Abnormalities of dendritic cells (DCs) and STAT proteins have been reported in Crohn’s disease (CD). Studies on JAK/STAT signaling in DCs are, however, lacking in CD. We applied a flowcytometric single-cell-based phosphoepitope assay to evaluate STAT1 and STAT3 pathways in DC subsets from CD patients. In addition, circulating DC counts were determined, together with the activation-related immunophenotype. We found that IL-6- and IFN-α-induced STAT3 phosphorylation and IFN-α-induced STAT1 phosphorylation were impaired in plasmacytoid DCs (pDCs) from CD patients (P = 0.005, P = 0.013, and P = 0.006, respectively). In myeloid DCs (mDCs), IFN-α-induced STAT1 and STAT3 phosphorylation were attenuated (P<0.001 and P = 0.048, respectively), but IL-10-induced STAT3 phosphorylation was enhanced (P = 0.026). IFN-γ-induced STAT1 signaling was intact in both DC subtypes. Elevated plasma IL-6 levels were detected in CD (P = 0.004), which strongly correlated with disease activity (ρ = 0.690, P<0.001) but not with IL-6-induced STAT3 phosphorylation. The numbers of pDCs and BDCA3+ mDCs were decreased, and CD40 expression on CD1c+ mDCs was increased in CD. When elucidating the effect of IL-6 signaling on pDC function, we observed that IL-6 treatment of healthy donor pDCs affected the maturation of and modified the T-cell priming by pDCs, favoring Th2 over Th1 type of response and the expression of IL-10 in T cells. Our results implicate DC signaling in human CD. Reduced IL-6 responsiveness in pDCs, together with the attenuated IFN-α-induced signaling in both DC subtypes, may contribute to the immunological dysregulation in CD patients.
References
[1]
Brand S (2009) Crohn's disease: Th1, Th17 or both? the change of a paradigm: New immunological and genetic insights implicate Th17 cells in the pathogenesis of crohn's disease. Gut 58: 1152–1167.
[2]
MacDonald KP, Munster DJ, Clark GJ, Dzionek A, Schmitz J, et al. (2002) Characterization of human blood dendritic cell subsets. Blood 100: 4512–4520.
[3]
Lindstedt M, Lundberg K, Borrebaeck CA (2005) Gene family clustering identifies functionally associated subsets of human in vivo blood and tonsillar dendritic cells. J Immunol 175: 4839–4846.
[4]
Jonuleit H, Schmitt E, Schuler G, Knop J, Enk AH (2000) Induction of interleukin 10-producing, nonproliferating CD4(+) T cells with regulatory properties by repetitive stimulation with allogeneic immature human dendritic cells. J Exp Med 192: 1213–1222.
[5]
Ito T, Yang M, Wang YH, Lande R, Gregorio J, et al. (2007) Plasmacytoid dendritic cells prime IL-10-producing T regulatory cells by inducible costimulator ligand. J Exp Med 204: 105–115.
[6]
Kadowaki N, Antonenko S, Lau JY, Liu YJ (2000) Natural interferon alpha/beta-producing cells link innate and adaptive immunity. J Exp Med 192: 219–226.
[7]
Fonteneau JF, Gilliet M, Larsson M, Dasilva I, Munz C, et al. (2003) Activation of influenza virus-specific CD4+ and CD8+ T cells: A new role for plasmacytoid dendritic cells in adaptive immunity. Blood 101: 3520–3526.
Ochando JC, Homma C, Yang Y, Hidalgo A, Garin A, et al. (2006) Alloantigen-presenting plasmacytoid dendritic cells mediate tolerance to vascularized grafts. Nat Immunol 7: 652–662.
[10]
Jongbloed SL, Benson RA, Nickdel MB, Garside P, McInnes IB, et al. (2009) Plasmacytoid dendritic cells regulate breach of self-tolerance in autoimmune arthritis. J Immunol 182: 963–968.
[11]
Moseman EA, Liang X, Dawson AJ, Panoskaltsis-Mortari A, Krieg AM, et al. (2004) Human plasmacytoid dendritic cells activated by CpG oligodeoxynucleotides induce the generation of CD4+CD25+ regulatory T cells. J Immunol 173: 4433–4442.
[12]
Kuwana M, Kaburaki J, Wright TM, Kawakami Y, Ikeda Y (2001) Induction of antigen-specific human CD4(+) T cell anergy by peripheral blood DC2 precursors. Eur J Immunol 31: 2547–2557.
