Exposure to HIV-1 Directly Impairs Mucosal Epithelial Barrier Integrity Allowing Microbial Translocation

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While several clinical studies have shown that HIV-1 infection is associated with increased permeability of the intestinal tract, there is very little understanding of the mechanisms underlying HIV-induced impairment of mucosal barriers. Here we demonstrate that exposure to HIV-1 can directly breach the integrity of mucosal epithelial barrier, allowing translocation of virus and bacteria. Purified primary epithelial cells (EC) isolated from female genital tract and T84 intestinal cell line were grown to form polarized, confluent monolayers and exposed to HIV-1. HIV-1 X4 and R5 tropic laboratory strains and clinical isolates were seen to reduce transepithelial resistance (TER), a measure of monolayer integrity, by 30–60% following exposure for 24 hours, without affecting viability of cells. The decrease in TER correlated with disruption of tight junction proteins (claudin 1, 2, 4, occludin and ZO-1) and increased permeability. Treatment of ECs with HIV envelope protein gp120, but not HIV tat, also resulted in impairment of barrier function. Neutralization of gp120 significantly abrogated the effect of HIV. No changes to the barrier function were observed when ECs were exposed to Env defective mutant of HIV. Significant upregulation of inflammatory cytokines, including TNF-α, were seen in both intestinal and genital epithelial cells following exposure to HIV-1. Neutralization of TNF-α reversed the reduction in TERs. The disruption in barrier functions was associated with viral and bacterial translocation across the epithelial monolayers. Collectively, our data shows that mucosal epithelial cells respond directly to envelope glycoprotein of HIV-1 by upregulating inflammatory cytokines that lead to impairment of barrier functions. The increased permeability could be responsible for small but significant crossing of mucosal epithelium by virus and bacteria present in the lumen of mucosa. This mechanism could be particularly relevant to mucosal transmission of HIV-1 as well as immune activation seen in HIV-1 infected individuals.

Vyšlo v časopise: Exposure to HIV-1 Directly Impairs Mucosal Epithelial Barrier Integrity Allowing Microbial Translocation. PLoS Pathog 6(4): e32767. doi:10.1371/journal.ppat.1000852
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000852

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Zdroje

1. ShattockRJ

HaynesBF

PulendranB

FloresJ

EsparzaJ

2008 Improving defences at the portal of HIV entry: mucosal and innate immunity. PLoS Med 5 e81 doi:10.1371/journal.pmed.0050081

2. WiraCR

Grant-TschudyKS

Crane-GodreauMA

2005 Epithelial cells in the female reproductive tract: a central role as sentinels of immune protection. Am J Reprod Immunol 53 65 76

3. VrolingAB

FokkensWJ

van DrunenCM

2008 How epithelial cells detect danger: aiding the immune response. Allergy 63 1110 1123

4. YuQH

YangQ

2009 Diversity of tight junctions (TJs) between gastrointestinal epithelial cells and their function in maintaining the mucosal barrier. Cell Biol Int 33 78 82

5. ShacklettBL

CritchfieldJW

FerreAL

HayesTL

2009 Mucosal T-cell responses to HIV: responding at the front lines. J Intern Med 265 58 66

6. MoserLA

CarterM

Schultz-CherryS

2007 Astrovirus increases epithelial barrier permeability independently of viral replication. J Virol 81 11937 11945

7. ChenML

GeZ

FoxJG

SchauerDB

2006 Disruption of tight junctions and induction of proinflammatory cytokine responses in colonic epithelial cells by Campylobacter jejuni. Infect Immun 74 6581 6589

8. JuY

WangT

LiY

XinW

WangS

2007 Coxsackievirus B3 affects endothelial tight junctions: possible relationship to ZO-1 and F-actin, as well as p38 MAPK activity. Cell Biol Int 31 1207 1213

9. NavaP

LopezS

AriasCF

IslasS

Gonzalez-MariscalL

2004 The rotavirus surface protein VP8 modulates the gate and fence function of tight junctions in epithelial cells. J Cell Sci 117 5509 5519

10. SandersDS

2005 Mucosal integrity and barrier function in the pathogenesis of early lesions in Crohn's disease. J Clin Pathol 58 568 572

