ (2008) Cryo-EM structure of dodecameric Vps4p and its 2∶1 complex with Vta1p. J Mol Biol 377: 364–377.

62. AntonPA, MitsuyasuRT, DeeksSG, ScaddenDT, WagnerB, et al. (2003) Multiple measures of HIV burden in blood and tissue are correlated with each other but not with clinical parameters in aviremic subjects. AIDS 17: 53–63.

63. ChunTW, CarruthL, FinziD, ShenX, DiGiuseppeJA, et al. (1997) Quantification of latent tissue reservoirs and total body viral load in HIV-1 infection. Nature 387: 183–188.

64. WelschS, KepplerOT, HabermannA, AllespachI, Krijnse-LockerJ, et al. (2007) HIV-1 buds predominantly at the plasma membrane of primary human macrophages. PLoS Pathog 3: e36.

65. ChuH, WangJJ, QiM, YoonJJ, WenX, et al. (2012) The intracellular virus-containing compartments in primary human macrophages are largely inaccessible to antibodies and small molecules. PLoS One 7: e35297.

66. de MarcoA, MullerB, GlassB, RichesJD, KrausslichHG, et al. (2010) Structural analysis of HIV-1 maturation using cryo-electron tomography. PLoS Pathog 6: e1001215.

67. SwartzMA, FleuryME (2007) Interstitial flow and its effects in soft tissues. Annu Rev Biomed Eng 9: 229–256.

68. WelschS, HabermannA, JagerS, MullerB, Krijnse-LockerJ, et al. (2006) Ultrastructural analysis of ESCRT proteins suggests a role for endosome-associated tubular-vesicular membranes in ESCRT function. Traffic 7: 1551–1566.

69. BoutwellCL, RowleyCF, EssexM (2009) Reduced viral replication capacity of human immunodeficiency virus type 1 subtype C caused by cytotoxic-T-lymphocyte escape mutations in HLA-B57 epitopes of capsid protein. J Virol 83: 2460–2468.

70. MastronardeDN (2005) Automated electron microscope tomography using robust prediction of specimen movements. J Struct Biol 152: 36–51.

71. MastronardeDN (2008) Correction for non-perpendicularity of beam and tilt axis in tomographic reconstructions with the IMOD package. J Microsc 230: 212–217.

72. LadinskyMS, WuCC, McIntoshS, McIntoshJR, HowellKE (2002) Structure of the Golgi and distribution of reporter molecules at 20 degrees C reveals the complexity of the exit compartments. Mol Biol Cell 13: 2810–2825.

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2014 Číslo 1
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Eozinofilní granulomatóza s polyangiitidou
nový kurz
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa