Interferon regulatory factor 5 activation in monocytes of systemic lupus erythematosus patients is triggered by circulating autoantigens independent of type I interferons
Rivka C. Stone
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
Search for more papers by this authorDi Feng
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
Search for more papers by this authorJing Deng
University of Medicine and Dentistry of New Jersey, Newark
Search for more papers by this authorSukhwinder Singh
University of Medicine and Dentistry of New Jersey, Newark
Search for more papers by this authorLisong Yang
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
Search for more papers by this authorPatricia Fitzgerald-Bocarsly
University of Medicine and Dentistry of New Jersey, Newark
Search for more papers by this authorCorresponding Author
Betsy J. Barnes
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
New Jersey Medical School–University Hospital Cancer Center, G1224, 205 South Orange Avenue, Newark, NJ 07103Search for more papers by this authorRivka C. Stone
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
Search for more papers by this authorDi Feng
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
Search for more papers by this authorJing Deng
University of Medicine and Dentistry of New Jersey, Newark
Search for more papers by this authorSukhwinder Singh
University of Medicine and Dentistry of New Jersey, Newark
Search for more papers by this authorLisong Yang
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
Search for more papers by this authorPatricia Fitzgerald-Bocarsly
University of Medicine and Dentistry of New Jersey, Newark
Search for more papers by this authorCorresponding Author
Betsy J. Barnes
University of Medicine and Dentistry of New Jersey and New Jersey Medical School–University Hospital Cancer Center, Newark
New Jersey Medical School–University Hospital Cancer Center, G1224, 205 South Orange Avenue, Newark, NJ 07103Search for more papers by this authorAbstract
Objective
Genetic variants of interferon regulatory factor 5 (IRF-5) are associated with susceptibility to systemic lupus erythematosus (SLE). IRF-5 regulates the expression of proinflammatory cytokines and type I interferons (IFNs) believed to be involved in the pathogenesis of SLE. The aim of this study was to determine the activation status of IRF-5 by assessing its nuclear localization in the immune cells of SLE patients and healthy donors, and to identify SLE-associated triggers of IRF-5 activation.
Methods
IRF-5 nuclear localization in subpopulations of peripheral blood mononuclear cells from 14 genotyped SLE patients and 11 healthy controls was assessed using imaging flow cytometry. The activation and function of IRF-5 were examined after ex vivo stimulation of healthy donor monocytes with SLE serum or components of SLE serum. Cellular localization was determined by ImageStream flow cytometry, and cytokine expression was analyzed by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay.
Results
IRF-5 was activated in a cell type–specific manner; monocytes from SLE patients had constitutively elevated levels of nuclear IRF-5, as compared to natural killer cells and T cells. SLE serum was identified as a trigger for IRF-5 nuclear accumulation; however, neither IFNα nor SLE immune complexes could induce nuclear localization. Instead, autoantigens composed of apoptotic/necrotic material triggered IRF-5 nuclear accumulation in monocytes. Production of the cytokines IFNα, tumor necrosis factor α, and interleukin-6 in monocytes stimulated with SLE serum or autoantigens was distinct, yet showed a correlation with the kinetics of IRF-5 nuclear localization.
Conclusion
This study provides the first formal proof that IRF-5 activation is altered in the monocytes of SLE patients, which can be attributed, in part, to the SLE blood environment.
REFERENCES
- 1 Ronnblom L, Alm GV, Eloranta ML. The type I interferon system in the development of lupus. Semin Immunol 2011; 23: 113–21.
- 2 Graham RR, Kyogoku C, Sigurdsson S, Vlasova IA, Davies LR, Baechler EC, et al. Three functional variants of IFN regulatory factor 5 (IRF5) define risk and protective haplotypes for human lupus. Proc Natl Acad Sci U S A 2007; 104: 6758–63.
- 3 Kelly JA, Kelley JM, Kaufman KM, Kilpatrick J, Bruner GR, Merrill JT, et al. Interferon regulatory factor-5 is genetically associated with systemic lupus erythematosus in African Americans. Genes Immun 2008; 9: 187–94.
- 4 Shin HD, Sung YK, Choi CB, Lee SO, Lee HW, Bae SC. Replication of the genetic effects of IFN regulatory factor 5 (IRF5) on systemic lupus erythematosus in a Korean population. Arthritis Res Ther 2007; 9: R32.
- 5 Sigurdsson S, Goring HH, Kristjansdottir G, Milani L, Nordmark G, Sandling JK, et al. Comprehensive evaluation of the genetic variants of interferon regulatory factor 5 (IRF5) reveals a novel 5 bp length polymorphism as strong risk factor for systemic lupus erythematosus. Hum Mol Genet 2008; 17: 872–81.
- 6 Barnes BJ, Kellum MJ, Field AE, Pitha PM. Multiple regulatory domains of IRF-5 control activation, cellular localization, and induction of chemokines that mediate recruitment of T lymphocytes. Mol Cell Biol 2002; 22: 5721–40.
