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Antigen-Specific Th17 Cells Are Primed by Distinct and Complementary Dendritic Cell Subsets in Oropharyngeal Candidiasis.

Trautwein-Weidner K, Gladiator A, Kirchner FR, Becattini S, Rülicke T, Sallusto F, LeibundGut-Landmann S - PLoS Pathog. (2015)

Bottom Line: The events regulating T cell activation and differentiation toward effector fates in response to fungal invasion in different tissues are poorly understood.This highlights the functional compartmentalization of specific DC subsets in different tissues.These data provide important new insights to our understanding of tissue-specific antifungal immunity.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microbiology, ETH Zürich, Zürich, Switzerland.

ABSTRACT
Candida spp. can cause severe and chronic mucocutaneous and systemic infections in immunocompromised individuals. Protection from mucocutaneous candidiasis depends on T helper cells, in particular those secreting IL-17. The events regulating T cell activation and differentiation toward effector fates in response to fungal invasion in different tissues are poorly understood. Here we generated a Candida-specific TCR transgenic mouse reactive to a novel endogenous antigen that is conserved in multiple distant species of Candida, including the clinically highly relevant C. albicans and C. glabrata. Using TCR transgenic T cells in combination with an experimental model of oropharyngeal candidiasis (OPC) we investigated antigen presentation and Th17 priming by different subsets of dendritic cells (DCs) present in the infected oral mucosa. Candida-derived endogenous antigen accesses the draining lymph nodes and is directly presented by migratory DCs. Tissue-resident Flt3L-dependent DCs and CCR2-dependent monocyte-derived DCs collaborate in antigen presentation and T cell priming during OPC. In contrast, Langerhans cells, which are also present in the oral mucosa and have been shown to prime Th17 cells in the skin, are not required for induction of the Candida-specific T cell response upon oral challenge. This highlights the functional compartmentalization of specific DC subsets in different tissues. These data provide important new insights to our understanding of tissue-specific antifungal immunity.

No MeSH data available.


Related in: MedlinePlus

Flt3L-dependent migratory DCs and monocyte-derived DCs both present C. albicans derived antigen.(A) Cervical lymph node cells of B6 or Flt3l-/- mice were analyzed on day 2 post-infection. Representative FACS plots from individual mice and quantification of MHC IIhigh CD11c+ (population I), MHC II+ CD11chigh (population II) and MHC II+ CD11c+ cells (population III) from one of two independent experiments are shown. (B, C) Cervical lymph node cells were isolated from naïve B6 mice and from infected B6 and Flt3l-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were analyzed for CD69 expression after 1 day (B) and for proliferation after 4 days, respectively (C). Representative plots are shown on the left, summary of data from individual mice with mean + SD are shown on the right. Each symbol represents one mouse. Data are representative of 2 independent experiments. (D) As in B, but MHC IIhigh CD11c+ migratory DCs were isolated by FACS-sorting from the cervical lymph nodes of infected B6 and Flt3l-/- mice. (E—F) CCR2+ CD11b+ cells in the cervical lymph nodes of naïve and infected B6 mice on day 2 post-infection were analyzed for the expression of MHC II and CD11c (D). Three distinct subsets of CCR2+ CD11b+ cells (MHC IIlo/int Ly6C+ (Q1), MHC IIint Ly6C- (Q2) and MHC IIhigh Ly6C- (Q3)) from infected mice were further analyzed for the expression of CCR2 and CD11c (E). Representative FACS plots are shown. (G) Cervical lymph nodes were isolated from infected B6 mice on day 2 post-infection. CCR2hi, CCR2int and CCR2lo subsets within the MHC IIhigh CD11c+ population I were FACS-sorted and co-cultured with CD4+ Hector T cells for 1 day. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were then analyzed for CD69 expression. Representative FACS plots are shown, Data are representative of 2 independent experiments.
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ppat.1005164.g006: Flt3L-dependent migratory DCs and monocyte-derived DCs both present C. albicans derived antigen.(A) Cervical lymph node cells of B6 or Flt3l-/- mice were analyzed on day 2 post-infection. Representative FACS plots from individual mice and quantification of MHC IIhigh CD11c+ (population I), MHC II+ CD11chigh (population II) and MHC II+ CD11c+ cells (population III) from one of two independent experiments are shown. (B, C) Cervical lymph node cells were isolated from naïve B6 mice and from infected B6 and Flt3l-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were analyzed for CD69 expression after 1 day (B) and for proliferation after 4 days, respectively (C). Representative plots are shown on the left, summary of data from individual mice with mean + SD are shown on the right. Each symbol represents one mouse. Data are representative of 2 independent experiments. (D) As in B, but MHC IIhigh CD11c+ migratory DCs were isolated by FACS-sorting from the cervical lymph nodes of infected B6 and Flt3l-/- mice. (E—F) CCR2+ CD11b+ cells in the cervical lymph nodes of naïve and infected B6 mice on day 2 post-infection were analyzed for the expression of MHC II and CD11c (D). Three distinct subsets of CCR2+ CD11b+ cells (MHC IIlo/int Ly6C+ (Q1), MHC IIint Ly6C- (Q2) and MHC IIhigh Ly6C- (Q3)) from infected mice were further analyzed for the expression of CCR2 and CD11c (E). Representative FACS plots are shown. (G) Cervical lymph nodes were isolated from infected B6 mice on day 2 post-infection. CCR2hi, CCR2int and CCR2lo subsets within the MHC IIhigh CD11c+ population I were FACS-sorted and co-cultured with CD4+ Hector T cells for 1 day. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were then analyzed for CD69 expression. Representative FACS plots are shown, Data are representative of 2 independent experiments.

