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CXCL12 (SDF-1alpha) suppresses ongoing experimental autoimmune encephalomyelitis by selecting antigen-specific regulatory T cells.

Meiron M, Zohar Y, Anunu R, Wildbaum G, Karin N - J. Exp. Med. (2008)

Bottom Line: The beneficial effect included selection of antigen-specific T cells that were CD4(+)CD25(-)Foxp3(-)IL-10(high), which could adoptively transfer disease resistance, and suppression of Th17 selection.However, in vitro functional analysis of these cells suggested that, even though CXCL12-Ig-induced tolerance is IL-10 dependent, IL-10-independent mechanisms may also contribute to their regulatory function.Collectively, our results not only demonstrate, for the first time, that a chemokine functions as a regulatory mediator, but also suggest a novel way for treating multiple sclerosis and possibly other inflammatory autoimmune diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Bruce Rappaport Faculty of Medicine, Technion, Haifa 31096, Israel.

ABSTRACT
Experimental autoimmune encephalomyelitis (EAE) is a T cell-mediated autoimmune disease of the central nervous system induced by antigen-specific effector Th17 and Th1 cells. We show that a key chemokine, CXCL12 (stromal cell-derived factor 1alpha), redirects the polarization of effector Th1 cells into CD4(+)CD25(-)Foxp3(-)interleukin (IL) 10(high) antigen-specific regulatory T cells in a CXCR4-dependent manner, and by doing so acts as a regulatory mediator restraining the autoimmune inflammatory process. In an attempt to explore the therapeutic implication of these findings, we have generated a CXCL12-immunoglobulin (Ig) fusion protein that, when administered during ongoing EAE, rapidly suppresses the disease in wild-type but not IL-10-deficient mice. Anti-IL-10 neutralizing antibodies could reverse this suppression. The beneficial effect included selection of antigen-specific T cells that were CD4(+)CD25(-)Foxp3(-)IL-10(high), which could adoptively transfer disease resistance, and suppression of Th17 selection. However, in vitro functional analysis of these cells suggested that, even though CXCL12-Ig-induced tolerance is IL-10 dependent, IL-10-independent mechanisms may also contribute to their regulatory function. Collectively, our results not only demonstrate, for the first time, that a chemokine functions as a regulatory mediator, but also suggest a novel way for treating multiple sclerosis and possibly other inflammatory autoimmune diseases.

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CXCL12 directs the functional polarization of macrophages and T cells into high IL-10, low inflammatory mediator–producing cells. (A–C) CXCL12 was added at different concentrations to primary whole spleen culture taken from EAE mice and stimulated with their target MOGp35-55 antigen for 72 h (A), freshly isolated peritoneal macrophages stimulated with 0.5 μg/ml LPS for 48 h (B), or purified naive CD4+ T cells activated with anti-CD3/anti-CD28 for 48 h (C). Cytokine concentrations were measured in triplicates using a standard ELISA method. Results shown in this figure represent three independent experiments with similar results and are presented as means ± SE.
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fig2: CXCL12 directs the functional polarization of macrophages and T cells into high IL-10, low inflammatory mediator–producing cells. (A–C) CXCL12 was added at different concentrations to primary whole spleen culture taken from EAE mice and stimulated with their target MOGp35-55 antigen for 72 h (A), freshly isolated peritoneal macrophages stimulated with 0.5 μg/ml LPS for 48 h (B), or purified naive CD4+ T cells activated with anti-CD3/anti-CD28 for 48 h (C). Cytokine concentrations were measured in triplicates using a standard ELISA method. Results shown in this figure represent three independent experiments with similar results and are presented as means ± SE.

Mentions: How does CXCL12 suppress inflammation? We explored the possibility that this chemokine functions as a potential regulatory and antiinflammatory mediator. At first, rCXCL12 was added, in different concentrations, to primary spleen cells isolated from EAE mice that were cultured with their target antigen (myelin oligodendrocyte glycoprotein [MOG]p35-55). Fig. 2 A shows that supplementing these cultured cells with this chemokine induced an increase in IL-10 production (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), and at the same time a significant decrease in IL-12 and TNF-α production (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), all in a dose-dependent manner. Because the CXCR4 receptor is expressed on both CD4+ T cells and macrophages (1), we determined the direct effect of this chemokine on the production of various cytokines in each cell type separately. Initially, rCXCL12 was added in different concentrations to freshly isolated, LPS-activated peritoneal macrophages, and the effect on the production of various key cytokines was determined 48 h later. A dose-dependent increase in IL-10 production and, at the same time, a decrease in TNF-α and IL-12 were recorded (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12 for all comparisons; Fig. 2 B). To determine whether CXCL12 directly affects IL-10 production by CD4+ T cells, we isolated and purified CD4+ T cells from spleens of naive C57BL/6 mice (MACS beads) and subjected them to anti-CD3–induced activation in the presence of CXCL12. Fig. 2 C shows a significant, dose-dependent elevation in IL-10 production (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), accompanied by a significant reduction in TNF-α (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), and a dose-dependent increase in IL-2 production (Fig. 2 C). The increased production of IL-2 may suggest that the increased production of IL-10 results, in part, because of the differential proliferation of IL-10–producing cells, but it cannot explain the reduced levels of TNF-α produced by the cultured cells. These results may thus suggest a possible role for this chemokine as a regulatory mediator, and they motivated us to determine its potential use for the treatment of ongoing EAE.


