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A failure of TNFAIP3 negative regulation maintains sustained NF-κB activation in Sjögren’s syndrome

  • Margherita SistoEmail author
  • Sabrina Lisi
  • Dario Domenico Lofrumento
  • Giuseppe Ingravallo
  • Eugenio Maiorano
  • Massimo D’Amore
Original Paper

Abstract

Sjögren’s syndrome (SS) is characterized by the features of systemic autoimmunity and exocrine gland dysfunction and inflammation. Deregulated cytokine production is known to contribute to the etiology of SS but the underlying molecular mechanism is still remains to be unclear. TNF-α-induced protein 3 or TNFAIP3 is involved in the negative feedback regulation of nuclear factor-κB (NF-κB) signaling in response to specific pro-inflammatory stimuli in different cell types. To define the contribution of TNFAIP3 to SS, the levels of TNFAIP3 expression in human salivary gland epithelial cells (SGEC) derived from active primary SS patients were analyzed. Histological analysis was performed on paraffin-embedded human Sjögren’s samples and healthy tissues. In separate experiments, immunofluorescence staining, western blot analysis and quantitative real-time PCR for TNFAIP3 was conducted in SGEC from SS and healthy subjects. Our findings clearly demonstrate changes in levels of the protein and gene expression between healthy controls and SS patients, depicting a very weak positivity for TNFAIP3 in SS samples. TNFAIP3 was found down-regulated in SGECs derived from SS patients in comparison with controls, and the cells with down-regulated TNFAIP3 expression exhibited enhanced NF-κB activities. In addition, to investigate the role of TNFAIP3 in the activation of NF-κB, we depleted TNFAIP3 expression by siRNA in healthy SGEC after treatment with or without TNF-α. Intriguingly, the silencing of TNFAIP3 by its siRNA in healthy SGEC increased NF-κB activation that could explain the deregulated cytokines production observed in SS.

Keywords

TNFAIP3 TNF-α Sjögren’s syndrome Salivary gland NF-κB 

Notes

Acknowledgments

This work was supported by grant (No: 20216000056) from the Italian Ministry for Universities and Research. We are grateful to M.V.C. Pragnell, B.A., for critical reading of the manuscript.

