Regulation of NF-κB- and STAT1-mediated plasmacytoid dendritic cell functions by A20

Autoři: Pham Ngoc Duy aff001;  Nguyen Thu Thuy aff002;  Bui Kieu Trang aff003;  Nguyen Hoang Giang aff003;  Nguyen Thi Hong Van aff004;  Nguyen Thi Xuan aff003
Působiště autorů: Faculty of Biotechnology, Vietnam National University of Agriculture, Trau Quy, Gia Lam, Hanoi, Vietnam aff001;  Institute of Biomedicine and Pharmacy, Vietnam Military Medical University, Ha Dong, Hanoi, Vietnam aff002;  Institute of Genome Research, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam aff003;  Department of Genetics, Faculty of Biology, VNU University of Science, Thanh Xuan, Hanoi, Vietnam aff004;  Graduate University of Science and Technology, Vietnam Academy of Science and Technology, Cau Giay, Ha Noi, Vietnam aff005
Vyšlo v časopise: PLoS ONE 14(9)
Kategorie: Research Article
doi: 10.1371/journal.pone.0222697


Dendritic cells (DCs) are professional antigen presenting cells involved in the induction of T cell-mediated adaptive immunity. Plasmacytoid DCs (pDCs) originate from lymphoid precursors and produce type I interferons (IFNs) in response to pathogens. A20 is considered as a negative regulator of toll-like receptor (TLR) signaling pathways, in which Toxoplasma gondii- derived profilin (TgPRF) is a TLR11/12 ligand recognised by DCs to stimulate their maturation/activation. Little is known about contributions of A20 to changes in biological properties of pDCs. The present study, therefore, explored whether pDC functions are influenced by A20. To this end, bone marrow cells were isolated and cultured with Flt3L to attain CD8DCs, CD11bDCs and pDCs and followed by challenge with TgPRP in the presence or absence of A20 siRNA. Expression of maturation markers were analysed by flow cytometry, and secretion of inflammatory cytokines by ELISA, cell migration by a transwell migration assay and expression of signalling molecules by western blotting. As a result, treatment with A20 siRNA enhanced activations of IκB-α and STAT-1, leading to increases in expressions of maturation markers and cytokine productions as well as migration of TgPRP-treated pDCs, while mature CD11bDCs produced at higher levels of TNF-α and IL-6 only. In addition, functions of CD8DCs remained unaltered following A20 silencing. The effects of A20 on pDC maturation and activation were completely abolished by IKK inhibitor and partially blunted by fludarabine. In conclusion, the inhibitory effects of A20 on pDC functions are expected to affect the immune response in T. gondii infection.

Klíčová slova:

Cell migration – Cytokines – Secretion – Signal inhibition – Small interfering RNAs – Toxoplasma gondii – Toll-like receptors – Arithmetic


1. Kohli K, Janssen A, Forster R. Plasmacytoid dendritic cells induce tolerance predominantly by cargoing antigen to lymph nodes. Eur J Immunol. 2016;46(11):2659–68. doi: 10.1002/eji.201646359 27592607.

2. de Heer HJ, Hammad H, Soullie T, Hijdra D, Vos N, Willart MA, et al. Essential role of lung plasmacytoid dendritic cells in preventing asthmatic reactions to harmless inhaled antigen. J Exp Med. 2004;200(1):89–98. doi: 10.1084/jem.20040035 15238608.

3. Wang Y, McGivern DR, Cheng L, Li G, Lemon SM, Niu J, et al. Ribavirin Contributes to Hepatitis C Virus Suppression by Augmenting pDC Activation and Type 1 IFN Production. PLoS One. 2015;10(8):e0135232. doi: 10.1371/journal.pone.0135232 26274905.

4. Sozzani S, Del Prete A, Bosisio D. Dendritic cell recruitment and activation in autoimmunity. J Autoimmun. 2017;85:126–40. doi: 10.1016/j.jaut.2017.07.012 28774715.

5. Carrington EM, Zhang JG, Sutherland RM, Vikstrom IB, Brady JL, Soo P, et al. Prosurvival Bcl-2 family members reveal a distinct apoptotic identity between conventional and plasmacytoid dendritic cells. Proc Natl Acad Sci U S A. 2015;112(13):4044–9. doi: 10.1073/pnas.1417620112 25775525.

