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B7-H1-deficiency enhances the potential of tolerogenic dendritic cells by activating CD1d-restricted type II NKT cells.

Brandl C, Ortler S, Herrmann T, Cardell S, Lutz MB, Wiendl H - PLoS ONE (2010)

Bottom Line: Injections of B7-H1-deficient DC showed increased EAE protection as compared to wild type (WT)-DC.Injections of B7-H1(-/-) TNF-DC induced higher release of peptide-specific IL-10 and IL-13 after restimulation in vitro together with elevated serum cytokines IL-4 and IL-13 produced by NKT cells, and reduced IL-17 and IFN-gamma production in the CNS.In the absence of B7-H1 on these DC their tolerogenic potential to stimulate tolerogenic CD4(+) and NKT cell responses is enhanced.

View Article: PubMed Central - PubMed

Affiliation: Institute of Virology and Immunobiology, University of Wuerzburg, Wuerzburg, Germany.

ABSTRACT

Background: Dendritic cells (DC) can act tolerogenic at a semi-mature stage by induction of protective CD4(+) T cell and NKT cell responses.

Methodology/principal findings: Here we studied the role of the co-inhibitory molecule B7-H1 (PD-L1, CD274) on semi-mature DC that were generated from bone marrow (BM) cells of B7-H1(-/-) mice and applied to the model of Experimental Autoimmune Encephalomyelitis (EAE). Injections of B7-H1-deficient DC showed increased EAE protection as compared to wild type (WT)-DC. Injections of B7-H1(-/-) TNF-DC induced higher release of peptide-specific IL-10 and IL-13 after restimulation in vitro together with elevated serum cytokines IL-4 and IL-13 produced by NKT cells, and reduced IL-17 and IFN-gamma production in the CNS. Experiments in CD1d(-/-) and Jalpha281(-/-) mice as well as with type I and II NKT cell lines indicated that only type II NKT cells but not type I NKT cells (invariant NKT cells) could be stimulated by an endogenous CD1d-ligand on DC and were responsible for the increased serum cytokine production in the absence of B7-H1.

Conclusions/significance: Together, our data indicate that BM-DC express an endogenous CD1d ligand and B7-H1 to ihibit type II but not type I NKT cells. In the absence of B7-H1 on these DC their tolerogenic potential to stimulate tolerogenic CD4(+) and NKT cell responses is enhanced.

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Strong increase of protective serum cytokines after injection of B7-H1āˆ’/āˆ’ DC injection is mainly produced by type II NKT cells.A) Splenocytes from WT and JĪ±281āˆ’/āˆ’ mice were stained for CD3 and NK1.1 and investigated by flow cytometry. Double positive cells (NKT cells) were analyzed for PD-1 expression. B) MOG35ā€“55 loaded and TNF-matured WT or B7-H1āˆ’/āˆ’ DC (2,5Ɨ106) were injected i.v. into WT, CD1dāˆ’/āˆ’ or JĪ±281āˆ’/āˆ’ mice three times (on day āˆ’4, āˆ’2 and 0). Similarly, control mice were injected with PBS. Blood was collected 2 hours after the last DC injection and serum cytokines were measured by ELISA for IFN-Ī³, IL-17, IL-4 and IL-13. The results represent the average of 3 mice per group and are representative for 4 independent experiments for the WT mice and for 2 independent experiments for CD1dāˆ’/āˆ’ and JĪ±281āˆ’/āˆ’ mice. * p<0.05, ** p<0.01, *** p<0.001, n.s. not significant.
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pone-0010800-g005: Strong increase of protective serum cytokines after injection of B7-H1āˆ’/āˆ’ DC injection is mainly produced by type II NKT cells.A) Splenocytes from WT and JĪ±281āˆ’/āˆ’ mice were stained for CD3 and NK1.1 and investigated by flow cytometry. Double positive cells (NKT cells) were analyzed for PD-1 expression. B) MOG35ā€“55 loaded and TNF-matured WT or B7-H1āˆ’/āˆ’ DC (2,5Ɨ106) were injected i.v. into WT, CD1dāˆ’/āˆ’ or JĪ±281āˆ’/āˆ’ mice three times (on day āˆ’4, āˆ’2 and 0). Similarly, control mice were injected with PBS. Blood was collected 2 hours after the last DC injection and serum cytokines were measured by ELISA for IFN-Ī³, IL-17, IL-4 and IL-13. The results represent the average of 3 mice per group and are representative for 4 independent experiments for the WT mice and for 2 independent experiments for CD1dāˆ’/āˆ’ and JĪ±281āˆ’/āˆ’ mice. * p<0.05, ** p<0.01, *** p<0.001, n.s. not significant.