[13]
Gilliet M, Liu YJ (2002) Generation of human CD8 T regulatory cells by CD40 ligand-activated plasmacytoid dendritic cells. J Exp Med 195: 695–704.
[14]
Sakuraba A, Sato T, Kamada N, Kitazume M, Sugita A, et al. (2009) Th1/Th17 immune response is induced by mesenteric lymph node dendritic cells in crohn's disease. Gastroenterology 137: 1736–1745.
[15]
Baumgart DC, Metzke D, Schmitz J, Scheffold A, Sturm A, et al. (2005) Patients with active inflammatory bowel disease lack immature peripheral blood plasmacytoid and myeloid dendritic cells. Gut 54: 228–236.
[16]
Hart AL, Al-Hassi HO, Rigby RJ, Bell SJ, Emmanuel AV, et al. (2005) Characteristics of intestinal dendritic cells in inflammatory bowel diseases. Gastroenterology 129: 50–65.
[17]
Middel P, Raddatz D, Gunawan B, Haller F, Radzun HJ (2006) Increased number of mature dendritic cells in crohn's disease: Evidence for a chemokine mediated retention mechanism. Gut 55: 220–227.
[18]
Silva MA, Lopez CB, Riverin F, Oligny L, Menezes J, et al. (2004) Characterization and distribution of colonic dendritic cells in crohn's disease. Inflamm Bowel Dis 10: 504–512.
[19]
te Velde AA, van Kooyk Y, Braat H, Hommes DW, Dellemijn TA, et al. (2003) Increased expression of DC-SIGN+IL-12+IL-18+ and CD83+IL-12-IL-18- dendritic cell populations in the colonic mucosa of patients with crohn's disease. Eur J Immunol 33: 143–151.
[20]
Baumgart DC, Thomas S, Przesdzing I, Metzke D, Bielecki C, et al. (2009) Exaggerated inflammatory response of primary human myeloid dendritic cells to lipopolysaccharide in patients with inflammatory bowel disease. Clin Exp Immunol 157: 423–436.
[21]
Lovato P, Brender C, Agnholt J, Kelsen J, Kaltoft K, et al. (2003) Constitutive STAT3 activation in intestinal T cells from patients with crohn's disease. J Biol Chem 278: 16777–16781.
[22]
Mudter J, Weigmann B, Bartsch B, Kiesslich R, Strand D, et al. (2005) Activation pattern of signal transducers and activators of transcription (STAT) factors in inflammatory bowel diseases. Am J Gastroenterol 100: 64–72.
[23]
Musso A, Dentelli P, Carlino A, Chiusa L, Repici A, et al. (2005) Signal transducers and activators of transcription 3 signaling pathway: An essential mediator of inflammatory bowel disease and other forms of intestinal inflammation. Inflamm Bowel Dis 11: 91–98.
[24]
Schreiber S, Rosenstiel P, Hampe J, Nikolaus S, Groessner B, et al. (2002) Activation of signal transducer and activator of transcription (STAT) 1 in human chronic inflammatory bowel disease. Gut 51: 379–385.
[25]
Suzuki A, Hanada T, Mitsuyama K, Yoshida T, Kamizono S, et al. (2001) CIS3/SOCS3/SSI3 plays a negative regulatory role in STAT3 activation and intestinal inflammation. J Exp Med 193: 471–481.
[26]
Wu F, Dassopoulos T, Cope L, Maitra A, Brant SR, et al. (2007) Genome-wide gene expression differences in crohn's disease and ulcerative colitis from endoscopic pinch biopsies: Insights into distinctive pathogenesis. Inflamm Bowel Dis 13: 807–821.
[27]
Barrett JC, Hansoul S, Nicolae DL, Cho JH, Duerr RH, et al. (2008) Genome-wide association defines more than 30 distinct susceptibility loci for crohn's disease. Nat Genet 40: 955–962.
[28]
Cheng F, Wang HW, Cuenca A, Huang M, Ghansah T, et al. (2003) A critical role for Stat3 signaling in immune tolerance. Immunity 19: 425–436.
[29]
Melillo JA, Song L, Bhagat G, Blazquez AB, Plumlee CR, et al. (2010) Dendritic cell (DC)-specific targeting reveals Stat3 as a negative regulator of DC function. J Immunol 184: 2638–2645.
[30]
Jackson SH, Yu CR, Mahdi RM, Ebong S, Egwuagu CE (2004) Dendritic cell maturation requires STAT1 and is under feedback regulation by suppressors of cytokine signaling. J Immunol 172: 2307–2315.