References

  1. Azuma M, Motegi K, Aota K, Hayashi Y, Sato M (1997) Role of cytokines in the destruction of acinar structure in Sjögren’s syndrome salivary glands. Lab Invest 77:269–280PubMedGoogle Scholar
  2. Breckpot K, Aerts-Toegaert C, Heirman C, Peeters U, Beyaert R, Aerts JL, Thielemans K (2009) Attenuated expression of A20 markedly increases the efficacy of double-stranded RNA-activated dendritic cells as an anti-cancer vaccine. J Immunol 182:860–870PubMedGoogle Scholar
  3. Christman JW, Sadikot RT, Blackwell TS (2000) The role of nuclear factor-kappa B in pulmonary diseases. Chest 117:1482–1487PubMedCrossRefGoogle Scholar
  4. Eldor R, Yeffet A, Baum K, Doviner V, Amar D, Ben-Neriah Y, Christofori G, Peled A, Carel JC, Boitard C, Klein T, Serup P, Eizirik DL, Melloul D (2006) Conditional and specific NF-κB blockade protects pancreatic b cells from diabetogenic agents. Proc Natl Acad Sci USA 103:5072–5077PubMedCrossRefGoogle Scholar
  5. Frenzel LP, Claus R, Plume N, Schwamb J, Konermann C, Pallasch CP, Claasen J, Brinker R, Wollnik B, Plass C, Wendtner CM (2011) Sustained NF-kappa B activity in chronic lymphocytic leukemia is independent of genetic and epigenetic alterations in the TNFAIP3 (A20) locus. Int J Cancer 128:2495–24500PubMedCrossRefGoogle Scholar
  6. Fung EY, Smyth DJ, Howson JM, Cooper JD, Walker NM, Stevens H, Wicker LS, Todd JA (2009) Analysis of 17 autoimmune disease associated variants in type 1 diabetes identifi es 6q23/TNFAIP3 as a susceptibility. Genes Immun 10:188–191PubMedCrossRefGoogle Scholar
  7. Graham RR, Cotsapas C, Davies L, Hackett R, Lessard CJ, Leon JM, Burtt NP, Guiducci C, Parkin M, Gates C, Plenge RM, Behrens TW, Wither JE, Rioux JD, Fortin PR, Graham DC, Wong AK, Vyse TJ, Daly MJ, Altshuler D, Moser KL, Gaffney PM (2008) Genetic variants near TNFAIP3 on 6q23 are associated with systemic lupus erythematosus. Nat Genet 40:1059–1061PubMedCrossRefGoogle Scholar
  8. Heyninck K, Beyaert R (2005) A20 inhibits NF-kappa B activation by dual ubiquitin-editing functions. Trends Biochem Sci 30:1–4PubMedCrossRefGoogle Scholar
  9. Honma K, Tsuzuki S, Nakagawa M, Tagawa H, Nakamura S, Morishima Y, Seto M (2009) TNFAIP3/A20 functions as a novel tumor suppressor gene in several subtypes of non-Hodgkin lymphomas. Blood 114:2467–2475PubMedCrossRefGoogle Scholar
  10. Kapsogeorgou EK, Dimitriou ID, Abu-Helu RF, Moutsopoulos HM, Manoussakis MN (2001) Activation of epithelial and myoepithelial cells in the salivary glands of patients with Sjögren’s syndrome: high expression of intercellular adhesion molecule-1 (ICAM.1) in biopsy specimens and cultured cells. Clin Exp Immunol 124:126–133PubMedCrossRefGoogle Scholar
  11. Karin M (2008) The IκB kinase—a bridge between inflammation and cancer. Cell Res 18:334–342PubMedCrossRefGoogle Scholar
  12. Karin M, Ben-Neriah Y (2000) Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. Annu Rev Immunol 18:621–663PubMedCrossRefGoogle Scholar
  13. Lee EG, Boone DL, Chai S, Libby SL, Chien M, Lodolce JP, Ma A (2000) Failure to regulate TNF-induced NF-kappa B and cell death responses in A20-deficient mice. Science 289:2350–2354PubMedCrossRefGoogle Scholar
  14. Lisi S, Sisto M (2010) Effects of biological drug adalimumab on tumour necrosis factor-alpha-converting enzyme activation. Immunol Cell Biol 88:297–304PubMedCrossRefGoogle Scholar
  15. Lisi S, Sisto M, Lo frumento DD, Cucci L, Frassanito MA, Mitolo V, D’Amore M (2010) Pro-inflammatory role of Anti-Ro/SSA autoantibodies through the activation of Furin-TACE-amphiregulin axis. J Autoimmun 35:160–170PubMedCrossRefGoogle Scholar
  16. Lisi S, Sisto M, Lofrumento DD, D’Amore S, D’Amore M (2011) Advances in the understanding of the Fc gamma receptors-mediated autoantibodies uptake. Clin Exp Med 11:1–10PubMedCrossRefGoogle Scholar
  17. Margaix-Muñoz M, Bagán JV, Poveda R, Jiménez Y, Sarrión G (2009) Sjögren’s syndrome of the oral cavity Review and update. Med Oral Patol Oral Cir Bucal 14:E325–E330PubMedGoogle Scholar
  18. Musone SL, Taylor KE, Lu TT, Nititham J, Ferreira RC, Ortmann W, Shifrin N, Petri MA, Kamboh MI, Manzi S, Seldin MF, Gregersen PK, Behrens TW, Ma A, Kwok PY, Criswell LA (2008) Multiple polymorphisms in the TNFAIP3 region are independently associated with systemic lupus erythematosus. Nat Genet 40:1062–1064PubMedCrossRefGoogle Scholar
  19. Musone SL, Taylor KE, Nititham J, Chu C, Poon A, Liao W, Lam ET, Ma A, Kwok PY, Criswell LA (2011) Sequencing of TNFAIP3 and association of variants with multiple autoimmune diseases. Genes Immun (in press)Google Scholar
  20. Nair RP, Duffin KC, Helms C, Ding J, Stuart PE, Goldgar D, Gudjonsson JE, Li Y, Tejasvi T, Feng BJ, Ruether A, Schreiber S, Weichenthal M, Gladman D, Rahman P, Schrodi SJ, Prahalad S, Guthery SL, Fischer J, Liao W, Kwok PY, Menter A, Lathrop GM, Wise CA, Begovich AB, Voorhees JJ, Elder JT, Krueger GG, Bowcock AM, Abecasis GR (2009) Genome-wide scan reveals association of psoriasis with IL-23 and NF-kappa B pathways. Nat Genet 41:199–204PubMedCrossRefGoogle Scholar
