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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

Transport of C. albicans-derived antigen to the cervical lymph nodes is CCR7-dependent.(A) Cervical lymph node cells that were isolated from naïve or from sublingually infected mice on day 2 post-infection and either enriched for CD11b+ cells or left non-enriched were co-cultured with the C. albicans-specific T cell hybridoma cells. IL-2 production as a read-out for hybridoma activation was quantified by CTLL-2 bioassay. (B) Cervical lymph node cells were isolated from naïve mice or from sublingually infected mice on day 2 p.i., enriched for CD11b+ cells, and co-cultured with CFSE-labeled CD4+ Hector T cells. CFSE-dilution of the Thy1.1+ CD3+ CD4+ Hector cells was analyzed after 4 days. (C) CD11b+ cells were enriched from cervical lymph nodes at different time points after infection as indicated and analyzed for antigen presentation as described in (A). (D—E) cervical lymph node cells were isolated from sublingually infected B6 or Ccr7-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. CD69 expression (D) and CFSE dilution (E) of Thy1.1+ CD3+ CD4+ TCRVα2+ cells was analyzed on day 1 and day4 respectively. In (B), (D) and (E), representative FACS plots are shown on the left; summary of data from individual mice with mean + SD are shown on the right. Data are from individual experiments that are representative of at least two independent experiments each. (F) CD4+ Hector T cells were adoptively transferred into B6 and Ccr7-/- mice one day prior to sublingual infection. Cytokine production by Thy1.1+ CD3+ CD4+ Hector cells in the cervical lymph nodes was analyzed on day 7 post-infection after re-stimulation with DC1940 cells pulsed with pADH1126-140, heat-killed (h.k.) C. albicans or left unpulsed as indicated. Symbols represent individual mice pooled from 2 independent experiments, the mean + SD is indicated.
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ppat.1005164.g003: Transport of C. albicans-derived antigen to the cervical lymph nodes is CCR7-dependent.(A) Cervical lymph node cells that were isolated from naïve or from sublingually infected mice on day 2 post-infection and either enriched for CD11b+ cells or left non-enriched were co-cultured with the C. albicans-specific T cell hybridoma cells. IL-2 production as a read-out for hybridoma activation was quantified by CTLL-2 bioassay. (B) Cervical lymph node cells were isolated from naïve mice or from sublingually infected mice on day 2 p.i., enriched for CD11b+ cells, and co-cultured with CFSE-labeled CD4+ Hector T cells. CFSE-dilution of the Thy1.1+ CD3+ CD4+ Hector cells was analyzed after 4 days. (C) CD11b+ cells were enriched from cervical lymph nodes at different time points after infection as indicated and analyzed for antigen presentation as described in (A). (D—E) cervical lymph node cells were isolated from sublingually infected B6 or Ccr7-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. CD69 expression (D) and CFSE dilution (E) of Thy1.1+ CD3+ CD4+ TCRVα2+ cells was analyzed on day 1 and day4 respectively. In (B), (D) and (E), representative FACS plots are shown on the left; summary of data from individual mice with mean + SD are shown on the right. Data are from individual experiments that are representative of at least two independent experiments each. (F) CD4+ Hector T cells were adoptively transferred into B6 and Ccr7-/- mice one day prior to sublingual infection. Cytokine production by Thy1.1+ CD3+ CD4+ Hector cells in the cervical lymph nodes was analyzed on day 7 post-infection after re-stimulation with DC1940 cells pulsed with pADH1126-140, heat-killed (h.k.) C. albicans or left unpulsed as indicated. Symbols represent individual mice pooled from 2 independent experiments, the mean + SD is indicated.

Mentions: We next asked how in OPC C. albicans-derived antigen can reach the draining lymph node from the site of infection, since the fungus is normally restricted to the keratinized layer of the oral epithelium and does not invade deeper tissues or drain to lymphoid organs in immunocompetent mice [30]. Indeed, we were unable to culture C. albicans from the cervical lymph nodes of infected B6 mice. A likely possibility is that C. albicans-derived antigen accesses draining lymph nodes transported by migratory cells that arrive from the site of infection to the lymph nodes through afferent lymphatics. To test this possibility, we first enriched CD11b+ cells from cervical lymph nodes of C. albicans infected and naïve mice and found that only CD11b+ cells from infected mice could directly and rapidly stimulate the T cell hybridoma 59.8 (Fig 3A) and induce activation and proliferation of Hector T cells in vitro (Fig 3B). Kinetic studies showed that maximal presentation was achieved on day 2 post-infection (Fig 3C). We then enriched CD11b+ cells from cervical lymph nodes of infected B6 and Ccr7–/–mice, in which migration of cells from the periphery to draining lymph nodes is severely impaired [34]. Strikingly, activation of Hector T cells was strongly reduced (Fig 3D and 3E).