- 7 Barnes BJ, Moore PA, Pitha PM. Virus-specific activation of a novel interferon regulatory factor, IRF-5, results in the induction of distinct interferon α genes. J Biol Chem 2001; 276: 23382–90.
- 8 Barnes BJ, Richards J, Mancl M, Hanash S, Beretta L, Pitha PM. Global and distinct targets of IRF-5 and IRF-7 during innate response to viral infection. J Biol Chem 2004; 279: 45194–207.
- 9 Takaoka A, Yanai H, Kondo S, Duncan G, Negishi H, Mizutani T, et al. Integral role of IRF-5 in the gene induction programme activated by Toll-like receptors. Nature 2005; 434: 243–9.
- 10 Barnes BJ, Kellum MJ, Pinder KE, Frisancho JA, Pitha PM. Interferon regulatory factor 5, a novel mediator of cell cycle arrest and cell death. Cancer Res 2003; 63: 6424–31.
- 11 Yanai H, Chen HM, Inuzuka T, Kondo S, Mak TW, Takaoka A, et al. Role of IFN regulatory factor 5 transcription factor in antiviral immunity and tumor suppression. Proc Natl Acad Sci U S A 2007; 104: 3402–7.
- 12 Feng D, Stone RC, Eloranta ML, Sangster-Guity N, Nordmark G, Sigurdsson S, et al. Genetic variants and disease-associated factors contribute to enhanced interferon regulatory factor 5 expression in blood cells of patients with systemic lupus erythematosus. Arthritis Rheum 2010; 62: 562–73.
- 13 Mancl ME, Hu G, Sangster-Guity N, Olshalsky SL, Hoops K, Fitzgerald-Bocarsly P, et al. Two discrete promoters regulate the alternatively spliced human interferon regulatory factor-5 isoforms: multiple isoforms with distinct cell type-specific expression, localization, regulation, and function. J Biol Chem 2005; 280: 21078–90.
- 14 Kyttaris VC, Katsiari CG, Juang YT, Tsokos GC. New insights into the pathogenesis of systemic lupus erythematosus. Curr Rheumatol Rep 2005; 7: 469–75.
- 15 Auger MJ, Ross JA. The biology of macrophage. In: CE Lewis, JO McGee, editors. The macrophage. Oxford: Oxford University Press; 2008. p. 2–74.
- 16 Katsiari CG, Liossis SN, Sfikakis PP. The pathophysiologic role of monocytes and macrophages in systemic lupus erythematosus: a reappraisal. Semin Arthritis Rheum 2010; 39: 491–503.
- 17 Unanue ER. The regulation of lymphocyte functions by the macrophage. Immunol Rev 1978; 40: 227–55.
- 18
Baumann I,
Kolowos W,
Voll RE,
Manger B,
Gaipl U,
Neuhuber WL, et al.
Impaired uptake of apoptotic cells into tingible body macrophages in germinal centers of patients with systemic lupus erythematosus.
Arthritis Rheum
2002;
46:
191–201.
10.1002/1529-0131(200201)46:1<191::AID-ART10027>3.0.CO;2-K CAS PubMed Web of Science® Google Scholar
- 19 Steinbach F, Henke F, Krause B, Thiele B, Burmester GR, Hiepe F. Monocytes from systemic lupus erythematous patients are severely altered in phenotype and lineage flexibility. Ann Rheum Dis 2000; 59: 283–8.
- 20 Hu G, Barnes BJ. IRF-5 is a mediator of the death receptor-induced apoptotic signaling pathway. J Biol Chem 2009; 284: 2767–77.
- 21 Hu G, Mancl ME, Barnes BJ. Signaling through IFN regulatory factor-5 sensitizes p53-deficient tumors to DNA damage-induced apoptosis and cell death. Cancer Res 2005; 65: 7403–12.
- 22 Schoenemeyer A, Barnes BJ, Mancl ME, Latz E, Goutagny N, Pitha PM, et al. The interferon regulatory factor, IRF5, is a central mediator of Toll-like receptor 7 signaling. J Biol Chem 2005; 280: 17005–12.
- 23 Tan EM, Cohen AS, Fries JF, Masi AT, McShane DJ, Rothfield NF, et al. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 1982; 25: 1271–7.
- 24 Gladman DD, Ibanez D, Urowitz MB. Systemic Lupus Erythematosus Disease Activity Index 2000. J Rheumatol 2002; 29: 288–91.
- 25 Danis B, George TC, Goriely S, Dutta B, Renneson J, Gatto L, et al. Interferon regulatory factor 7-mediated responses are defective in cord blood plasmacytoid dendritic cells. Eur J Immunol 2008; 38: 507–17.
- 26 Fanning SL, George TC, Feng D, Feldman SB, Megjugorac NJ, Izaguirre AG, et al. Receptor cross-linking on human plasmacytoid dendritic cells leads to the regulation of IFN-α production. J Immunol 2006; 177: 5829–39.
