Abstract
Background
Tolerance-inducing DC are considered to be less mature than immunogenic DC, but the conditions promoting a less mature DC phenotype are not clear. We have previously shown that lipopolysaccharide (LPS) can have differential effects on DC function depending on the timing of DC exposure to LPS. Here, we show that early LPS-activated bone marrow derived DC (early DC, eDC), when administered subcutaneously to mice in vivo, promote tolerance to EAU induced via immunisation with interphotoreceptor retinol binding protein (IRBP) peptide 161–180. The effect correlates with the failure of eDC to secrete IL-12, and appears to be mediated in part via expansion of naturally occurring CD4+CD25+ T regulatory cells (Tregs), which also mediate suppression of EAU on adoptive transfer to naive mice followed by immunization with autoantigen.
Methods
Immature DC were prepared from BMDC cultures. Early DC (eDC) and late DC (lDC) for tolerance experiments were obtained by differential timing of LPS addition and their cytokine secretion profile was analyzed. eDC and lDC were subcutaneously injected into mice. From the dLN CD4+ CD25+ GITR+ T regulatory cells found to express FoxP3 were isolated and transferred into mice prior to immunisation with IRBP. The immune response was scored by histopathology. Tregs were characterized in vitro by intracellular staining, cytokine secretion assay and transwell experiments.
Results
eDC secrete IL-10 but no IL-12 or IFNγ. When injected subcutaneously into naive mice, they expand the population of CD4+CD25+high GITR+ T cells expressing FoxP3 in the dLN, thus increasing the total number of IL-10 producing cells. eDC induced Tregs inhibit CD4+CD25− T effector cell proliferation by a contact dependent process, and both eDC and Tregs suppress retinal damage when adoptively transferred.
Conclusions
We suggest that DC maturation may be necessary for both tolerance and immunity, but differential levels of activation and/or cytokine production direct the outcome of DC-T cell interaction and this is determined by IL-12 production. T regulatory cells induced in vivo by contact with eDC are able to suppress disease in the EAU model by adoptive transfer.
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References
Chan CC, Caspi RR, Ni M, Leake WC, Wiggert B, Chader GC, Nussenblatt RB (1990) Pathology of experimental autoimmune uveoretinitis in mice. J Autoimmun 3:247–255
Chorny A, Gonzales-Rey E, Fernandez-Martin A, Pozo D, Ganea D, Delgado M (2005) Vasoactive intestinal peptide induces regulatory dendritic cells with therapeutic effects on autoimmune disorders. Proc Natl Acad Sci USA 102:13562–13567
Diehl S et al. (2000) Inhibition of Th1 differentiation by IL-6 is mediated by SOCS1. Immunity 13:805–815
Dodge IL, Carr MW, Cernadas M, Brenner MB (2003) IL-6 production by pulmonary dendritic cells impedes Th1 immune responses. J Immunol 170:4457–4464
Ferguson TA, Kazama H (2005) Signals from dying cells: tolerance induction by the dendritic cell. Immunol Res 32:99–108
Fontenot JD, Gavin MA, Rudensky AY (2003) Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat Immunol 4:330–336
Forrester JV, McMenamin PG (1999) Immunopathogenic mechanisms in intraocular inflammation. Chem Immunol 73:159–185
Gur H et al. (2005) Immune regulatory activity of CD34+ progenitor cells: evidence for a deletion based mechanism mediated by TNF-α. Blood 105:2585–2593
Hegde S, Pahne J, Smola-Hess S (2004) Novel immunosuppressive properties of interleukin-6 in dendritic cells: inhibition of NF-kappaB binding activity and CCR7 expression. FASEB J 18:1439–1441
Hori S, Nomura T, Sakaguchi S (2003) Control of regulatory T cell development by the transcription factor Foxp3. Science 299:1057–1066
Horng T, Barton GM, Flavell RA, Medzhitov R (2002) The adaptor molecule TIRAP provides signalling specificity for Toll-like receptors. Nature 420:329–333
Inaba K, Inaba M, Witmer-Pack M, Hatchcock K, Hodes R, Steinman RM (1995) Expression of B7 costimulator molecules on mouse dendritic cells. Adv Exp Med Biol 378:65–70
Inaba K et al. (1992) Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med 176:1693–1702
Jiang HR, Muckersie E, Robertson M, Forrester JV (2003) Antigen-specific inhibition of experimental autoimmune uveoretinitis by bone marrow-derived immature dendritic cells. Invest Ophthalmol Vis Sci 44:1598–1607
Jiang HR, Muckersie E, Robertson M, Xu H, Liversidge J, Forrester JV (2002) Secretion of interleukin-10 or interleukin-12 by LPS-activated dendritic cells is critically dependent on time of stimulus relative to initiation of purified DC culture. J Leukoc Biol 72:978–985
