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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 induce peptide-specific CD4+ T cells producing IL-10 which is partially involved in EAE protection. (a) TNF/DCs induce IL-10–producing cells from restimulated spleen cells. C57BL/6 mice received three intravenous injections of MOG-pulsed TNF/DCs or LPS/CD40/DCs at days −7, −5, and −3. Spleen cells from these mice were restimulated at day 0 with 10 μM MOG peptide or 10 μM unrelated OVA peptide. Cell supernatants were taken after 24, 48, 72, and 96 h and tested for their cytokine content by ELISA. (b) The IL-10 production from spleen cells induced by DC injections is derived from CD4+CD3+ T cells, but not CD11c+ DCs. C57BL/6 mice received three injections of MOG pulsed TNF/DC at days −7, −5, and −3. Spleen cells from these mice were sorted for CD4+ and CD11c+ cells at day 0 and then stimulated for 24 h with PMA plus Ionomycin before supernatants were tested by ELISA for their IL-10 content. From the CD4+ enriched cells 97% coexpressed CD3, and the contamination CD11c+ cells within the CD4+ cells was 0.8–1.4%. (c) The protection from EAE partially depends on IL-10. C57BL/6 mice (3–4 per group) were injected three times with MOG/TNF/DCs at days −7, −5, and −3 (3×) and EAE was induced at day 0. Mice were injected intraperitoneally with anti–IL-10R mAb or isotype mAb at the same days as TNF/DCs and additionally at day −1 and day 1. Control mice remained without pretreatment by DCs and were not injected with mAb. The data for (a) is representative of three, and the data for b and c are each representative of two independent experiments with similar results.
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fig3: Repetitive injections of TNF/DCs induce peptide-specific CD4+ T cells producing IL-10 which is partially involved in EAE protection. (a) TNF/DCs induce IL-10–producing cells from restimulated spleen cells. C57BL/6 mice received three intravenous injections of MOG-pulsed TNF/DCs or LPS/CD40/DCs at days −7, −5, and −3. Spleen cells from these mice were restimulated at day 0 with 10 μM MOG peptide or 10 μM unrelated OVA peptide. Cell supernatants were taken after 24, 48, 72, and 96 h and tested for their cytokine content by ELISA. (b) The IL-10 production from spleen cells induced by DC injections is derived from CD4+CD3+ T cells, but not CD11c+ DCs. C57BL/6 mice received three injections of MOG pulsed TNF/DC at days −7, −5, and −3. Spleen cells from these mice were sorted for CD4+ and CD11c+ cells at day 0 and then stimulated for 24 h with PMA plus Ionomycin before supernatants were tested by ELISA for their IL-10 content. From the CD4+ enriched cells 97% coexpressed CD3, and the contamination CD11c+ cells within the CD4+ cells was 0.8–1.4%. (c) The protection from EAE partially depends on IL-10. C57BL/6 mice (3–4 per group) were injected three times with MOG/TNF/DCs at days −7, −5, and −3 (3×) and EAE was induced at day 0. Mice were injected intraperitoneally with anti–IL-10R mAb or isotype mAb at the same days as TNF/DCs and additionally at day −1 and day 1. Control mice remained without pretreatment by DCs and were not injected with mAb. The data for (a) is representative of three, and the data for b and c are each representative of two independent experiments with similar results.

Mentions: To assess the mechanism of complete EAE suppression in mice that received three injections with MOG/TNF/DCs or MOG/LPS/CD40/DCs their spleens were removed at day 0 and restimulated with MOG peptide or irrelevant OVA peptide, and the culture supernatants were analyzed after 24, 48, 72, and 96 h for their cytokine content. The predominant cytokine that could be detected after three MOG/TNF/DCs injections was IL-10, whereas only little IFN-γ and no IL-2 or IL-4 were present (Fig. 3 a). Preliminary data indicate that also TGF-β1 is absent (not shown). In contrast, three injections of MOG/LPS/CD40/DCs induced predominantly IFN-γ but no IL-2, IL-4, or IL-10, indicative for the induction of a Th1 response, which could explain why such DCs are not protective in the Th1-mediated EAE.


