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Fonsecaea pedrosoi-induced Th17-cell differentiation in mice is fostered by Dectin-2 and suppressed by Mincle recognition.

Wüthrich M, Wang H, Li M, Lerksuthirat T, Hardison SE, Brown GD, Klein B - Eur. J. Immunol. (2015)

Bottom Line: Here, we investigated whether costimulation by TLR agonists fosters the development of adaptive immune responses, by examining the development of fungus-specific T cells.Subcutaneous infection of mice with F. pedrosoi spores induced the activation, expansion, and differentiation of Ag-specific CD4(+) T cells but TLR costimulation did not further augment these T-cell responses.The Dectin-2/FcRγ/Card9 signaling pathway promoted the differentiation of fungus-specific CD4(+) T cells into Th17 cells, whereas Mincle inhibited the development of this T-helper subset in infected mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, University of Wisconsin-Madison, WI, USA.

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F. pedrosoi spores trigger Dectin‐1 and Dectin‐2 signaling and IL‐6 production. (A) BWZ cells and a subline expressing Dectin‐1‐CD3ζ (Dectin‐1), as well as B3Z cells expressing Dectin‐2, Mincle, FcRγ chain, Dectin‐2 + FcRγ, MCL + FcRγ, Dectin‐2 + MCL + FcRγ, Mincle + FcRγ or Mincle + MCL + FcRγ were stimulated with live F. pedrosoi spores. After 18 h, lacZ activity was measured using a colorimetric assay and expressed as OD 560/620 values. Data are expressed as the mean ± SD of duplicate wells. Data are from single experiments representative of three independent experiments. (B) Bone marrow derived dendritic cells (BMDC) were cocultured with live F. pedrosoi spores for 24 h and IL‐6 was detected in the supernatants by ELISA. Numbers above histogram bars indicate the n‐fold change versus the corresponding wild‐type control. Data are from a single experiment representative of two independent experiments.
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eji3393-fig-0002: F. pedrosoi spores trigger Dectin‐1 and Dectin‐2 signaling and IL‐6 production. (A) BWZ cells and a subline expressing Dectin‐1‐CD3ζ (Dectin‐1), as well as B3Z cells expressing Dectin‐2, Mincle, FcRγ chain, Dectin‐2 + FcRγ, MCL + FcRγ, Dectin‐2 + MCL + FcRγ, Mincle + FcRγ or Mincle + MCL + FcRγ were stimulated with live F. pedrosoi spores. After 18 h, lacZ activity was measured using a colorimetric assay and expressed as OD 560/620 values. Data are expressed as the mean ± SD of duplicate wells. Data are from single experiments representative of three independent experiments. (B) Bone marrow derived dendritic cells (BMDC) were cocultured with live F. pedrosoi spores for 24 h and IL‐6 was detected in the supernatants by ELISA. Numbers above histogram bars indicate the n‐fold change versus the corresponding wild‐type control. Data are from a single experiment representative of two independent experiments.

Mentions: Innate recognition of F. pedrosoi and F. monophora requires Mincle on murine macrophages and human dendritic cells 8, 14. Since F. pedrosoi spores triggered an adaptive T‐cell response in vivo, we sought to investigate whether surface expressed CLRs are involved in the innate recognition of the fungal spores. We cocultured live F. pedrosoi spores with CLR transformed B3Z T‐cell hybridoma cells expressing an NFAT‐lacZ β‐galatosidase reporter of ITAM signaling 15. In response to spore stimulation, lacZ activity was increased in reporter cells expressing Dectin‐1 and Dectin‐2 and to a lesser extent MCL (Dectin‐3, Clecsf8, or Clec4d) and Mincle, but not in cells expressing FcRγ only (Fig. 2). Since MCL has been shown to hetero‐dimerize with Dectin‐2 16 and induce coexpression of Mincle on the cell surface 17 we also measured lacZ activity in reporter cells coexpressing MCL/Dectin‐2 and MCL/Mincle. Dectin‐2/MCL and Mincle/MCL coexpressing T‐hybridoma cells produced lacZ activity comparable to Dectin‐2 and Mincle single expression cells, respectively, indicating that coexpression of MCL did not increase Dectin‐2 and Mincle induced signaling. Thus, spores recognized by Dectin‐1 and Dectin‐2 induced the strongest signaling activity, whereas Mincle and MCL recognition induced only a weak response.