11. BrenchleyJM

DouekDC

2008 HIV infection and the gastrointestinal immune system. Mucosal Immunol 1 23 30

12. HladikF

HopeTJ

2009 HIV infection of the genital mucosa in women. Curr HIV/AIDS Rep 6 20 28

13. HladikF

McElrathMJ

2008 Setting the stage: host invasion by HIV. Nat Rev Immunol 8 447 457

14. EppleHJ

SchneiderT

TroegerH

KunkelD

AllersK

2009 Impairment of the intestinal barrier is evident in untreated but absent in suppressively treated HIV-infected patients. Gut 58 220 227

15. LimSG

MenziesIS

LeeCA

JohnsonMA

PounderRE

1993 Intestinal permeability and function in patients infected with human immunodeficiency virus. A comparison with coeliac disease. Scand J Gastroenterol 28 573 580

16. KeatingJ

BjarnasonI

SomasundaramS

MacphersonA

FrancisN

1995 Intestinal absorptive capacity, intestinal permeability and jejunal histology in HIV and their relation to diarrhoea. Gut 37 623 629

17. SharpstoneD

NeildP

CraneR

TaylorC

HodgsonC

1999 Small intestinal transit, absorption, and permeability in patients with AIDS with and without diarrhoea. Gut 45 70 76

18. StockmannM

SchmitzH

FrommM

SchmidtW

PauliG

2000 Mechanisms of epithelial barrier impairment in HIV infection. Ann N Y Acad Sci 915 293 303

19. StockmannM

FrommM

SchmitzH

SchmidtW

RieckenEO

1998 Duodenal biopsies of HIV-infected patients with diarrhoea exhibit epithelial barrier defects but no active secretion. Aids 12 43 51

20. BrenchleyJM

DouekDC

2008 The mucosal barrier and immune activation in HIV pathogenesis. Curr Opin HIV AIDS 3 356 361

21. ReddAD

DabitaoD

BreamJH

CharvatB

LaeyendeckerO

2009 Microbial translocation, the innate cytokine response, and HIV-1 disease progression in Africa. Proc Natl Acad Sci U S A 106 6718 6723

22. BrenchleyJ

PriceD

DouekD

2006 HIV Disease: Fallout from a mucosal catastrophe? Nat Immunol 7 235 239

23. HazenbergMD

OttoSA

van BenthemBH

RoosMT

CoutinhoRA

2003 Persistent immune activation in HIV-1 infection is associated with progression to AIDS. Aids 17 1881 1888

24. GiorgiJV

HultinLE

McKeatingJA

JohnsonTD

OwensB

1999 Shorter survival in advanced human immunodeficiency virus type 1 infection is more closely associated with T lymphocyte activation than with plasma virus burden or virus chemokine coreceptor usage. J Infect Dis 179 859 870

25. BrenchleyJM

PriceDA

SchackerTW

AsherTE

SilvestriG

2006 Microbial translocation is a cause of systemic immune activation in chronic HIV infection. Nat Med 12 1365 1371

26. VelardeG

Ait-AissaS

GilletC

RogerieuxF

LambreC

1999 Use of Transepithelial Electrical resistance in the study of Pentachlorophenol Toxicity. Toxicology in vitro 13 723 727

27. AnnunziataP

2003 Blood-brain barrier changes during invasion of the central nervous system by HIV-1. Old and new insights into the mechanism. J Neurol 250 901 906

28. KanmogneGD

SchallK

LeibhartJ

KnipeB

GendelmanHE

2007 HIV-1 gp120 compromises blood-brain barrier integrity and enhances monocyte migration across blood-brain barrier: implication for viral neuropathogenesis. J Cereb Blood Flow Metab 27 123 134

29. PuH

TianJ

AndrasIE

HayashiK

FloraG

2005 HIV-1 Tat protein-induced alterations of ZO-1 expression are mediated by redox-regulated ERK 1/2 activation. J Cereb Blood Flow Metab 25 1325 1335

30. OttDE

ChertovaEN

BuschLK

CorenLV

GagliardiTD

1999 Mutational analysis of the hydrophobic tail of the human immunodeficiency virus type 1 p6(Gag) protein produces a mutant that fails to package its envelope protein. J Virol 73 19 28

31. FichorovaRN

DesaiPJ

GibsonFCI

GencoCA

2001 Distinct proinflammatory host responses to Neisseria gonorrhoeae infection in immortalized human cervical and vaginal epithelial cells. Infec Imm 69 5840 5848

32. Al-SadiR

BoivinM

MaT

2009 Mechanism of cytokine modulation of epithelial tight junction barrier. Front Biosci 14 2765 2778

33. YeD

MaI

MaTY

2006 Molecular mechanism of tumor necrosis factor-alpha modulation of intestinal epithelial tight junction barrier. Am J Physiol Gastrointest Liver Physiol 290 G496 504

