Granzyme A Produced by γδ T Cells Induces Human Macrophages to Inhibit Growth of an Intracellular Pathogen
Human γ9δ2 T cells potently inhibit pathogenic microbes, including intracellular mycobacteria, but the key inhibitory mechanism(s) involved have not been identified. We report a novel mechanism involving the inhibition of intracellular mycobacteria by soluble granzyme A. γ9δ2 T cells produced soluble factors that could pass through 0.45 µm membranes and inhibit intracellular mycobacteria in human monocytes cultured below transwell inserts. Neutralization of TNF-α in co-cultures of infected monocytes and γ9δ2 T cells prevented inhibition, suggesting that TNF-α was the critical inhibitory factor produced by γ9δ2 T cells. However, only siRNA- mediated knockdown of TNF-α in infected monocytes, but not in γ9δ2 T cells, prevented mycobacterial growth inhibition. Investigations of other soluble factors produced by γ9δ2 T cells identified a highly significant correlation between the levels of granzyme A produced and intracellular mycobacterial growth inhibition. Furthermore, purified granzyme A alone induced inhibition of intracellular mycobacteria, while knockdown of granzyme A in γ9δ2 T cell clones blocked their inhibitory effects. The inhibitory mechanism was independent of autophagy, apoptosis, nitric oxide production, type I interferons, Fas/FasL and perforin. These results demonstrate a novel microbial defense mechanism involving granzyme A-mediated triggering of TNF-α production by monocytes leading to intracellular mycobacterial growth suppression. This pathway may provide a protective mechanism relevant for the development of new vaccines and/or immunotherapies for macrophage-resident chronic microbial infections.
Vyšlo v časopise:
Granzyme A Produced by γδ T Cells Induces Human Macrophages to Inhibit Growth of an Intracellular Pathogen. PLoS Pathog 9(1): e32767. doi:10.1371/journal.ppat.1003119
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1003119
Souhrn
Human γ9δ2 T cells potently inhibit pathogenic microbes, including intracellular mycobacteria, but the key inhibitory mechanism(s) involved have not been identified. We report a novel mechanism involving the inhibition of intracellular mycobacteria by soluble granzyme A. γ9δ2 T cells produced soluble factors that could pass through 0.45 µm membranes and inhibit intracellular mycobacteria in human monocytes cultured below transwell inserts. Neutralization of TNF-α in co-cultures of infected monocytes and γ9δ2 T cells prevented inhibition, suggesting that TNF-α was the critical inhibitory factor produced by γ9δ2 T cells. However, only siRNA- mediated knockdown of TNF-α in infected monocytes, but not in γ9δ2 T cells, prevented mycobacterial growth inhibition. Investigations of other soluble factors produced by γ9δ2 T cells identified a highly significant correlation between the levels of granzyme A produced and intracellular mycobacterial growth inhibition. Furthermore, purified granzyme A alone induced inhibition of intracellular mycobacteria, while knockdown of granzyme A in γ9δ2 T cell clones blocked their inhibitory effects. The inhibitory mechanism was independent of autophagy, apoptosis, nitric oxide production, type I interferons, Fas/FasL and perforin. These results demonstrate a novel microbial defense mechanism involving granzyme A-mediated triggering of TNF-α production by monocytes leading to intracellular mycobacterial growth suppression. This pathway may provide a protective mechanism relevant for the development of new vaccines and/or immunotherapies for macrophage-resident chronic microbial infections.
Zdroje
1. BeetzS, MarischenL, KabelitzD, WeschD (2007) Human γδ T cells: candidates for the development of immunotherapeutic strategies. Immunology Research 37: 97–111.
2. MoritaCT, JinC, SarikondaG, WangH (2007) Nonpeptide antigens, presentation mechanisms, and immunological memory of human Vγ2Vδ2 T cells: discriminating friend from foe through the recognition of prenyl pyrophosphate antigens. Immunological Reviews 215: 59–76.
