Limits...
CD4(+)CD25(-)Foxp3(-) Th1 cells are the source of IL-10-mediated immune suppression in chronic cutaneous leishmaniasis.

Anderson CF, Oukka M, Kuchroo VJ, Sacks D - J. Exp. Med. (2007)

Bottom Line: Nonhealing forms of leishmaniasis in humans are commonly associated with elevated levels of the deactivating cytokine IL-10, and in the mouse, normally chronic infections can be cleared in the absence of IL-10.IL-10 was produced by innate cells, as well as CD4(+)CD25(+)Foxp3(+) and CD4(+)CD25(-)Foxp3(-) T cells in the chronic lesion.Surprisingly, Rag(-/-) mice reconstituted with naive CD4(+) T cells depleted of natural T regulatory cells developed more severe infections, associated with elevated levels of IL-10 and, especially, Th2 cytokines in the site.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.

ABSTRACT
Nonhealing forms of leishmaniasis in humans are commonly associated with elevated levels of the deactivating cytokine IL-10, and in the mouse, normally chronic infections can be cleared in the absence of IL-10. Using a Leishmania major strain that produces nonhealing dermal lesions in a T helper type 1 (Th1) cell-polarized setting, we have analyzed the cellular sources of IL-10 and their relative contribution to immune suppression. IL-10 was produced by innate cells, as well as CD4(+)CD25(+)Foxp3(+) and CD4(+)CD25(-)Foxp3(-) T cells in the chronic lesion. Nonetheless, only IL-10 production by antigen-specific CD4(+)CD25(-)Foxp3(-) T cells, the majority of which also produced IFN-gamma, was necessary for suppression of acquired immunity in Rag(-/-) reconstituted mice. Surprisingly, Rag(-/-) mice reconstituted with naive CD4(+) T cells depleted of natural T regulatory cells developed more severe infections, associated with elevated levels of IL-10 and, especially, Th2 cytokines in the site. The data demonstrate that IL-10-producing Th1 cells, activated early in a strong inflammatory setting as a mechanism of feedback control, are the principal mediators of T cell-derived IL-10-dependent immune suppression in a chronic intracellular infection.

Show MeSH

Related in: MedlinePlus

CD4+ T cells producing IL-10 in response to infection with L. major NIH/Sd comprise multiple subsets. (A) Naive ear cells or cells from the chronic ear lesion were stimulated in vitro with PMA and ionomycin for 4 h and analyzed for intracellular cytokine production. (B) A kinetic analysis of IL-10 production and Foxp3 expression was performed on in vitro–restimulated lesional cells at the indicated time points. The data shown are TCRβ+CD4+ cells from six pooled ears from one experiment and are representative of at least three independent experiments. Quadrant values are the percentage of total gated CD4+ T cells. (C) 10 wk after infection, TCRβ+CD4+ ear lesion cells were purified by cell sorting into three subsets according to cell surface expression of CD25 and CD103. Real-time PCR analysis was performed to quantify gene expression of Foxp3, IFN-γ, and IL-10. Data are expressed as the fold change relative to naive CD4+CD25− lymph node cells. Two independent experiments were performed with similar results.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2118728&req=5

fig2: CD4+ T cells producing IL-10 in response to infection with L. major NIH/Sd comprise multiple subsets. (A) Naive ear cells or cells from the chronic ear lesion were stimulated in vitro with PMA and ionomycin for 4 h and analyzed for intracellular cytokine production. (B) A kinetic analysis of IL-10 production and Foxp3 expression was performed on in vitro–restimulated lesional cells at the indicated time points. The data shown are TCRβ+CD4+ cells from six pooled ears from one experiment and are representative of at least three independent experiments. Quadrant values are the percentage of total gated CD4+ T cells. (C) 10 wk after infection, TCRβ+CD4+ ear lesion cells were purified by cell sorting into three subsets according to cell surface expression of CD25 and CD103. Real-time PCR analysis was performed to quantify gene expression of Foxp3, IFN-γ, and IL-10. Data are expressed as the fold change relative to naive CD4+CD25− lymph node cells. Two independent experiments were performed with similar results.

Mentions: PMA/ionomycin stimulation of naive dermal CD4+ T cells indicated that all of the IL-10 production was confined to the resident CD25+ cells (Fig. 2 A). In contrast, in the cells recovered from the chronic lesion (10 wk), IL-10 was produced by approximately equivalent numbers of CD25− and CD25+ cells, whereas the vast majority of the IFN-γ–producing cells were CD25−. Foxp3 staining, which provides a more stringent criterion for identification of natural T reg cells, indicated that at 3 wk after infection, ∼15% of the lesional CD4+ cells expressed Foxp3, and this frequency was more or less maintained throughout the chronic phase of disease (Fig. 2 B). Of note, a substantial number of the Foxp3+ cells were CD25−. IL-10 production by the CD25− cells was not confined to the Foxp3+ population, however, because CD4+Foxp3+ and CD4+Foxp3− cells were each a source of IL-10 in the lesion. Kinetic analysis of IL-10 production by CD4+Foxp3− and CD4+Foxp3+ cells showed that both populations were present during the early phase of the effector response (3 wk after infection), suggesting that these cells are not a consequence of chronic antigen stimulation in the inflammatory site but arise early and may be causally related to the evolution of the noncure phenotype.