6. Roach JC, Glusman G, Rowen L, Kaur A, Purcell MK, Smith KD, et al. The evolution of vertebrate Toll-like receptors. Proc Natl Acad Sci U S A. 2005;102(27):9577–82. doi: 10.1073/pnas.0502272102 15976025.

7. Salazar Gonzalez RM, Shehata H, O'Connell MJ, Yang Y, Moreno-Fernandez ME, Chougnet CA, et al. Toxoplasma gondii- derived profilin triggers human toll-like receptor 5-dependent cytokine production. J Innate Immun. 2014;6(5):685–94. doi: 10.1159/000362367 24861338.

8. Koblansky AA, Jankovic D, Oh H, Hieny S, Sungnak W, Mathur R, et al. Recognition of profilin by Toll-like receptor 12 is critical for host resistance to Toxoplasma gondii. Immunity. 2013;38(1):119–30. doi: 10.1016/j.immuni.2012.09.016 23246311.

9. Aliberti J, Valenzuela JG, Carruthers VB, Hieny S, Andersen J, Charest H, et al. Molecular mimicry of a CCR5 binding-domain in the microbial activation of dendritic cells. Nat Immunol. 2003;4(5):485–90. doi: 10.1038/ni915 12665855.

10. Pepper M, Dzierszinski F, Wilson E, Tait E, Fang Q, Yarovinsky F, et al. Plasmacytoid dendritic cells are activated by Toxoplasma gondii to present antigen and produce cytokines. J Immunol. 2008;180(9):6229–36. doi: 10.4049/jimmunol.180.9.6229 18424745.

11. Liu CH, Fan YT, Dias A, Esper L, Corn RA, Bafica A, et al. Cutting edge: dendritic cells are essential for in vivo IL-12 production and development of resistance against Toxoplasma gondii infection in mice. J Immunol. 2006;177(1):31–5. doi: 10.4049/jimmunol.177.1.31 16785494.

12. Yarovinsky F, Zhang D, Andersen JF, Bannenberg GL, Serhan CN, Hayden MS, et al. TLR11 activation of dendritic cells by a protozoan profilin-like protein. Science. 2005;308(5728):1626–9. doi: 10.1126/science.1109893 15860593.

13. Scanga CA, Aliberti J, Jankovic D, Tilloy F, Bennouna S, Denkers EY, et al. Cutting edge: MyD88 is required for resistance to Toxoplasma gondii infection and regulates parasite-induced IL-12 production by dendritic cells. J Immunol. 2002;168(12):5997–6001. doi: 10.4049/jimmunol.168.12.5997 12055206.

14. Scharton-Kersten TM, Wynn TA, Denkers EY, Bala S, Grunvald E, Hieny S, et al. In the absence of endogenous IFN-gamma, mice develop unimpaired IL-12 responses to Toxoplasma gondii while failing to control acute infection. J Immunol. 1996;157(9):4045–54. 8892638.

15. Xuan NT, Wang X, Nishanth G, Waisman A, Borucki K, Isermann B, et al. A20 expression in dendritic cells protects mice from LPS-induced mortality. Eur J Immunol. 2014. doi: 10.1002/eji.201444795 25472594.

16. Wang X, Deckert M, Xuan NT, Nishanth G, Just S, Waisman A, et al. Astrocytic A20 ameliorates experimental autoimmune encephalomyelitis by inhibiting NF-kappaB- and STAT1-dependent chemokine production in astrocytes. Acta Neuropathol. 2013;126(5):711–24. doi: 10.1007/s00401-013-1183-9 24077734.

17. Lee EG, Boone DL, Chai S, Libby SL, Chien M, Lodolce JP, et al. Failure to regulate TNF-induced NF-kappaB and cell death responses in A20-deficient mice. Science. 2000;289(5488):2350–4. doi: 10.1126/science.289.5488.2350 11009421.

18. De Wilde K, Martens A, Lambrecht S, Jacques P, Drennan MB, Debusschere K, et al. A20 inhibition of STAT1 expression in myeloid cells: a novel endogenous regulatory mechanism preventing development of enthesitis. Ann Rheum Dis. 2017;76(3):585–92. doi: 10.1136/annrheumdis-2016-209454 27551052.