CXCL12 (SDF-1alpha) suppresses ongoing experimental autoimmune encephalomyelitis by selecting antigen-specific regulatory T cells.

Meiron M, Zohar Y, Anunu R, Wildbaum G, Karin N - J. Exp. Med. (2008)

CXCL12 directs the functional polarization of macrophages and T cells into high IL-10, low inflammatory mediator–producing cells. (A–C) CXCL12 was added at different concentrations to primary whole spleen culture taken from EAE mice and stimulated with their target MOGp35-55 antigen for 72 h (A), freshly isolated peritoneal macrophages stimulated with 0.5 μg/ml LPS for 48 h (B), or purified naive CD4+ T cells activated with anti-CD3/anti-CD28 for 48 h (C). Cytokine concentrations were measured in triplicates using a standard ELISA method. Results shown in this figure represent three independent experiments with similar results and are presented as means ± SE.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2571938&req=5

fig2: CXCL12 directs the functional polarization of macrophages and T cells into high IL-10, low inflammatory mediator–producing cells. (A–C) CXCL12 was added at different concentrations to primary whole spleen culture taken from EAE mice and stimulated with their target MOGp35-55 antigen for 72 h (A), freshly isolated peritoneal macrophages stimulated with 0.5 μg/ml LPS for 48 h (B), or purified naive CD4+ T cells activated with anti-CD3/anti-CD28 for 48 h (C). Cytokine concentrations were measured in triplicates using a standard ELISA method. Results shown in this figure represent three independent experiments with similar results and are presented as means ± SE.
Mentions: How does CXCL12 suppress inflammation? We explored the possibility that this chemokine functions as a potential regulatory and antiinflammatory mediator. At first, rCXCL12 was added, in different concentrations, to primary spleen cells isolated from EAE mice that were cultured with their target antigen (myelin oligodendrocyte glycoprotein [MOG]p35-55). Fig. 2 A shows that supplementing these cultured cells with this chemokine induced an increase in IL-10 production (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), and at the same time a significant decrease in IL-12 and TNF-α production (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), all in a dose-dependent manner. Because the CXCR4 receptor is expressed on both CD4+ T cells and macrophages (1), we determined the direct effect of this chemokine on the production of various cytokines in each cell type separately. Initially, rCXCL12 was added in different concentrations to freshly isolated, LPS-activated peritoneal macrophages, and the effect on the production of various key cytokines was determined 48 h later. A dose-dependent increase in IL-10 production and, at the same time, a decrease in TNF-α and IL-12 were recorded (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12 for all comparisons; Fig. 2 B). To determine whether CXCL12 directly affects IL-10 production by CD4+ T cells, we isolated and purified CD4+ T cells from spleens of naive C57BL/6 mice (MACS beads) and subjected them to anti-CD3–induced activation in the presence of CXCL12. Fig. 2 C shows a significant, dose-dependent elevation in IL-10 production (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), accompanied by a significant reduction in TNF-α (P < 0.01 in cultures supplemented with 100 ng/ml CXCL12), and a dose-dependent increase in IL-2 production (Fig. 2 C). The increased production of IL-2 may suggest that the increased production of IL-10 results, in part, because of the differential proliferation of IL-10–producing cells, but it cannot explain the reduced levels of TNF-α produced by the cultured cells. These results may thus suggest a possible role for this chemokine as a regulatory mediator, and they motivated us to determine its potential use for the treatment of ongoing EAE.

Bottom Line: The beneficial effect included selection of antigen-specific T cells that were CD4(+)CD25(-)Foxp3(-)IL-10(high), which could adoptively transfer disease resistance, and suppression of Th17 selection.However, in vitro functional analysis of these cells suggested that, even though CXCL12-Ig-induced tolerance is IL-10 dependent, IL-10-independent mechanisms may also contribute to their regulatory function.Collectively, our results not only demonstrate, for the first time, that a chemokine functions as a regulatory mediator, but also suggest a novel way for treating multiple sclerosis and possibly other inflammatory autoimmune diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Bruce Rappaport Faculty of Medicine, Technion, Haifa 31096, Israel.

ABSTRACT
Experimental autoimmune encephalomyelitis (EAE) is a T cell-mediated autoimmune disease of the central nervous system induced by antigen-specific effector Th17 and Th1 cells. We show that a key chemokine, CXCL12 (stromal cell-derived factor 1alpha), redirects the polarization of effector Th1 cells into CD4(+)CD25(-)Foxp3(-)interleukin (IL) 10(high) antigen-specific regulatory T cells in a CXCR4-dependent manner, and by doing so acts as a regulatory mediator restraining the autoimmune inflammatory process. In an attempt to explore the therapeutic implication of these findings, we have generated a CXCL12-immunoglobulin (Ig) fusion protein that, when administered during ongoing EAE, rapidly suppresses the disease in wild-type but not IL-10-deficient mice. Anti-IL-10 neutralizing antibodies could reverse this suppression. The beneficial effect included selection of antigen-specific T cells that were CD4(+)CD25(-)Foxp3(-)IL-10(high), which could adoptively transfer disease resistance, and suppression of Th17 selection. However, in vitro functional analysis of these cells suggested that, even though CXCL12-Ig-induced tolerance is IL-10 dependent, IL-10-independent mechanisms may also contribute to their regulatory function. Collectively, our results not only demonstrate, for the first time, that a chemokine functions as a regulatory mediator, but also suggest a novel way for treating multiple sclerosis and possibly other inflammatory autoimmune diseases.

Show MeSH
Related in: MedlinePlus