Mentions: We showed previously that TNF-DC induce a rapid production of type 2 cytokines detectable in mouse sera early after the third injection of TNF-DC and their contribution to EAE prevention by constraining Th1 and Th17 effector cell development [2]. As a prerequisite we tested the PD-1 expression on the cell surface by type I and II NKT cells of WT mice and the type II NKT cells of JĪ±281āˆ’/āˆ’ mice and could not find major differences (Figure 5A). To determine the relevance of TNF-DC B7-H1 expression for protective cytokine production, sera from WT or B7-H1āˆ’/āˆ’ TNF-DC-injected mice were collected and analyzed for cytokine profile by ELISA. While IL-17 concentration was generally low and remained unaffected by TNF-DC treatment (data not shown), TNF-DC-injected mice displayed significantly elevated levels of IFN-Ī³ in their sera (Figure 5B). This effect was even reinforced in the absence of B7-H1 on TNF-DC. Simultaneously, IL-4 and IL-13 concentrations were markedly increased in WT TNF-DC-injected mice compared to control mice but significantly amplified using B7-H1-deficient TNF-DC. These findings are in line with the reduced disease course and support the notion of an increased induction of IL-4 and IL-13 producing cells in the absence of inhibitory B7-H1 on DC.


B7-H1-deficiency enhances the potential of tolerogenic dendritic cells by activating CD1d-restricted type II NKT cells.

Brandl C, Ortler S, Herrmann T, Cardell S, Lutz MB, Wiendl H - PLoS ONE (2010)

Strong increase of protective serum cytokines after injection of B7-H1āˆ’/āˆ’ DC injection is mainly produced by type II NKT cells.A) Splenocytes from WT and JĪ±281āˆ’/āˆ’ mice were stained for CD3 and NK1.1 and investigated by flow cytometry. Double positive cells (NKT cells) were analyzed for PD-1 expression. B) MOG35ā€“55 loaded and TNF-matured WT or B7-H1āˆ’/āˆ’ DC (2,5Ɨ106) were injected i.v. into WT, CD1dāˆ’/āˆ’ or JĪ±281āˆ’/āˆ’ mice three times (on day āˆ’4, āˆ’2 and 0). Similarly, control mice were injected with PBS. Blood was collected 2 hours after the last DC injection and serum cytokines were measured by ELISA for IFN-Ī³, IL-17, IL-4 and IL-13. The results represent the average of 3 mice per group and are representative for 4 independent experiments for the WT mice and for 2 independent experiments for CD1dāˆ’/āˆ’ and JĪ±281āˆ’/āˆ’ mice. * p<0.05, ** p<0.01, *** p<0.001, n.s. not significant.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2875405&req=5