19. Johansson P, Bergmann A, Rahmann S, Wohlers I, Scholtysik R, Przekopowitz M, et al. Recurrent alterations of TNFAIP3 (A20) in T-cell large granular lymphocytic leukemia. Int J Cancer. 2016;138(1):121–4. doi: 10.1002/ijc.29697 26199174.

20. Naik SH, Proietto AI, Wilson NS, Dakic A, Schnorrer P, Fuchsberger M, et al. Cutting edge: generation of splenic CD8+ and CD8- dendritic cell equivalents in Fms-like tyrosine kinase 3 ligand bone marrow cultures. J Immunol. 2005;174(11):6592–7. doi: 10.4049/jimmunol.174.11.6592 15905497.

21. Cohen SB, Denkers EY. Impact of Toxoplasma gondii on Dendritic Cell Subset Function in the Intestinal Mucosa. J Immunol. 2015;195(6):2754–62. doi: 10.4049/jimmunol.1501137 26283477.

22. Schneider AG, Abi Abdallah DS, Butcher BA, Denkers EY. Toxoplasma gondii triggers phosphorylation and nuclear translocation of dendritic cell STAT1 while simultaneously blocking IFNgamma-induced STAT1 transcriptional activity. PLoS One. 2013;8(3):e60215. doi: 10.1371/journal.pone.0060215 23527309.

23. Yin L, Fang Z, Shen NJ, Qiu YH, Li AJ, Zhang YJ. Downregulation of A20 increases the cytotoxicity of IFN-gamma in hepatocellular carcinoma cells. Drug Des Devel Ther. 2017;11:2841–50. doi: 10.2147/DDDT.S135993 29033545.

24. Raetz M, Kibardin A, Sturge CR, Pifer R, Li H, Burstein E, et al. Cooperation of TLR12 and TLR11 in the IRF8-dependent IL-12 response to Toxoplasma gondii profilin. J Immunol. 2013;191(9):4818–27. doi: 10.4049/jimmunol.1301301 24078692.

25. Yarovinsky F, Kanzler H, Hieny S, Coffman RL, Sher A. Toll-like receptor recognition regulates immunodominance in an antimicrobial CD4+ T cell response. Immunity. 2006;25(4):655–64. doi: 10.1016/j.immuni.2006.07.015 17000122.

26. Döring Y, Soehnlein O, Drechsler M, Shagdarsuren E, Chaudhari SM, Meiler S, et al. Hematopoietic interferon regulatory factor 8-deficiency accelerates atherosclerosis in mice. 2012;32(7):1613–23. doi: 10.1161/ATVBAHA.111.236539 22556330.

27. Tosh KW, Mittereder L, Bonne-Annee S, Hieny S, Nutman TB, Singer SM, et al. The IL-12 Response of Primary Human Dendritic Cells and Monocytes to Toxoplasma gondii Is Stimulated by Phagocytosis of Live Parasites Rather Than Host Cell Invasion. J Immunol. 2016;196(1):345–56. doi: 10.4049/jimmunol.1501558 26597011.

28. Matzner N, Zemtsova IM, Nguyen TX, Duszenko M, Shumilina E, Lang F. Ion channels modulating mouse dendritic cell functions. J Immunol. 2008;181(10):6803–9. doi: 10.4049/jimmunol.181.10.6803 18981098.

29. Chen H, Hu L, Luo Z, Zhang J, Zhang C, Qiu B, et al. A20 suppresses hepatocellular carcinoma proliferation and metastasis through inhibition of Twist1 expression. Mol Cancer. 2015;14:186. doi: 10.1186/s12943-015-0454-6 26538215.

30. Longo CR, Arvelo MB, Patel VI, Daniel S, Mahiou J, Grey ST, et al. A20 protects from CD40-CD40 ligand-mediated endothelial cell activation and apoptosis. Circulation. 2003;108(9):1113–8. doi: 10.1161/01.CIR.0000083718.76889.D0 12885753.

31. Banchereau J, Briere F, Caux C, Davoust J, Lebecque S, Liu YJ, et al. Immunobiology of dendritic cells. Annu Rev Immunol. 2000;18:767–811. doi: 10.1146/annurev.immunol.18.1.767 10837075.

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