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Repetitive injections of dendritic cells matured with tumor necrosis factor alpha induce antigen-specific protection of mice from autoimmunity.

Menges M, Rössner S, Voigtländer C, Schindler H, Kukutsch NA, Bogdan C, Erb K, Schuler G, Lutz MB - J. Exp. Med. (2002)

Bottom Line: Three consecutive injections of peptide-pulsed TNF/DCs derived from wild-type led to the induction of peptide-specific predominantly interleukin (IL)-10-producing CD4(+) T cells and complete protection from EAE.Notably, the protection was peptide specific, as TNF/DCs pulsed with unrelated peptide could not prevent EAE.In conclusion, this study describes that stimulation by TNF-alpha results in incompletely matured DCs (semi-mature DCs) which induce peptide-specific IL-10-producing T cells in vivo and prevent EAE.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Erlangen, Erlangen 91052, Germany.

ABSTRACT
Mature dendritic cells (DCs) are believed to induce T cell immunity, whereas immature DCs induce T cell tolerance. Here we describe that injections of DCs matured with tumor necrosis factor (TNF)-alpha (TNF/DCs) induce antigen-specific protection from experimental autoimmune encephalomyelitis (EAE) in mice. Maturation by TNF-alpha induced high levels of major histocompatibility complex class II and costimulatory molecules on DCs, but they remained weak producers of proinflammatory cytokines. One injection of such TNF/DCs pulsed with auto-antigenic peptide ameliorated the disease score of EAE. This could not be observed with immature DCs or DCs matured with lipopolysaccharide (LPS) plus anti-CD40. Three consecutive injections of peptide-pulsed TNF/DCs derived from wild-type led to the induction of peptide-specific predominantly interleukin (IL)-10-producing CD4(+) T cells and complete protection from EAE. Blocking of IL-10 in vivo could only partially restore the susceptibility to EAE, suggesting an important but not exclusive role of IL-10 for EAE prevention. Notably, the protection was peptide specific, as TNF/DCs pulsed with unrelated peptide could not prevent EAE. In conclusion, this study describes that stimulation by TNF-alpha results in incompletely matured DCs (semi-mature DCs) which induce peptide-specific IL-10-producing T cells in vivo and prevent EAE.

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Repetitive injections of TNF/DCs protect mice from EAE DCs were simultaneously pulsed with MOG peptide and treated with TNF-α for 4 h (MOG/TNF/DC), washed in PBS, and 2.5 × 106 cells were injected once (1×) or three times (3×) intravenously into 3–4 C57BL/6 mice per group. Control mice were left without DC injections (untreated). 3 d after the last/only DC injection EAE was induced and the mice observed for paralysis. (a) Injection of immature DCs, generated in the presence of IL-10, pulsed with MOG peptide, and injected 3 d before EAE induction, were not protective. (b) DCs matured with LPS plus anti-CD40 and pulsed with MOG peptide and injected before EAE induction were not protective. (c) Single injections of MOG pulsed and TNF-α matured DCs, given 7 d before EAE induction, could ameliorate the disease. (d) Three injections of MOG-pulsed TNF/DC but not unpulsed TNF/DC given at days −7, −5, and −3 before EAE induction completely protected mice from EAE. (e) OVA/TNF/DCs could not protect from EAE. As a peptide specificity control, three injections of OVA-pulsed TNF/DC were given at days −7, −5, and −3 before EAE induction or mice were left untreated. OVA peptide was additionally emulsified together with the MOG peptide in CFA at day 0 in this experiment. The data for a–d are each representative of three independent experiments with similar results.
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fig2: Repetitive injections of TNF/DCs protect mice from EAE DCs were simultaneously pulsed with MOG peptide and treated with TNF-α for 4 h (MOG/TNF/DC), washed in PBS, and 2.5 × 106 cells were injected once (1×) or three times (3×) intravenously into 3–4 C57BL/6 mice per group. Control mice were left without DC injections (untreated). 3 d after the last/only DC injection EAE was induced and the mice observed for paralysis. (a) Injection of immature DCs, generated in the presence of IL-10, pulsed with MOG peptide, and injected 3 d before EAE induction, were not protective. (b) DCs matured with LPS plus anti-CD40 and pulsed with MOG peptide and injected before EAE induction were not protective. (c) Single injections of MOG pulsed and TNF-α matured DCs, given 7 d before EAE induction, could ameliorate the disease. (d) Three injections of MOG-pulsed TNF/DC but not unpulsed TNF/DC given at days −7, −5, and −3 before EAE induction completely protected mice from EAE. (e) OVA/TNF/DCs could not protect from EAE. As a peptide specificity control, three injections of OVA-pulsed TNF/DC were given at days −7, −5, and −3 before EAE induction or mice were left untreated. OVA peptide was additionally emulsified together with the MOG peptide in CFA at day 0 in this experiment. The data for a–d are each representative of three independent experiments with similar results.