Fonsecaea pedrosoi-induced Th17-cell differentiation in mice is fostered by Dectin-2 and suppressed by Mincle recognition.

Wüthrich M, Wang H, Li M, Lerksuthirat T, Hardison SE, Brown GD, Klein B - Eur. J. Immunol. (2015)

F. pedrosoi spores trigger Dectin‐1 and Dectin‐2 signaling and IL‐6 production. (A) BWZ cells and a subline expressing Dectin‐1‐CD3ζ (Dectin‐1), as well as B3Z cells expressing Dectin‐2, Mincle, FcRγ chain, Dectin‐2 + FcRγ, MCL + FcRγ, Dectin‐2 + MCL + FcRγ, Mincle + FcRγ or Mincle + MCL + FcRγ were stimulated with live F. pedrosoi spores. After 18 h, lacZ activity was measured using a colorimetric assay and expressed as OD 560/620 values. Data are expressed as the mean ± SD of duplicate wells. Data are from single experiments representative of three independent experiments. (B) Bone marrow derived dendritic cells (BMDC) were cocultured with live F. pedrosoi spores for 24 h and IL‐6 was detected in the supernatants by ELISA. Numbers above histogram bars indicate the n‐fold change versus the corresponding wild‐type control. Data are from a single experiment representative of two independent experiments.
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eji3393-fig-0002: F. pedrosoi spores trigger Dectin‐1 and Dectin‐2 signaling and IL‐6 production. (A) BWZ cells and a subline expressing Dectin‐1‐CD3ζ (Dectin‐1), as well as B3Z cells expressing Dectin‐2, Mincle, FcRγ chain, Dectin‐2 + FcRγ, MCL + FcRγ, Dectin‐2 + MCL + FcRγ, Mincle + FcRγ or Mincle + MCL + FcRγ were stimulated with live F. pedrosoi spores. After 18 h, lacZ activity was measured using a colorimetric assay and expressed as OD 560/620 values. Data are expressed as the mean ± SD of duplicate wells. Data are from single experiments representative of three independent experiments. (B) Bone marrow derived dendritic cells (BMDC) were cocultured with live F. pedrosoi spores for 24 h and IL‐6 was detected in the supernatants by ELISA. Numbers above histogram bars indicate the n‐fold change versus the corresponding wild‐type control. Data are from a single experiment representative of two independent experiments.
Mentions: Innate recognition of F. pedrosoi and F. monophora requires Mincle on murine macrophages and human dendritic cells 8, 14. Since F. pedrosoi spores triggered an adaptive T‐cell response in vivo, we sought to investigate whether surface expressed CLRs are involved in the innate recognition of the fungal spores. We cocultured live F. pedrosoi spores with CLR transformed B3Z T‐cell hybridoma cells expressing an NFAT‐lacZ β‐galatosidase reporter of ITAM signaling 15. In response to spore stimulation, lacZ activity was increased in reporter cells expressing Dectin‐1 and Dectin‐2 and to a lesser extent MCL (Dectin‐3, Clecsf8, or Clec4d) and Mincle, but not in cells expressing FcRγ only (Fig. 2). Since MCL has been shown to hetero‐dimerize with Dectin‐2 16 and induce coexpression of Mincle on the cell surface 17 we also measured lacZ activity in reporter cells coexpressing MCL/Dectin‐2 and MCL/Mincle. Dectin‐2/MCL and Mincle/MCL coexpressing T‐hybridoma cells produced lacZ activity comparable to Dectin‐2 and Mincle single expression cells, respectively, indicating that coexpression of MCL did not increase Dectin‐2 and Mincle induced signaling. Thus, spores recognized by Dectin‐1 and Dectin‐2 induced the strongest signaling activity, whereas Mincle and MCL recognition induced only a weak response.

Bottom Line: Here, we investigated whether costimulation by TLR agonists fosters the development of adaptive immune responses, by examining the development of fungus-specific T cells.Subcutaneous infection of mice with F. pedrosoi spores induced the activation, expansion, and differentiation of Ag-specific CD4(+) T cells but TLR costimulation did not further augment these T-cell responses.The Dectin-2/FcRγ/Card9 signaling pathway promoted the differentiation of fungus-specific CD4(+) T cells into Th17 cells, whereas Mincle inhibited the development of this T-helper subset in infected mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, University of Wisconsin-Madison, WI, USA.

Show MeSH
Related in: MedlinePlus