34. SchmitzH

RokosK

FlorianP

GitterAH

FrommM

2002 Supernatants of HIV-infected immune cells affect barrier function of human HT-29/B6 intestinal epithelial cells. AIDS 16 983 991

35. SteffenM

ReineckerHC

PetersenJ

DoehnC

PflugerI

1993 Differences in cytokine secretion by intestinal mononuclear cells, peripheral blood monocytes and alveolar macrophages from HIV-infected patients. Clin Exp Immunol 91 30 36

36. SnijdersF

van DeventerSJ

BartelsmanJF

den OtterP

JansenJ

1995 Diarrhoea in HIV-infected patients: no evidence of cytokine-mediated inflammation in jejunal mucosa. Aids 9 367 373

37. BrabersNA

NottetHS

2006 Role of the pro-inflammatory cytokines TNF-alpha and IL-1beta in HIV-associated dementia. Eur J Clin Invest 36 447 458

38. BaiL

ZhangZ

ZhangH

LiX

YuQ

2008 HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study. BMC Infect Dis 8 77

39. KagnoffMF

EckmannL

1997 Epithelial cells as sensors for microbial infection. J Clin Invest 100 6 10

40. MaTY

IwamotoGK

HoaNT

AkotiaV

PedramA

2004 TNF-alpha-induced increase in intestinal epithelial tight junction permeability requires NF-kappa B activation. Am J Physiol Gastrointest Liver Physiol 286 G367 376

41. SharkeyDJ

MacphersonAM

TremellenKP

RobertsonSA

2007 Seminal plasma differentially regulates inflammatory cytokine gene expression in human cervical and vaginal epithelial cells. Mol Hum Reprod 13 491 501

42. GutscheS

von WolffM

StrowitzkiT

ThalerCJ

2003 Seminal plasma induces mRNA expression of IL-1beta, IL-6 and LIF in endometrial epithelial cells in vitro. Mol Hum Reprod 9 785 791

43. MunchJ

RuckerE

StandkerL

AdermannK

GoffinetC

2007 Semen-derived amyloid fibrils drastically enhance HIV infection. Cell 131 1059 1071

44. PlanchonS

FiocchiC

TakafujiV

RocheJK

1999 Transforming growth factor-beta1 preserves epithelial barrier function: identification of receptors, biochemical intermediates, and cytokine antagonists. J Cell Physiol 181 55 66

45. PilcherCD

JoakiG

HoffmanIF

MartinsonFE

MapanjeC

2007 Amplified transmission of HIV-1: comparison of HIV-1 concentrations in semen and blood during acute and chronic infection. Aids 21 1723 1730

46. TachetA

DulioustE

SalmonD

De AlmeidaM

RivallandS

1999 Detection and quantification of HIV-1 in semen: identification of a subpopulation of men at high potential risk of viral sexual transmission. Aids 13 823 831

47. ShethPM

KovacsC

KemalKS

JonesRB

RaboudJM

2009 Persistent HIV RNA shedding in semen despite effective antiretroviral therapy. Aids 23 2050 2054

48. KaulR

PettengellC

ShethPM

SunderjiS

BiringerA

2008 The genital tract immune milieu: an important determinant of HIV susceptibility and secondary transmission. J Reprod Immunol 77 32 40

49. MacDonaldEM

SavoyA

GillgrassA

FernandezS

SmiejaM

2007 Susceptibility of Human Female Primary Genital Epithelial Cells to Herpes Simplex Virus, Type-2 and the Effect of TLR3 Ligand and Sex Hormones on Infection. Biol Reprod 77 1049 1059

50. McKayDM

SinghPK

1997 Superantigen activation of immune cells evokes epithelial (T84) transport and barrier abnormalities via IFN-gamma and TNF alpha: inhibition of increased permeability, but not diminished secretory responses by TGF-beta2. J Immunol 159 2382 2390

51. KimptonJ

EmermanM

1992 Detection of replication-competent and pseudotyped human immunodeficiency virus with a sensitive cell line on the basis of activation of an integrated beta-galactosidase gene. J Virol 66 2232 2239

52. SchmittgenTD

LivakKJ

2008 Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3 1101 1108

53. NazliA

YaoXD

SmiejaM

RosenthalKL

AshkarAA

2009 Differential induction of innate anti-viral responses by TLR ligands against Herpes simplex virus, type 2, infection in primary genital epithelium of women. Antiviral Res 81 103 112