  21. Nenci A, Becker C, Wullaert A, Gareus R, van Loo G, Danese S, Huth M, Nikolaev A, Neufert C, Madison B, Gumucio D, Neurath MF, Pasparakis M (2007) Epithelial NEMO links innate immunity to chronic intestinal inflammation. Nature 446:557–561PubMedCrossRefGoogle Scholar
  22. Neurath MF, Fuss I, Schurmann G, Pettersson S, Arnold K, Muller-Lobeck H, Strober W, Herfarth C, Buschenfellde KH (1998) Cytokine gene transcrition by NF-kappa B family members in patients with inflammatory bowel disease. Ann N Y Acad Sci 859:149–159PubMedCrossRefGoogle Scholar
  23. Orozco G, Hinks A, Eyre S, Ke X, Gibbons LJ, Bowes J, Flynn E, Martin P, Wellcome Trust Case Control Consortium; YEAR consortium, Wilson AG, Bax DE, Morgan AW, Emery P, Steer S, Hocking L, Reid DM, Wordsworth P, Harrison P, Thomson W, Barton A, Worthington J (2009) Combined effects of three independent SNPs greatly increase the risk estimate for RA at 6q23. Hum Mol Genet 18:2693–2699Google Scholar
  24. Plenge RM, Cotsapas C, Davies L, Price AL, de Bakker PI, Maller J, Pe’er I, Burtt NP, Blumenstiel B, DeFelice M, Parkin M, Barry R, Winslow W, Healy C, Graham RR, Neale BM, Izmailova E, Roubenoff R, Parker AN, Glass R, Karlson EW, Maher N, Hafler DA, Lee DM, Seldin MF, Remmers EF, Lee AT, Padyukov L, Alfredsson L, Coblyn J, Weinblatt ME, Gabriel SB, Purcell S, Klareskog L, Gregersen PK, Shadick NA, Daly MJ, Altshuler D (2007) Two independent alleles at 6q23 associated with risk of rheumatoid arthritis. Nat Genet 39:1477–1482PubMedCrossRefGoogle Scholar
  25. Renner F, Schmitz ML (2009) Autoregulatory feedback loops terminating the NF-κB response. Trends Biochem Sci 34:128–135PubMedCrossRefGoogle Scholar
  26. Roman-Blas JA, Jimenez SA (2006) NF kappa B as a potential therapeutic target in osteoarthritis and rheumatoid arthritis. Osteoarthr Cartil 14:839–848PubMedCrossRefGoogle Scholar
  27. Sens DA, Hintz DS, Rudisill MT, Sens MA, Spicer SS (1985) Explant culture of human submandibular gland epithelial cells: evidence for ductal origin. Lab Invest 52:559–567PubMedGoogle Scholar
  28. Sisto M, Lisi S, Lofrumento DD, Frassanito MA, Cucci L, D’Amore S, Mitolo V, D’Amore M (2009) Induction of TNF-alpha-converting enzyme-ectodomain shedding by pathogenic autoantibodies. Int Immunol 21:1341–1349PubMedCrossRefGoogle Scholar
  29. Sisto M, Lisi S, Lofrumento DD, Ingravallo G, Mitolo V, D’Amore M (2010) Expression of pro-inflammatory TACE-TNF-α-amphiregulin axis in Sjögren’s syndrome salivary glands. Histochem Cell Biol 134:345–353PubMedCrossRefGoogle Scholar
  30. Soliotis FC, Moutsopoulos HM (2004) Sjogren’s syndrome. Autoimmunity 37:305–307PubMedCrossRefGoogle Scholar
  31. Song XT, Evel-Kabler K, Shen L, Rollins L, Huang XF, Chen SY (2008) A20 is an antigen presentation attenuator, and its inhibition overcomes regulatory T cell-mediated suppression. Nat Med 14:258–265PubMedCrossRefGoogle Scholar
  32. Turer EE, Tavares RM, Mortier E, Hitotsumatsu O, Advincula R, Lee B, Shifrin N, Malynn BA, Ma A (2008) Homeostatic MyD88-dependent signals cause lethal inflammation in the absence of A20. J Exp Med 205:451–464PubMedCrossRefGoogle Scholar
  33. Valero R, Baron ML, Guerin S, Beliard S, Lelouard H, Kahn-Perles B, Vialettes B, Nguyen C, Imbert J, Naquet P (2002) A defective NF kappa B/RelB pathway in autoimmune-prone New Zealand black mice is associated with inefficient expansion of thymocyte and dendritic cells. J Immunol 169:185–192PubMedGoogle Scholar
  34. Vitali C, Bombardieri S, Jonsson R, Moutsopoulos HM, Alexander EL, Carsons SE, Daniels TE, Fox PC, Fox RI, Kassan SS, Pillemer SR, Talal N, Weisman MH (2002) Classification criteria for Sjögren’s syndrome: a revised version of the European criteria proposed by the American–European Consensus Group. European Study Group on Classification Criteria for Sjögren’s Syndrome. Ann Rheum Dis 61:554–558PubMedCrossRefGoogle Scholar
  35. Vlantis K, Pasparakis M (2010) Role of TNF in pathologies induced by nuclear factor kappaB deficiency. Curr Dir Autoimmun 11:80–93PubMedCrossRefGoogle Scholar
  36. Yamamoto Y, Gaynor RB (2001) Therapeutic potential of inhibition of the NF-kappaB pathway in the treatment of inflammation and cancer. J Clin Invest 107:135–142PubMedCrossRefGoogle Scholar
  37. Zhang SQ, Kovalenko A, Cantarella G, Wallach D (2000) Recruitment of the IKK signalosome to the p55 TNF receptor: RIP and A20 bind to NEMO (IKKgamma) upon receptor stimulation. Immunity 12:301–311PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Margherita Sisto
    • 1
    Email author
  • Sabrina Lisi
    • 1
  • Dario Domenico Lofrumento
    • 2
  • Giuseppe Ingravallo
    • 3
  • Eugenio Maiorano
    • 3
  • Massimo D’Amore
    • 4
  1. 1.Department of Human Anatomy and Histology, Laboratory of Cell BiologyUniversity of Bari Medical SchoolBariItaly
  2. 2.Department of Biological and Environmental Sciences and TechnologiesUniversity of SalentoLecceItaly
  3. 3.Department of Pathological AnatomyUniversity of Bari Medical SchoolBariItaly
  4. 4.Department of Internal Medicine and Public Medicine, Section of RheumatologyUniversity of Bari Medical SchoolBariItaly

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