Mentions: The large Langerin−CD103– subset of migratory DCs in the cervical lymph node appeared to be phenotypically homogeneous for all markers analyzed (Fig 4B). However, the subset may still comprise phylogenetically and functionally distinct cell types, including Flt3-dependent conventional DCs and monocyte-derived DCs that are Flt3-independent but dependent on Csf1R signaling for differentiation from inflammatory monocyte [13]. Consistent with this notion, we found that in Flt3l-/- mice, migratory MHC IIhi CD11c+ DCs, which include the large population of Langerin−CD103– cells, were strongly reduced in Flt3l-/- mice compared to B6 mice (Fig 6A). Furthermore, the CD11b+ cervical lymph node cells isolated from infected Flt3l-/- mice were strongly impaired in their ability to induce CD69 upregulation and proliferation of Hector T cells in vitro (Fig 6B and 6C). This became also clear when the APC fraction was purified by FACS sorting: antigen presentation by Flt3L-dependent migratory DCs strongly promoted the activation of CD4+ Hector T cells in vitro, it was however not essential for the response (Fig 6D), indicating that Flt3L-independent migratory DCs are also involved. Thus, Flt3L-dependent migratory DCs appear to be an important source of antigen in cervical lymph nodes for T cell activation in response to C. albicans oral infection.


Antigen-Specific Th17 Cells Are Primed by Distinct and Complementary Dendritic Cell Subsets in Oropharyngeal Candidiasis.

Trautwein-Weidner K, Gladiator A, Kirchner FR, Becattini S, Rülicke T, Sallusto F, LeibundGut-Landmann S - PLoS Pathog. (2015)

Flt3L-dependent migratory DCs and monocyte-derived DCs both present C. albicans derived antigen.(A) Cervical lymph node cells of B6 or Flt3l-/- mice were analyzed on day 2 post-infection. Representative FACS plots from individual mice and quantification of MHC IIhigh CD11c+ (population I), MHC II+ CD11chigh (population II) and MHC II+ CD11c+ cells (population III) from one of two independent experiments are shown. (B, C) Cervical lymph node cells were isolated from naïve B6 mice and from infected B6 and Flt3l-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were analyzed for CD69 expression after 1 day (B) and for proliferation after 4 days, respectively (C). Representative plots are shown on the left, summary of data from individual mice with mean + SD are shown on the right. Each symbol represents one mouse. Data are representative of 2 independent experiments. (D) As in B, but MHC IIhigh CD11c+ migratory DCs were isolated by FACS-sorting from the cervical lymph nodes of infected B6 and Flt3l-/- mice. (E—F) CCR2+ CD11b+ cells in the cervical lymph nodes of naïve and infected B6 mice on day 2 post-infection were analyzed for the expression of MHC II and CD11c (D). Three distinct subsets of CCR2+ CD11b+ cells (MHC IIlo/int Ly6C+ (Q1), MHC IIint Ly6C- (Q2) and MHC IIhigh Ly6C- (Q3)) from infected mice were further analyzed for the expression of CCR2 and CD11c (E). Representative FACS plots are shown. (G) Cervical lymph nodes were isolated from infected B6 mice on day 2 post-infection. CCR2hi, CCR2int and CCR2lo subsets within the MHC IIhigh CD11c+ population I were FACS-sorted and co-cultured with CD4+ Hector T cells for 1 day. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were then analyzed for CD69 expression. Representative FACS plots are shown, Data are representative of 2 independent experiments.
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Show All Figures
getmorefigures.php?uid=PMC4591991&req=5

