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A central role for Notch in effector CD8(+) T cell differentiation.

Backer RA, Helbig C, Gentek R, Kent A, Laidlaw BJ, Dominguez CX, de Souza YS, van Trierum SE, van Beek R, Rimmelzwaan GF, ten Brinke A, Willemsen AM, van Kampen AH, Kaech SM, Blander JM, van Gisbergen K, Amsen D - Nat. Immunol. (2014)

Bottom Line: Activated CD8(+) T cells choose between terminal effector cell (TEC) or memory precursor cell (MPC) fates.We found that the signaling receptor Notch controls this 'choice'.These pathways were subsequently amplified downstream of Notch, creating a positive feedback loop.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Cell Biology and Histology, Academic Medical Center, Amsterdam, the Netherlands. [2] Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam, the Netherlands.

ABSTRACT
Activated CD8(+) T cells choose between terminal effector cell (TEC) or memory precursor cell (MPC) fates. We found that the signaling receptor Notch controls this 'choice'. Notch promoted the differentiation of immediately protective TECs and was correspondingly required for the clearance of acute infection with influenza virus. Notch activated a major portion of the TEC-specific gene-expression program and suppressed the MPC-specific program. Expression of Notch was induced on naive CD8(+) T cells by inflammatory mediators and interleukin 2 (IL-2) via pathways dependent on the metabolic checkpoint kinase mTOR and the transcription factor T-bet. These pathways were subsequently amplified downstream of Notch, creating a positive feedback loop. Notch thus functions as a central hub where information from different sources converges to match effector T cell differentiation to the demands of an infection.

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Notch ligands are expressed on APC during influenza infection. C57BL/6 mice were infected intranasally with A/HK×31 influenza. (a) Five days post infection (p.i.) expression of DLL1, DLL4, Jagged1 and Jagged2 expression on lung APC subsets was measured by flow cytometry. Two main APC subsets in the lung were defined as mDCs (migratory DC, MHCIIhiCD11chi, red gate) or macrophages (Alveolar Mf, SSChiMHCIIintCD11chi, green gate). Filled histograms represent staining with isotype control mAb (Isotype co.), grey lines show Notch ligand expression on APC isolated from non-infected lungs (Uninfected co.), and black lines indicate expression on APC isolated from infected lungs (influenza infected). (b) Mean Fluorescence Intensity of DLL1, DLL4, Jagged1 and Jagged2 on mDC isolated from the lung (left) or isolated from the draining mLN (right). Shown is Δ-MFI (corrected for background staining with isotype control mAb) for non-infected (white bars) and infected mice (black bars). Results represent 2 (uninfected) or 4 (infected) separately processed mice from a representative of 5 experiments. Mean + s.e.m. *P < 0.05, two-tailed t-test.
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Figure 2: Notch ligands are expressed on APC during influenza infection. C57BL/6 mice were infected intranasally with A/HK×31 influenza. (a) Five days post infection (p.i.) expression of DLL1, DLL4, Jagged1 and Jagged2 expression on lung APC subsets was measured by flow cytometry. Two main APC subsets in the lung were defined as mDCs (migratory DC, MHCIIhiCD11chi, red gate) or macrophages (Alveolar Mf, SSChiMHCIIintCD11chi, green gate). Filled histograms represent staining with isotype control mAb (Isotype co.), grey lines show Notch ligand expression on APC isolated from non-infected lungs (Uninfected co.), and black lines indicate expression on APC isolated from infected lungs (influenza infected). (b) Mean Fluorescence Intensity of DLL1, DLL4, Jagged1 and Jagged2 on mDC isolated from the lung (left) or isolated from the draining mLN (right). Shown is Δ-MFI (corrected for background staining with isotype control mAb) for non-infected (white bars) and infected mice (black bars). Results represent 2 (uninfected) or 4 (infected) separately processed mice from a representative of 5 experiments. Mean + s.e.m. *P < 0.05, two-tailed t-test.

Mentions: Two major populations of APCs in lungs are CD11chiMHCIIint alveolar macrophages and CD11chighMHCIIhigh migratory DC (characterized by intermediate and large size, respectively) (Fig. 2a, middle and Supplementary Fig. 3a)33, 34. These migratory DCs (mDCs) were shown to carry influenza antigen to the mediastinal lymph nodes and are responsible for priming naive CD8+ T cells through direct presentation34, 35. Both mDCs and alveolar macrophages expressed undetectable amounts of Dll4 and Jagged2 and low amounts of Dll1 and Jagged1 in lungs from uninfected mice (Fig. 2a, bottom). Expression of Dll1 and Jagged1 was increased on both types of APCs upon infection with influenza virus (Fig. 2a, bottom). This elevated expression persisted on lung-derived migratory DCs (but not lymph node resident macrophages) isolated from the lung-draining mediastinal lymph nodes from infected mice (Fig. 2b and Supplementary Fig. 3b). Yet, Notch ligands were virtually undetectable on other populations of MHCII+ cells present in lungs, even upon infection with influenza (Supplementary Fig. 3c). Therefore, expression of Notch ligands is induced by influenza infection on the APC that prime naive CD8+ T cells, positioning these molecules for a potential role in differentiation of virus specific CD8+ T cells.