[31]
Perez OD, Nolan GP (2002) Simultaneous measurement of multiple active kinase states using polychromatic flow cytometry. Nat Biotechnol 20: 155–162.
[32]
Daperno M, D'Haens G, Van Assche G, Baert F, Bulois P, et al. (2004) Development and validation of a new, simplified endoscopic activity score for crohn's disease: The SES-CD. Gastrointest Endosc 60: 505–512.
[33]
Honkanen J, Nieminen JK, Gao R, Luopajarvi K, Salo HM, et al. (2010) IL-17 immunity in human type 1 diabetes. J Immunol 185: 1959–1967.
[34]
Kato A, Watanabe T, Yamazaki M, Deki T, Suzuki M (2009) IL-6R distribution in normal human and cynomolgus monkey tissues. Regul Toxicol Pharmacol 53: 46–51.
[35]
Montoya M, Schiavoni G, Mattei F, Gresser I, Belardelli F, et al. (2002) Type I interferons produced by dendritic cells promote their phenotypic and functional activation. Blood 99: 3263–3271.
[36]
Kurche JS, Haluszczak C, McWilliams JA, Sanchez PJ, Kedl RM (2012) Type I IFN-dependent T cell activation is mediated by IFN-dependent dendritic cell OX40 ligand expression and is independent of T cell IFNR expression. J Immunol 188: 585–593.
[37]
Le Bon A, Etchart N, Rossmann C, Ashton M, Hou S, et al. (2003) Cross-priming of CD8+ T cells stimulated by virus-induced type I interferon. Nat Immunol 4: 1009–1015.
[38]
Ito T, Amakawa R, Inaba M, Ikehara S, Inaba K, et al. (2001) Differential regulation of human blood dendritic cell subsets by IFNs. J Immunol 166: 2961–2969.
[39]
Mahida YR, Kurlac L, Gallagher A, Hawkey CJ (1991) High circulating concentrations of interleukin-6 in active crohn's disease but not ulcerative colitis. Gut 32: 1531–1534.
[40]
Nancey S, Hamzaoui N, Moussata D, Graber I, Bienvenu J, et al. (2008) Serum interleukin-6, soluble interleukin-6 receptor and crohn's disease activity. Dig Dis Sci 53: 242–247.
[41]
Baumgart DC, Metzke D, Guckelberger O, Pascher A, Grotzinger C, et al. (2011) Aberrant plasmacytoid dendritic cell distribution and function in patients with crohn's disease and ulcerative colitis. Clin Exp Immunol 166: 46–54.
[42]
Lunz JG,3rd, Specht SM, Murase N, Isse K, Demetris AJ (2007) Gut-derived commensal bacterial products inhibit liver dendritic cell maturation by stimulating hepatic interleukin-6/signal transducer and activator of transcription 3 activity. Hepatology 46: 1946–1959.
Steinbrink K, Wolfl M, Jonuleit H, Knop J, Enk AH (1997) Induction of tolerance by IL-10-treated dendritic cells. J Immunol 159: 4772–4780.
[45]
Willson TA, Jurickova I, Collins M, Denson LA (2013) Deletion of intestinal epithelial cell STAT3 promotes T-lymphocyte STAT3 activation and chronic colitis following acute dextran sodium sulfate injury in mice. Inflamm Bowel Dis 19: 512–525.
[46]
Takeda K, Clausen BE, Kaisho T, Tsujimura T, Terada N, et al. (1999) Enhanced Th1 activity and development of chronic enterocolitis in mice devoid of Stat3 in macrophages and neutrophils. Immunity 10: 39–49.
[47]
Mathur AN, Chang HC, Zisoulis DG, Stritesky GL, Yu Q, et al. (2007) Stat3 and Stat4 direct development of IL-17-secreting th cells. J Immunol 178: 4901–4907.
[48]
Fujino S, Andoh A, Bamba S, Ogawa A, Hata K, et al. (2003) Increased expression of interleukin 17 in inflammatory bowel disease. Gut 52: 65–70.
[49]
Holtta V, Klemetti P, Sipponen T, Westerholm-Ormio M, Kociubinski G, et al. (2008) IL-23/IL-17 immunity as a hallmark of crohn's disease. Inflamm Bowel Dis 14: 1175–1184.
[50]
Willson TA, Kuhn BR, Jurickova I, Gerad S, Moon D, et al. (2012) STAT3 genotypic variation and cellular STAT3 activation and colon leukocyte recruitment in pediatric crohn disease. J Pediatr Gastroenterol Nutr 55: 32–43.