3. ConstantP, DavodeauF, PeyratMA, PoquetY, PuzoG, et al. (1994) Stimulation of human γδ T cells by nonpeptidic mycobacterial ligands. Science 264: 267–270.
4. AbateG, EslickJ, NewmanFK, FreySE, BelsheRB, et al. (2005) Flow-cytometric detection of vaccinia-induced memory effector CD4+, CD8+, and γδ TCR+ T cells capable of antigen-specific expansion and effector functions. Journal of Infectious Diseases 192: 1362–1371.
5. HoftDF, BabusisE, WorkuS, SpencerCT, LottenbachK, et al. (2011) Live and inactivated influenza vaccines induce similar humoral responses, but only live vaccines induce diverse T cell responses in young children. Journal of Infectious Diseases 204: 845–853.
6. HoftDF, BrownRM, RoodmanST (1998) Bacille Calmette-Guérin vaccination enhances human γδ T cell responsiveness to mycobacteria suggestive of a memory-like phenotype. Journal of Immunology 161: 1045–1054.
7. SpencerCT, AbateG, BlazevicA, HoftDF (2008) Only a subset of phosphoantigen-responsive γ9δ2 T cells mediate protective tuberculosis immunity. Journal of Immunology 181: 4471–4484.
8. DieliF, Troye-BlombergM, IvanyiJ, FournieJJ, BonnevilleM, et al. (2000) Vg9Vd2 T lymphocytes reduce the viability of intracellular Mycobacterium tuberculosis. European Journal of Immunology 30: 1512–1519.
9. DieliF, Troye-BlombergM, IvanyiJ, FournieJJ, KrenskyAM, et al. (2001) Granulysin-dependent killing of intracellular and extracellular Mycobacterium tuberculosis by Vg9Vd2 T lymphocytes. Journal of Infectious Diseases 184: 1082–1085.
10. OliaroJ, DudalS, LiautardJ, AndraultJB, LiautardJP, et al. (2005) Vg9Vd2 T cells use a combination of mechanisms to limit the spread of the pathogenic bacteria Brucella. Journal of Leukocyte Biology 77: 652–660.
11. MetkarSS, MenaaC, PardoJ, WangB, WallichR, et al. (2008) Human and mouse granzyme A induce a proinflammatory cytokine response. Immunity 29: 720–733.
12. ThurnerB, RoderC, DieckmannD, HeuerM, KruseM, et al. (1999) Generation of large numbers of fully mature and stable dendritic cells from leukapheresis products for clinical application. Journal of Immunological Methods 223: 1–15.
13. NeighbourPA, HubermanHS (1982) Sr++-induced inhibition of human natural killer (NK) cell-mediated cytotoxicity. Journal of Immunology 128: 1236–1240.
14. BlommaartEF, KrauseU, SchellensJP, Vreeling-SindelarovaH, MeijerAJ (1997) The phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 inhibit autophagy in isolated rat hepatocytes. European Journal of Biochemistry 243: 240–246.
15. SeglenPO, GordonPB (1982) 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proceedings of the National Academy of Sciences 79: 1889–1892.
16. ZhuH, FearnheadHO, CohenGM (1995) An ICE-like protease is a common mediator of apoptosis induced by diverse stimuli in human monocytic THP.1 cells. FEBS Letters 374: 303–308.
17. HannaWL, ZhangX, TurbovJ, WinklerU, HudigD, et al. (1993) Rapid purification of cationic granule proteases: application to human granzymes. Protein Expression and Purification 4: 398–404.
18. WorkuS, HoftDF (2003) Differential effects of control and antigen-specific T cells on intracellular mycobacterial growth. Infection and Immunity 71: 1763–1773.
19. PengG, GuoZ, KiniwaY, VooKs, PengW, et al. (2005) Toll-Like Receptor 8-Mediated Reversal of CD4+ Regulatory T Cell Function. Science 309: 1380–1384.
20. AllisonTJ, WinterCC, FournieJJ, BonnevilleM, GarbocziDN (2001) Structure of a human γδ-cell antigen receptor. Nature 411: 820–824.