pone-0010800-g005: Strong increase of protective serum cytokines after injection of B7-H1āˆ’/āˆ’ DC injection is mainly produced by type II NKT cells.A) Splenocytes from WT and JĪ±281āˆ’/āˆ’ mice were stained for CD3 and NK1.1 and investigated by flow cytometry. Double positive cells (NKT cells) were analyzed for PD-1 expression. B) MOG35ā€“55 loaded and TNF-matured WT or B7-H1āˆ’/āˆ’ DC (2,5Ɨ106) were injected i.v. into WT, CD1dāˆ’/āˆ’ or JĪ±281āˆ’/āˆ’ mice three times (on day āˆ’4, āˆ’2 and 0). Similarly, control mice were injected with PBS. Blood was collected 2 hours after the last DC injection and serum cytokines were measured by ELISA for IFN-Ī³, IL-17, IL-4 and IL-13. The results represent the average of 3 mice per group and are representative for 4 independent experiments for the WT mice and for 2 independent experiments for CD1dāˆ’/āˆ’ and JĪ±281āˆ’/āˆ’ mice. * p<0.05, ** p<0.01, *** p<0.001, n.s. not significant.
Mentions: We showed previously that TNF-DC induce a rapid production of type 2 cytokines detectable in mouse sera early after the third injection of TNF-DC and their contribution to EAE prevention by constraining Th1 and Th17 effector cell development [2]. As a prerequisite we tested the PD-1 expression on the cell surface by type I and II NKT cells of WT mice and the type II NKT cells of JĪ±281āˆ’/āˆ’ mice and could not find major differences (Figure 5A). To determine the relevance of TNF-DC B7-H1 expression for protective cytokine production, sera from WT or B7-H1āˆ’/āˆ’ TNF-DC-injected mice were collected and analyzed for cytokine profile by ELISA. While IL-17 concentration was generally low and remained unaffected by TNF-DC treatment (data not shown), TNF-DC-injected mice displayed significantly elevated levels of IFN-Ī³ in their sera (Figure 5B). This effect was even reinforced in the absence of B7-H1 on TNF-DC. Simultaneously, IL-4 and IL-13 concentrations were markedly increased in WT TNF-DC-injected mice compared to control mice but significantly amplified using B7-H1-deficient TNF-DC. These findings are in line with the reduced disease course and support the notion of an increased induction of IL-4 and IL-13 producing cells in the absence of inhibitory B7-H1 on DC.

Bottom Line: Injections of B7-H1-deficient DC showed increased EAE protection as compared to wild type (WT)-DC.Injections of B7-H1(-/-) TNF-DC induced higher release of peptide-specific IL-10 and IL-13 after restimulation in vitro together with elevated serum cytokines IL-4 and IL-13 produced by NKT cells, and reduced IL-17 and IFN-gamma production in the CNS.In the absence of B7-H1 on these DC their tolerogenic potential to stimulate tolerogenic CD4(+) and NKT cell responses is enhanced.

View Article: PubMed Central - PubMed

Affiliation: Institute of Virology and Immunobiology, University of Wuerzburg, Wuerzburg, Germany.

ABSTRACT

Background: Dendritic cells (DC) can act tolerogenic at a semi-mature stage by induction of protective CD4(+) T cell and NKT cell responses.

Methodology/principal findings: Here we studied the role of the co-inhibitory molecule B7-H1 (PD-L1, CD274) on semi-mature DC that were generated from bone marrow (BM) cells of B7-H1(-/-) mice and applied to the model of Experimental Autoimmune Encephalomyelitis (EAE). Injections of B7-H1-deficient DC showed increased EAE protection as compared to wild type (WT)-DC. Injections of B7-H1(-/-) TNF-DC induced higher release of peptide-specific IL-10 and IL-13 after restimulation in vitro together with elevated serum cytokines IL-4 and IL-13 produced by NKT cells, and reduced IL-17 and IFN-gamma production in the CNS. Experiments in CD1d(-/-) and Jalpha281(-/-) mice as well as with type I and II NKT cell lines indicated that only type II NKT cells but not type I NKT cells (invariant NKT cells) could be stimulated by an endogenous CD1d-ligand on DC and were responsible for the increased serum cytokine production in the absence of B7-H1.

Conclusions/significance: Together, our data indicate that BM-DC express an endogenous CD1d ligand and B7-H1 to ihibit type II but not type I NKT cells. In the absence of B7-H1 on these DC their tolerogenic potential to stimulate tolerogenic CD4(+) and NKT cell responses is enhanced.

Show MeSH
Related in: MedlinePlus