- 27 George TC, Fanning SL, Fitzgerald-Bocarsly P, Medeiros RB, Highfill S, Shimizu Y, et al. Quantitative measurement of nuclear translocation events using similarity analysis of multispectral cellular images obtained in flow. J Immunol Methods 2006; 311: 117–29.
- 28 Wen F, Ellingson SM, Kyogoku C, Peterson EJ, Gaffney PM. Exon 6 variants carried on systemic lupus erythematosus (SLE) risk haplotypes modulate IRF5 function. Autoimmunity 2011; 44: 82–9.
- 29 Calzada-Nova G, Schnitzlein WM, Husmann RJ, Zuckermann FA. North American porcine reproductive and respiratory syndrome viruses inhibit type I interferon production by plasmacytoid dendritic cells. J Virol 2011; 85: 2703–13.
- 30 Vallin H, Blomberg S, Alm GV, Cederblad B, Ronnblom L. Patients with systemic lupus erythematosus (SLE) have a circulating inducer of interferon-alpha (IFN-α) production acting on leucocytes resembling immature dendritic cells. Clin Exp Immunol 1999; 115: 196–202.
- 31 Hua J, Kirou K, Lee C, Crow MK. Functional assay of type I interferon in systemic lupus erythematosus plasma and association with anti–RNA binding protein autoantibodies. Arthritis Rheum 2006; 54: 1906–16.
- 32 Lovgren T, Eloranta ML, Bave U, Alm GV, Ronnblom L. Induction of interferon-α production in plasmacytoid dendritic cells by immune complexes containing nucleic acid released by necrotic or late apoptotic cells and lupus IgG. Arthritis Rheum 2004; 50: 1861–72.
- 33 Li Y, Lee PY, Reeves WH. Monocyte and macrophage abnormalities in systemic lupus erythematosus. Arch Immunol Ther Exp (Warsz) 2010; 58: 355–64.
- 34 Cairns AP, Crockard AD, McConnell JR, Courtney PA, Bell AL. Reduced expression of CD44 on monocytes and neutrophils in systemic lupus erythematosus: relations with apoptotic neutrophils and disease activity. Ann Rheum Dis 2001; 60: 950–5.
- 35 Gaipl US, Voll RE, Sheriff A, Franz S, Kalden JR, Herrmann M. Impaired clearance of dying cells in systemic lupus erythematosus. Autoimmun Rev 2005; 4: 189–94.
- 36 Baechler EC, Batliwalla FM, Karypis G, Gaffney PM, Ortmann WA, Espe KJ, et al. Interferon-inducible gene expression signature in peripheral blood cells of patients with severe lupus. Proc Natl Acad Sci U S A 2003; 100: 2610–5.
- 37 Ho V, McLean A, Terry S. Severe systemic lupus erythematosus induced by antiviral treatment for hepatitis C. J Clin Rheumatol 2008; 14: 166–8.
- 38 Hooks JJ, Moutsopoulos HM, Geis SA, Stahl NI, Decker JL, Notkins AL. Immune interferon in the circulation of patients with autoimmune disease. N Engl J Med 1979; 301: 5–8.
- 39 Munoz LE, van Bavel C, Franz S, Berden J, Herrmann M, van der Vlag J. Apoptosis in the pathogenesis of systemic lupus erythematosus. Lupus 2008; 17: 371–5.
- 40 Rumore PM, Steinman CR. Endogenous circulating DNA in systemic lupus erythematosus: occurrence as multimeric complexes bound to histone. J Clin Invest 1990; 86: 69–74.
- 41 Ronnblom L. Potential role of IFNα in adult lupus. Arthritis Res Ther 2010; 12 Suppl 1: S3.
- 42 Hornung V, Rothenfusser S, Britsch S, Krug A, Jahrsdorfer B, Giese T, et al. Quantitative expression of Toll-like receptor 1-10 mRNA in cellular subsets of human peripheral blood mononuclear cells and sensitivity to CpG oligodeoxynucleotides. J Immunol 2002; 168: 4531–7.
- 43 Fu Y, Comella N, Tognazzi K, Brown LF, Dvorak HF, Kocher O. Cloning of DLM-1, a novel gene that is up-regulated in activated macrophages, using RNA differential display. Gene 1999; 240: 157–63.
- 44 Takaoka A, Wang ZC, Choi MK, Yanai H, Negishi H, Ban T, et al. DAI (DLM-1/ZBP1) cytosolic DNA sensor and an activator of innate immune response. Nature 2007; 448: 501–5.
- 45 Sharma S, Fitzgerald KA. Innate immune sensing of DNA. PLoS Pathog 2011; 7: e1001310.
- 46 Green DR, Ferguson T, Zitvogel L, Kroemer G. Immunogenic and tolerogenic cell death. Nat Rev Immunol 2009; 9: 353–63.
- 47 Debets JM, van der Linden CJ, Dieteren IE, Leeuwenberg JF, Buurman WA. Fc-receptor cross-linking induces rapid secretion of tumor necrosis factor (cachectin) by human peripheral blood monocytes. J Immunol 1988; 141: 1197–201.