CD4(+)CD25(-)Foxp3(-) Th1 cells are the source of IL-10-mediated immune suppression in chronic cutaneous leishmaniasis.

Anderson CF, Oukka M, Kuchroo VJ, Sacks D - J. Exp. Med. (2007)

CD4+ T cells producing IL-10 in response to infection with L. major NIH/Sd comprise multiple subsets. (A) Naive ear cells or cells from the chronic ear lesion were stimulated in vitro with PMA and ionomycin for 4 h and analyzed for intracellular cytokine production. (B) A kinetic analysis of IL-10 production and Foxp3 expression was performed on in vitro–restimulated lesional cells at the indicated time points. The data shown are TCRβ+CD4+ cells from six pooled ears from one experiment and are representative of at least three independent experiments. Quadrant values are the percentage of total gated CD4+ T cells. (C) 10 wk after infection, TCRβ+CD4+ ear lesion cells were purified by cell sorting into three subsets according to cell surface expression of CD25 and CD103. Real-time PCR analysis was performed to quantify gene expression of Foxp3, IFN-γ, and IL-10. Data are expressed as the fold change relative to naive CD4+CD25− lymph node cells. Two independent experiments were performed with similar results.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2118728&req=5

fig2: CD4+ T cells producing IL-10 in response to infection with L. major NIH/Sd comprise multiple subsets. (A) Naive ear cells or cells from the chronic ear lesion were stimulated in vitro with PMA and ionomycin for 4 h and analyzed for intracellular cytokine production. (B) A kinetic analysis of IL-10 production and Foxp3 expression was performed on in vitro–restimulated lesional cells at the indicated time points. The data shown are TCRβ+CD4+ cells from six pooled ears from one experiment and are representative of at least three independent experiments. Quadrant values are the percentage of total gated CD4+ T cells. (C) 10 wk after infection, TCRβ+CD4+ ear lesion cells were purified by cell sorting into three subsets according to cell surface expression of CD25 and CD103. Real-time PCR analysis was performed to quantify gene expression of Foxp3, IFN-γ, and IL-10. Data are expressed as the fold change relative to naive CD4+CD25− lymph node cells. Two independent experiments were performed with similar results.
Mentions: PMA/ionomycin stimulation of naive dermal CD4+ T cells indicated that all of the IL-10 production was confined to the resident CD25+ cells (Fig. 2 A). In contrast, in the cells recovered from the chronic lesion (10 wk), IL-10 was produced by approximately equivalent numbers of CD25− and CD25+ cells, whereas the vast majority of the IFN-γ–producing cells were CD25−. Foxp3 staining, which provides a more stringent criterion for identification of natural T reg cells, indicated that at 3 wk after infection, ∼15% of the lesional CD4+ cells expressed Foxp3, and this frequency was more or less maintained throughout the chronic phase of disease (Fig. 2 B). Of note, a substantial number of the Foxp3+ cells were CD25−. IL-10 production by the CD25− cells was not confined to the Foxp3+ population, however, because CD4+Foxp3+ and CD4+Foxp3− cells were each a source of IL-10 in the lesion. Kinetic analysis of IL-10 production by CD4+Foxp3− and CD4+Foxp3+ cells showed that both populations were present during the early phase of the effector response (3 wk after infection), suggesting that these cells are not a consequence of chronic antigen stimulation in the inflammatory site but arise early and may be causally related to the evolution of the noncure phenotype.

Bottom Line: Nonhealing forms of leishmaniasis in humans are commonly associated with elevated levels of the deactivating cytokine IL-10, and in the mouse, normally chronic infections can be cleared in the absence of IL-10.IL-10 was produced by innate cells, as well as CD4(+)CD25(+)Foxp3(+) and CD4(+)CD25(-)Foxp3(-) T cells in the chronic lesion.Surprisingly, Rag(-/-) mice reconstituted with naive CD4(+) T cells depleted of natural T regulatory cells developed more severe infections, associated with elevated levels of IL-10 and, especially, Th2 cytokines in the site.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.

ABSTRACT
Nonhealing forms of leishmaniasis in humans are commonly associated with elevated levels of the deactivating cytokine IL-10, and in the mouse, normally chronic infections can be cleared in the absence of IL-10. Using a Leishmania major strain that produces nonhealing dermal lesions in a T helper type 1 (Th1) cell-polarized setting, we have analyzed the cellular sources of IL-10 and their relative contribution to immune suppression. IL-10 was produced by innate cells, as well as CD4(+)CD25(+)Foxp3(+) and CD4(+)CD25(-)Foxp3(-) T cells in the chronic lesion. Nonetheless, only IL-10 production by antigen-specific CD4(+)CD25(-)Foxp3(-) T cells, the majority of which also produced IFN-gamma, was necessary for suppression of acquired immunity in Rag(-/-) reconstituted mice. Surprisingly, Rag(-/-) mice reconstituted with naive CD4(+) T cells depleted of natural T regulatory cells developed more severe infections, associated with elevated levels of IL-10 and, especially, Th2 cytokines in the site. The data demonstrate that IL-10-producing Th1 cells, activated early in a strong inflammatory setting as a mechanism of feedback control, are the principal mediators of T cell-derived IL-10-dependent immune suppression in a chronic intracellular infection.

Show MeSH
Related in: MedlinePlus