ppat.1005164.g006: Flt3L-dependent migratory DCs and monocyte-derived DCs both present C. albicans derived antigen.(A) Cervical lymph node cells of B6 or Flt3l-/- mice were analyzed on day 2 post-infection. Representative FACS plots from individual mice and quantification of MHC IIhigh CD11c+ (population I), MHC II+ CD11chigh (population II) and MHC II+ CD11c+ cells (population III) from one of two independent experiments are shown. (B, C) Cervical lymph node cells were isolated from naïve B6 mice and from infected B6 and Flt3l-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were analyzed for CD69 expression after 1 day (B) and for proliferation after 4 days, respectively (C). Representative plots are shown on the left, summary of data from individual mice with mean + SD are shown on the right. Each symbol represents one mouse. Data are representative of 2 independent experiments. (D) As in B, but MHC IIhigh CD11c+ migratory DCs were isolated by FACS-sorting from the cervical lymph nodes of infected B6 and Flt3l-/- mice. (E—F) CCR2+ CD11b+ cells in the cervical lymph nodes of naïve and infected B6 mice on day 2 post-infection were analyzed for the expression of MHC II and CD11c (D). Three distinct subsets of CCR2+ CD11b+ cells (MHC IIlo/int Ly6C+ (Q1), MHC IIint Ly6C- (Q2) and MHC IIhigh Ly6C- (Q3)) from infected mice were further analyzed for the expression of CCR2 and CD11c (E). Representative FACS plots are shown. (G) Cervical lymph nodes were isolated from infected B6 mice on day 2 post-infection. CCR2hi, CCR2int and CCR2lo subsets within the MHC IIhigh CD11c+ population I were FACS-sorted and co-cultured with CD4+ Hector T cells for 1 day. Thy1.1+ CD3+ CD4+ TCRVα2+ cells were then analyzed for CD69 expression. Representative FACS plots are shown, Data are representative of 2 independent experiments.
Mentions: The large Langerin−CD103– subset of migratory DCs in the cervical lymph node appeared to be phenotypically homogeneous for all markers analyzed (Fig 4B). However, the subset may still comprise phylogenetically and functionally distinct cell types, including Flt3-dependent conventional DCs and monocyte-derived DCs that are Flt3-independent but dependent on Csf1R signaling for differentiation from inflammatory monocyte [13]. Consistent with this notion, we found that in Flt3l-/- mice, migratory MHC IIhi CD11c+ DCs, which include the large population of Langerin−CD103– cells, were strongly reduced in Flt3l-/- mice compared to B6 mice (Fig 6A). Furthermore, the CD11b+ cervical lymph node cells isolated from infected Flt3l-/- mice were strongly impaired in their ability to induce CD69 upregulation and proliferation of Hector T cells in vitro (Fig 6B and 6C). This became also clear when the APC fraction was purified by FACS sorting: antigen presentation by Flt3L-dependent migratory DCs strongly promoted the activation of CD4+ Hector T cells in vitro, it was however not essential for the response (Fig 6D), indicating that Flt3L-independent migratory DCs are also involved. Thus, Flt3L-dependent migratory DCs appear to be an important source of antigen in cervical lymph nodes for T cell activation in response to C. albicans oral infection.

Bottom Line: The events regulating T cell activation and differentiation toward effector fates in response to fungal invasion in different tissues are poorly understood.This highlights the functional compartmentalization of specific DC subsets in different tissues.These data provide important new insights to our understanding of tissue-specific antifungal immunity.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microbiology, ETH Zürich, Zürich, Switzerland.

ABSTRACT
Candida spp. can cause severe and chronic mucocutaneous and systemic infections in immunocompromised individuals. Protection from mucocutaneous candidiasis depends on T helper cells, in particular those secreting IL-17. The events regulating T cell activation and differentiation toward effector fates in response to fungal invasion in different tissues are poorly understood. Here we generated a Candida-specific TCR transgenic mouse reactive to a novel endogenous antigen that is conserved in multiple distant species of Candida, including the clinically highly relevant C. albicans and C. glabrata. Using TCR transgenic T cells in combination with an experimental model of oropharyngeal candidiasis (OPC) we investigated antigen presentation and Th17 priming by different subsets of dendritic cells (DCs) present in the infected oral mucosa. Candida-derived endogenous antigen accesses the draining lymph nodes and is directly presented by migratory DCs. Tissue-resident Flt3L-dependent DCs and CCR2-dependent monocyte-derived DCs collaborate in antigen presentation and T cell priming during OPC. In contrast, Langerhans cells, which are also present in the oral mucosa and have been shown to prime Th17 cells in the skin, are not required for induction of the Candida-specific T cell response upon oral challenge. This highlights the functional compartmentalization of specific DC subsets in different tissues. These data provide important new insights to our understanding of tissue-specific antifungal immunity.

No MeSH data available.


Related in: MedlinePlus