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)

Transport of C. albicans-derived antigen to the cervical lymph nodes is CCR7-dependent.(A) Cervical lymph node cells that were isolated from naïve or from sublingually infected mice on day 2 post-infection and either enriched for CD11b+ cells or left non-enriched were co-cultured with the C. albicans-specific T cell hybridoma cells. IL-2 production as a read-out for hybridoma activation was quantified by CTLL-2 bioassay. (B) Cervical lymph node cells were isolated from naïve mice or from sublingually infected mice on day 2 p.i., enriched for CD11b+ cells, and co-cultured with CFSE-labeled CD4+ Hector T cells. CFSE-dilution of the Thy1.1+ CD3+ CD4+ Hector cells was analyzed after 4 days. (C) CD11b+ cells were enriched from cervical lymph nodes at different time points after infection as indicated and analyzed for antigen presentation as described in (A). (D—E) cervical lymph node cells were isolated from sublingually infected B6 or Ccr7-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. CD69 expression (D) and CFSE dilution (E) of Thy1.1+ CD3+ CD4+ TCRVα2+ cells was analyzed on day 1 and day4 respectively. In (B), (D) and (E), representative FACS plots are shown on the left; summary of data from individual mice with mean + SD are shown on the right. Data are from individual experiments that are representative of at least two independent experiments each. (F) CD4+ Hector T cells were adoptively transferred into B6 and Ccr7-/- mice one day prior to sublingual infection. Cytokine production by Thy1.1+ CD3+ CD4+ Hector cells in the cervical lymph nodes was analyzed on day 7 post-infection after re-stimulation with DC1940 cells pulsed with pADH1126-140, heat-killed (h.k.) C. albicans or left unpulsed as indicated. Symbols represent individual mice pooled from 2 independent experiments, the mean + SD is indicated.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4591991&req=5

ppat.1005164.g003: Transport of C. albicans-derived antigen to the cervical lymph nodes is CCR7-dependent.(A) Cervical lymph node cells that were isolated from naïve or from sublingually infected mice on day 2 post-infection and either enriched for CD11b+ cells or left non-enriched were co-cultured with the C. albicans-specific T cell hybridoma cells. IL-2 production as a read-out for hybridoma activation was quantified by CTLL-2 bioassay. (B) Cervical lymph node cells were isolated from naïve mice or from sublingually infected mice on day 2 p.i., enriched for CD11b+ cells, and co-cultured with CFSE-labeled CD4+ Hector T cells. CFSE-dilution of the Thy1.1+ CD3+ CD4+ Hector cells was analyzed after 4 days. (C) CD11b+ cells were enriched from cervical lymph nodes at different time points after infection as indicated and analyzed for antigen presentation as described in (A). (D—E) cervical lymph node cells were isolated from sublingually infected B6 or Ccr7-/- mice on day 2 post-infection, enriched for CD11b+ cells and co-cultured with CD4+ Hector T cells. CD69 expression (D) and CFSE dilution (E) of Thy1.1+ CD3+ CD4+ TCRVα2+ cells was analyzed on day 1 and day4 respectively. In (B), (D) and (E), representative FACS plots are shown on the left; summary of data from individual mice with mean + SD are shown on the right. Data are from individual experiments that are representative of at least two independent experiments each. (F) CD4+ Hector T cells were adoptively transferred into B6 and Ccr7-/- mice one day prior to sublingual infection. Cytokine production by Thy1.1+ CD3+ CD4+ Hector cells in the cervical lymph nodes was analyzed on day 7 post-infection after re-stimulation with DC1940 cells pulsed with pADH1126-140, heat-killed (h.k.) C. albicans or left unpulsed as indicated. Symbols represent individual mice pooled from 2 independent experiments, the mean + SD is indicated.
Mentions: We next asked how in OPC C. albicans-derived antigen can reach the draining lymph node from the site of infection, since the fungus is normally restricted to the keratinized layer of the oral epithelium and does not invade deeper tissues or drain to lymphoid organs in immunocompetent mice [30]. Indeed, we were unable to culture C. albicans from the cervical lymph nodes of infected B6 mice. A likely possibility is that C. albicans-derived antigen accesses draining lymph nodes transported by migratory cells that arrive from the site of infection to the lymph nodes through afferent lymphatics. To test this possibility, we first enriched CD11b+ cells from cervical lymph nodes of C. albicans infected and naïve mice and found that only CD11b+ cells from infected mice could directly and rapidly stimulate the T cell hybridoma 59.8 (Fig 3A) and induce activation and proliferation of Hector T cells in vitro (Fig 3B). Kinetic studies showed that maximal presentation was achieved on day 2 post-infection (Fig 3C). We then enriched CD11b+ cells from cervical lymph nodes of infected B6 and Ccr7–/–mice, in which migration of cells from the periphery to draining lymph nodes is severely impaired [34]. Strikingly, activation of Hector T cells was strongly reduced (Fig 3D and 3E).

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