A central role for Notch in effector CD8(+) T cell differentiation.

Backer RA, Helbig C, Gentek R, Kent A, Laidlaw BJ, Dominguez CX, de Souza YS, van Trierum SE, van Beek R, Rimmelzwaan GF, ten Brinke A, Willemsen AM, van Kampen AH, Kaech SM, Blander JM, van Gisbergen K, Amsen D - Nat. Immunol. (2014)

Notch ligands are expressed on APC during influenza infection. C57BL/6 mice were infected intranasally with A/HK×31 influenza. (a) Five days post infection (p.i.) expression of DLL1, DLL4, Jagged1 and Jagged2 expression on lung APC subsets was measured by flow cytometry. Two main APC subsets in the lung were defined as mDCs (migratory DC, MHCIIhiCD11chi, red gate) or macrophages (Alveolar Mf, SSChiMHCIIintCD11chi, green gate). Filled histograms represent staining with isotype control mAb (Isotype co.), grey lines show Notch ligand expression on APC isolated from non-infected lungs (Uninfected co.), and black lines indicate expression on APC isolated from infected lungs (influenza infected). (b) Mean Fluorescence Intensity of DLL1, DLL4, Jagged1 and Jagged2 on mDC isolated from the lung (left) or isolated from the draining mLN (right). Shown is Δ-MFI (corrected for background staining with isotype control mAb) for non-infected (white bars) and infected mice (black bars). Results represent 2 (uninfected) or 4 (infected) separately processed mice from a representative of 5 experiments. Mean + s.e.m. *P < 0.05, two-tailed t-test.
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Figure 2: Notch ligands are expressed on APC during influenza infection. C57BL/6 mice were infected intranasally with A/HK×31 influenza. (a) Five days post infection (p.i.) expression of DLL1, DLL4, Jagged1 and Jagged2 expression on lung APC subsets was measured by flow cytometry. Two main APC subsets in the lung were defined as mDCs (migratory DC, MHCIIhiCD11chi, red gate) or macrophages (Alveolar Mf, SSChiMHCIIintCD11chi, green gate). Filled histograms represent staining with isotype control mAb (Isotype co.), grey lines show Notch ligand expression on APC isolated from non-infected lungs (Uninfected co.), and black lines indicate expression on APC isolated from infected lungs (influenza infected). (b) Mean Fluorescence Intensity of DLL1, DLL4, Jagged1 and Jagged2 on mDC isolated from the lung (left) or isolated from the draining mLN (right). Shown is Δ-MFI (corrected for background staining with isotype control mAb) for non-infected (white bars) and infected mice (black bars). Results represent 2 (uninfected) or 4 (infected) separately processed mice from a representative of 5 experiments. Mean + s.e.m. *P < 0.05, two-tailed t-test.
Mentions: Two major populations of APCs in lungs are CD11chiMHCIIint alveolar macrophages and CD11chighMHCIIhigh migratory DC (characterized by intermediate and large size, respectively) (Fig. 2a, middle and Supplementary Fig. 3a)33, 34. These migratory DCs (mDCs) were shown to carry influenza antigen to the mediastinal lymph nodes and are responsible for priming naive CD8+ T cells through direct presentation34, 35. Both mDCs and alveolar macrophages expressed undetectable amounts of Dll4 and Jagged2 and low amounts of Dll1 and Jagged1 in lungs from uninfected mice (Fig. 2a, bottom). Expression of Dll1 and Jagged1 was increased on both types of APCs upon infection with influenza virus (Fig. 2a, bottom). This elevated expression persisted on lung-derived migratory DCs (but not lymph node resident macrophages) isolated from the lung-draining mediastinal lymph nodes from infected mice (Fig. 2b and Supplementary Fig. 3b). Yet, Notch ligands were virtually undetectable on other populations of MHCII+ cells present in lungs, even upon infection with influenza (Supplementary Fig. 3c). Therefore, expression of Notch ligands is induced by influenza infection on the APC that prime naive CD8+ T cells, positioning these molecules for a potential role in differentiation of virus specific CD8+ T cells.

Bottom Line: Activated CD8(+) T cells choose between terminal effector cell (TEC) or memory precursor cell (MPC) fates.We found that the signaling receptor Notch controls this 'choice'.These pathways were subsequently amplified downstream of Notch, creating a positive feedback loop.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Cell Biology and Histology, Academic Medical Center, Amsterdam, the Netherlands. [2] Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam, the Netherlands.

ABSTRACT
Activated CD8(+) T cells choose between terminal effector cell (TEC) or memory precursor cell (MPC) fates. We found that the signaling receptor Notch controls this 'choice'. Notch promoted the differentiation of immediately protective TECs and was correspondingly required for the clearance of acute infection with influenza virus. Notch activated a major portion of the TEC-specific gene-expression program and suppressed the MPC-specific program. Expression of Notch was induced on naive CD8(+) T cells by inflammatory mediators and interleukin 2 (IL-2) via pathways dependent on the metabolic checkpoint kinase mTOR and the transcription factor T-bet. These pathways were subsequently amplified downstream of Notch, creating a positive feedback loop. Notch thus functions as a central hub where information from different sources converges to match effector T cell differentiation to the demands of an infection.

Show MeSH
Related in: MedlinePlus