Mentions: Immature DCs, generated by various methods, have been shown to induce T cell anergy or regulatory T cells and that they can act in tolerogenic fashion in mice (8, 21, 22), rat (14), and human (10). Therefore, we generated immature DCs by the continuous presence of IL-10 (19), pulsed them with the auto-antigenic MOG peptide 35–55, and injected them intravenously into mice 3 d before EAE induction. However, the EAE score was not influenced by these immature DC (Fig. 2 a) or MOG-pulsed DCs matured with LPS plus anti-CD40 (Fig. 2 b). Also three injections of MOG/LPS/CD40/DCs did not prevent EAE (not shown). Surprisingly, a single injection of MOG-pulsed TNF/DCs given 3, 5 (not shown), or 7 d before EAE induction ameliorated the course of the disease (Fig. 2 c), and three injections led to complete prevention of the disease. This prevention from EAE was dependent on the loading of TNF/DCs with MOG peptide and was not observed by unloaded TNF/DCs (Fig. 2 d) or when TNF/DCs were pulsed with irrelevant OVA peptide.


Repetitive injections of dendritic cells matured with tumor necrosis factor alpha induce antigen-specific protection of mice from autoimmunity.

Menges M, Rössner S, Voigtländer C, Schindler H, Kukutsch NA, Bogdan C, Erb K, Schuler G, Lutz MB - J. Exp. Med. (2002)

Repetitive injections of TNF/DCs protect mice from EAE DCs were simultaneously pulsed with MOG peptide and treated with TNF-α for 4 h (MOG/TNF/DC), washed in PBS, and 2.5 × 106 cells were injected once (1×) or three times (3×) intravenously into 3–4 C57BL/6 mice per group. Control mice were left without DC injections (untreated). 3 d after the last/only DC injection EAE was induced and the mice observed for paralysis. (a) Injection of immature DCs, generated in the presence of IL-10, pulsed with MOG peptide, and injected 3 d before EAE induction, were not protective. (b) DCs matured with LPS plus anti-CD40 and pulsed with MOG peptide and injected before EAE induction were not protective. (c) Single injections of MOG pulsed and TNF-α matured DCs, given 7 d before EAE induction, could ameliorate the disease. (d) Three injections of MOG-pulsed TNF/DC but not unpulsed TNF/DC given at days −7, −5, and −3 before EAE induction completely protected mice from EAE. (e) OVA/TNF/DCs could not protect from EAE. As a peptide specificity control, three injections of OVA-pulsed TNF/DC were given at days −7, −5, and −3 before EAE induction or mice were left untreated. OVA peptide was additionally emulsified together with the MOG peptide in CFA at day 0 in this experiment. The data for a–d are each representative of three independent experiments with similar results.
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Related In: Results  -  Collection