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 induce peptide-specific CD4+ T cells producing IL-10 which is partially involved in EAE protection. (a) TNF/DCs induce IL-10–producing cells from restimulated spleen cells. C57BL/6 mice received three intravenous injections of MOG-pulsed TNF/DCs or LPS/CD40/DCs at days −7, −5, and −3. Spleen cells from these mice were restimulated at day 0 with 10 μM MOG peptide or 10 μM unrelated OVA peptide. Cell supernatants were taken after 24, 48, 72, and 96 h and tested for their cytokine content by ELISA. (b) The IL-10 production from spleen cells induced by DC injections is derived from CD4+CD3+ T cells, but not CD11c+ DCs. C57BL/6 mice received three injections of MOG pulsed TNF/DC at days −7, −5, and −3. Spleen cells from these mice were sorted for CD4+ and CD11c+ cells at day 0 and then stimulated for 24 h with PMA plus Ionomycin before supernatants were tested by ELISA for their IL-10 content. From the CD4+ enriched cells 97% coexpressed CD3, and the contamination CD11c+ cells within the CD4+ cells was 0.8–1.4%. (c) The protection from EAE partially depends on IL-10. C57BL/6 mice (3–4 per group) were injected three times with MOG/TNF/DCs at days −7, −5, and −3 (3×) and EAE was induced at day 0. Mice were injected intraperitoneally with anti–IL-10R mAb or isotype mAb at the same days as TNF/DCs and additionally at day −1 and day 1. Control mice remained without pretreatment by DCs and were not injected with mAb. The data for (a) is representative of three, and the data for b and c are each representative of two independent experiments with similar results.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196016&req=5

fig3: Repetitive injections of TNF/DCs induce peptide-specific CD4+ T cells producing IL-10 which is partially involved in EAE protection. (a) TNF/DCs induce IL-10–producing cells from restimulated spleen cells. C57BL/6 mice received three intravenous injections of MOG-pulsed TNF/DCs or LPS/CD40/DCs at days −7, −5, and −3. Spleen cells from these mice were restimulated at day 0 with 10 μM MOG peptide or 10 μM unrelated OVA peptide. Cell supernatants were taken after 24, 48, 72, and 96 h and tested for their cytokine content by ELISA. (b) The IL-10 production from spleen cells induced by DC injections is derived from CD4+CD3+ T cells, but not CD11c+ DCs. C57BL/6 mice received three injections of MOG pulsed TNF/DC at days −7, −5, and −3. Spleen cells from these mice were sorted for CD4+ and CD11c+ cells at day 0 and then stimulated for 24 h with PMA plus Ionomycin before supernatants were tested by ELISA for their IL-10 content. From the CD4+ enriched cells 97% coexpressed CD3, and the contamination CD11c+ cells within the CD4+ cells was 0.8–1.4%. (c) The protection from EAE partially depends on IL-10. C57BL/6 mice (3–4 per group) were injected three times with MOG/TNF/DCs at days −7, −5, and −3 (3×) and EAE was induced at day 0. Mice were injected intraperitoneally with anti–IL-10R mAb or isotype mAb at the same days as TNF/DCs and additionally at day −1 and day 1. Control mice remained without pretreatment by DCs and were not injected with mAb. The data for (a) is representative of three, and the data for b and c are each representative of two independent experiments with similar results.
Mentions: To assess the mechanism of complete EAE suppression in mice that received three injections with MOG/TNF/DCs or MOG/LPS/CD40/DCs their spleens were removed at day 0 and restimulated with MOG peptide or irrelevant OVA peptide, and the culture supernatants were analyzed after 24, 48, 72, and 96 h for their cytokine content. The predominant cytokine that could be detected after three MOG/TNF/DCs injections was IL-10, whereas only little IFN-γ and no IL-2 or IL-4 were present (Fig. 3 a). Preliminary data indicate that also TGF-β1 is absent (not shown). In contrast, three injections of MOG/LPS/CD40/DCs induced predominantly IFN-γ but no IL-2, IL-4, or IL-10, indicative for the induction of a Th1 response, which could explain why such DCs are not protective in the Th1-mediated EAE.

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