21. BoomWH, ChervenakKA, MincekMA, EllnerJJ (1992) Role of the mononuclear phagocyte as an antigen-presenting cell for human γ/δ T cells activated by live Mycobacterium tuberculosis. Infection and Immunity 60: 3480–3488.
22. RojasRE, TorresM, FournieJJ, HardingCV, BoomWH (2002) Phosphoantigen presentation by macrophages to Mycobacterium tuberculosis-reactive Vγ9Vδ2+ T cells: modulation by chloroquine. Infection and Immunity 70: 4019–4027.
23. BurkMR, MoriL, De LiberoG (1995) Human Vγ9Vδ2 cells are stimulated in a cross-reactive fashion by a variety of phosphorylated metabolites. European Journal of Immunology 25: 2052–2058.
24. EvansPS, EndersPJ, YinC, RuckwardtTJ, MalkovskyM, et al. (2001) In vitro stimulation with a non-peptidic alkylphosphate expands cells expressing Vγ2-Jδ1.2/Vδ2 T-cell receptors. Immunology 104: 19–27.
25. MartinoA, CasettiR, SacchiA, PocciaF (2007) Central memory Vg9Vd2 T lymphocytes primed and expanded by Bacille Calmette-Guérin-infected dendritic cells kill mycobacterial-infected monocytes. Journal of Immunology 179: 3057–3064.
26. OttonesF, DornandJ, NaroeniA, LiautardJP, FaveroJ (2000) Vγ9Vδ2 T cells impair intracellular multiplication of Brucella suis in autologous monocytes through soluble factor release and contact-dependent cytotoxic effect. Journal of Immunology 165: 7133–7139.
27. WangL, KamathA, DasH, LiL, BukowskiJF (2001) Antibacterial effect of human Vg2Vd2 T cells in vivo. Journal of Clinical Investigation 108: 1349–1357.
28. LeeJ, ChoiK, OlinMR, ChoSN, MolitorTW (2004) γδ T cells in immunity induced by Mycobacterium bovis bacillus Calmette-Guérin vaccination. Infection and Immunity 72: 1504–1511.
29. TsukaguchiK, BalajiKN, BoomWH (1995) CD4+ αβ T cell and γδ T cell responses to Mycobacterium tuberculosis. Journal of Immunology 154: 1786–1796.
30. BodnarKA, SerbinaNV, FlynnJL (2001) Fate of Mycobacterium tuberculosis within murine dendritic cells. Infection and Immunity 69: 800–809.
31. CooperAM, DaltonDK, StewartTA, GriffinJP, RussellDG, et al. (1993) Disseminated tuberculosis in IFN-γ gene-disrupted mice. Journal of Experimental Medicine 178: 2243–2247.
32. FlynnJL, ChanJ, TrieboldKJ, DaltonDK, StewartTA, et al. (1993) An essential role for IFN-gamma in resistance to Mycobacterium tuberculosis infection. Journal of Experimental Medicine 178: 2249–2254.
33. OttenhoffTH, KumararatneD, CasanovaJL (1998) Novel human immunodeficiencies reveal the essential role of type-I cytokines in immunity to intracellular bacteria. Immunology Today 19: 491–494.
34. LiH, LuoK, PauzaCD (2008) TNF-alpha is a positive regulatory factor for human Vγ2 Vδ2 T cells. Journal of Immunolgy 181: 7131–7137.
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2013 Číslo 1
- Parazitičtí červi v terapii Crohnovy choroby a dalších zánětlivých autoimunitních onemocnění
- Očkování proti virové hemoragické horečce Ebola experimentální vakcínou rVSVDG-ZEBOV-GP
- Koronavirus hýbe světem: Víte jak se chránit a jak postupovat v případě podezření?
Najčítanejšie v tomto čísle
- Biosafety Level-4 Laboratories in Europe: Opportunities for Public Health, Diagnostics, and Research
- Loss and Retention of RNA Interference in Fungi and Parasites
- Innate Sensing of Chitin and Chitosan
- Make It, Take It, or Leave It: Heme Metabolism of Parasites