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fig2: Repetitive injections of TNF/DCs protect mice from EAE DCs were simultaneously pulsed with MOG peptide and treated with TNF-α for 4 h (MOG/TNF/DC), washed in PBS, and 2.5 × 106 cells were injected once (1×) or three times (3×) intravenously into 3–4 C57BL/6 mice per group. Control mice were left without DC injections (untreated). 3 d after the last/only DC injection EAE was induced and the mice observed for paralysis. (a) Injection of immature DCs, generated in the presence of IL-10, pulsed with MOG peptide, and injected 3 d before EAE induction, were not protective. (b) DCs matured with LPS plus anti-CD40 and pulsed with MOG peptide and injected before EAE induction were not protective. (c) Single injections of MOG pulsed and TNF-α matured DCs, given 7 d before EAE induction, could ameliorate the disease. (d) Three injections of MOG-pulsed TNF/DC but not unpulsed TNF/DC given at days −7, −5, and −3 before EAE induction completely protected mice from EAE. (e) OVA/TNF/DCs could not protect from EAE. As a peptide specificity control, three injections of OVA-pulsed TNF/DC were given at days −7, −5, and −3 before EAE induction or mice were left untreated. OVA peptide was additionally emulsified together with the MOG peptide in CFA at day 0 in this experiment. The data for a–d are each representative of three independent experiments with similar results.
Mentions: Immature DCs, generated by various methods, have been shown to induce T cell anergy or regulatory T cells and that they can act in tolerogenic fashion in mice (8, 21, 22), rat (14), and human (10). Therefore, we generated immature DCs by the continuous presence of IL-10 (19), pulsed them with the auto-antigenic MOG peptide 35–55, and injected them intravenously into mice 3 d before EAE induction. However, the EAE score was not influenced by these immature DC (Fig. 2 a) or MOG-pulsed DCs matured with LPS plus anti-CD40 (Fig. 2 b). Also three injections of MOG/LPS/CD40/DCs did not prevent EAE (not shown). Surprisingly, a single injection of MOG-pulsed TNF/DCs given 3, 5 (not shown), or 7 d before EAE induction ameliorated the course of the disease (Fig. 2 c), and three injections led to complete prevention of the disease. This prevention from EAE was dependent on the loading of TNF/DCs with MOG peptide and was not observed by unloaded TNF/DCs (Fig. 2 d) or when TNF/DCs were pulsed with irrelevant OVA peptide.

Bottom Line: Three consecutive injections of peptide-pulsed TNF/DCs derived from wild-type led to the induction of peptide-specific predominantly interleukin (IL)-10-producing CD4(+) T cells and complete protection from EAE.Notably, the protection was peptide specific, as TNF/DCs pulsed with unrelated peptide could not prevent EAE.In conclusion, this study describes that stimulation by TNF-alpha results in incompletely matured DCs (semi-mature DCs) which induce peptide-specific IL-10-producing T cells in vivo and prevent EAE.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Erlangen, Erlangen 91052, Germany.

ABSTRACT
Mature dendritic cells (DCs) are believed to induce T cell immunity, whereas immature DCs induce T cell tolerance. Here we describe that injections of DCs matured with tumor necrosis factor (TNF)-alpha (TNF/DCs) induce antigen-specific protection from experimental autoimmune encephalomyelitis (EAE) in mice. Maturation by TNF-alpha induced high levels of major histocompatibility complex class II and costimulatory molecules on DCs, but they remained weak producers of proinflammatory cytokines. One injection of such TNF/DCs pulsed with auto-antigenic peptide ameliorated the disease score of EAE. This could not be observed with immature DCs or DCs matured with lipopolysaccharide (LPS) plus anti-CD40. Three consecutive injections of peptide-pulsed TNF/DCs derived from wild-type led to the induction of peptide-specific predominantly interleukin (IL)-10-producing CD4(+) T cells and complete protection from EAE. Blocking of IL-10 in vivo could only partially restore the susceptibility to EAE, suggesting an important but not exclusive role of IL-10 for EAE prevention. Notably, the protection was peptide specific, as TNF/DCs pulsed with unrelated peptide could not prevent EAE. In conclusion, this study describes that stimulation by TNF-alpha results in incompletely matured DCs (semi-mature DCs) which induce peptide-specific IL-10-producing T cells in vivo and prevent EAE.

Show MeSH
Related in: MedlinePlus