Jonuleit H, Schmitt E, Kakirman H, Stassen M, Knop J, Enk AH (2002) Infectious tolerance: human CD25(+) regulatory T cells convey suppressor activity to conventional CD4(+) T helper cells. J Exp Med 196:255–260
Kaisho T, Takeuchi O, Kawai T, Hoshino K, Akira S (2001) Endotoxin-induced maturation of MyD88-deficient dendritic cells. J Immunol 166:5688–5694
Kawai T, Adachi O, Ogawa T, Takeda K, Akira S (1999) Unresponsiveness of MyD88 deficient mice to endotoxin. Immunity 11:115–122
Kleindienst P et al. (2005) Simultaneous induction of CD4 T cell tolerance and CD8 T cell immunity by semimature dendritic cells. J Immunol 174:3941–3947
La Flamme AC, MacDonald AS, Pearce EJ (2000) Role of IL-6 in directing the initial immune response to schistosome eggs. J Immunol 164:2419–2426
Lutz MB (2004) IL-3 in dendritic cell development and function: a comparison with GM CSF and IL-4. Immunobiology 209:79–87
Lutz MB, Kukutsch NA, Menges M, Rossner S, Schuler G (2000) Culture of bone marrow cells in GM-CSF plus high doses of lipopolysaccharide generates exclusively immature dendritic cells which induce alloantigen-specific CD4 T cell anergy in vitro. Eur J Immunol 30:1048–1052
Mahnke K, Qian Y, Knop J, Enk AH (2003) Induction of CD4+/CD25+ regulatory T cells by targeting of antigens to immature dendritic cells. Blood 101:4862–4869
Mahnke K, Enk AH (2005) Dendritic cells: key cells for the induction of regulatory T cells? Curr Top Microbiol Immunol 293:133–150
McGuirk P, McCann C, Mills KH (2002) Pathogen-specific T regulatory 1 cells induced in the respiratory tract by a bacterial molecule that stimulates interleukin 10 production by dendritic cells: a novel strategy for evasion of protective T helper type 1 responses by Bordetella pertussis. J Exp Med 195:221–231
McHugh RS, Shevach EM (2002) The role of suppressor T cells in regulation of immune responses. J Allergy Clin Immunol 110:693–702
Menges M et al. (2002) Repetitive injections of dendritic cells matured with tumor necrosis factor alpha induce antigen-specific protection of mice from autoimmunity. J Exp Med 195:15–21
Pasare C, Medzhitov R (2004) Toll-dependent control mechanisms of CD4 T cell activation. Immunity 21:733–741
Pasare C, Medzhitov R (2003) Toll-like receptors: balancing host resistance with immune tolerance. Curr Opin Immunol 15:677–682
Pasare C, Medzhitov R (2003) Toll pathway-dependent blockade of CD4+CD25+ T cell mediated suppression by dendritic cells. Science 299:1033–1036
Shevach EM (2002) CD4+ CD25+ suppressor T cells: more questions than answers. Nat Rev Immunol 2:389–400
Silver PB, Rizzo, LV, Chan CC, Donoso LA, Wiggert, B, Caspi RR (1995) Identification of a major pathogenic epitope in the human IRBP molecule recognized by mice of the H-2r haplotype. Invest Ophthalmol Vis Sci 36:946–954
Stassen M et al. (2004) Differential regulatory capacity of CD25+ T regulatory cells and preactivated CD25+ T regulatory cells on development, functional activation, and proliferation of Th2 cells. J Immunol 173:267–274
Takenaka H et al. (1997) Regulation of T cell-dependent and -independent IL-12 production by the three Th2-type cytokines IL-10, IL-6, and IL-4. J Leukoc Biol 61:80–87
Tan JK, O’Neill HC (2005) Maturation requirements for dendritic cells in T cell stimulation leading to tolerance versus immunity. J Leukoc Biol 78:319–324
Verginis P, Li HS, Carayanniotis G (2005) Tolerogenic semimature dendritic cells suppress experimental autoimmune thyroiditis by activation of thyroglobulin-specific CD4+CD25+ T cells. J Immunol 174:7433–7439
Yamamoto M et al. (2003) Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway. Science 301:640–643
Yamamoto M et al. (2003) TRAM is specifically involved in the Toll-like receptor 4 ediated MyD88-independent signaling pathway. Nat Immunol 4:1144–1150
Yao Y, Li W, Kaplan MH, Chang CH (2005) Interleukin (IL)-4 inhibits IL-10 to promote IL-12 production by dendritic cells. J Exp Med 201:1899–1903
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Siepmann, K., Biester, S., Plšková, J. et al. CD4+CD25+ T regulatory cells induced by LPS-activated bone marrow dendritic cells suppress experimental autoimmune uveoretinitis in vivo. Graefe's Arch Clin Exp Ophthalmol 245, 221–229 (2007). https://doi.org/10.1007/s00417-006-0356-9
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DOI: https://doi.org/10.1007/s00417-006-0356-9