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Depletion of Regulatory T Cells Induces High Numbers of Dendritic Cells and Unmasks a Subset of Anti-Tumour CD8+CD11c+ PD-1lo Effector T Cells.

Goudin N, Chappert P, Mégret J, Gross DA, Rocha B, Azogui O - PLoS ONE (2016)

Bottom Line: These events are concordant with a substantial increase in CD11b+ resident dendritic cells (DCs) subsets in draining lymph nodes followed by CD8+ DCs.These results indicate that Treg depletion leads to tumour regression by unmasking an increase of DC subsets as a part of a program that optimizes the microenvironment by orchestrating the activation, amplification, and migration of high numbers of fully differentiated CD8+CD11c+PD1lo effector T cells to the tumour sites.They also indicate that a critical pattern of DC subsets correlates with the evolution of the anti-tumour response and provide a template for Treg depletion and DC-based therapy.

View Article: PubMed Central - PubMed

Affiliation: Plateau technique de Cytometrie et d'Imagerie Cellulaire, Structure Fédérative de Recherche Necker, INSERM US 24-CNRS, UMS 3633, Paris, France.

ABSTRACT
Natural regulatory T (Treg) cells interfere with multiple functions, which are crucial for the development of strong anti-tumour responses. In a model of 4T1 mammary carcinoma, depletion of CD25+Tregs results in tumour regression in Balb/c mice, but the mechanisms underlying this process are not fully understood. Here, we show that partial Treg depletion leads to the generation of a particular effector CD8 T cell subset expressing CD11c and low level of PD-1 in tumour draining lymph nodes. These cells have the capacity to migrate into the tumour, to kill DCs, and to locally regulate the anti-tumour response. These events are concordant with a substantial increase in CD11b+ resident dendritic cells (DCs) subsets in draining lymph nodes followed by CD8+ DCs. These results indicate that Treg depletion leads to tumour regression by unmasking an increase of DC subsets as a part of a program that optimizes the microenvironment by orchestrating the activation, amplification, and migration of high numbers of fully differentiated CD8+CD11c+PD1lo effector T cells to the tumour sites. They also indicate that a critical pattern of DC subsets correlates with the evolution of the anti-tumour response and provide a template for Treg depletion and DC-based therapy.

No MeSH data available.


Related in: MedlinePlus

Treg depletion induces high numbers of CD11b+ DCs in tumour-draining LNs.Groups (n = 3) of untreated and PC61-treated mice were injected s.c. with 2x105 4T1 tumour cells and DC subsets later analyses. (A) Gating strategy for DC subsets in normal LNs. Lineage- cells (CD3ε- CD19-B220-) were examined for CD11c and MHC-II expression to quantify lymphoid-resident DCs (CD11chiMHC-IIint) and migratory DCs (CD11cintMHC-IIhi) (center panel). Resident DCs were further gated for the expression of CD11bhi and CD8+ and DEC205+ (right panel). Migratory DCs were further gated into CD103+ DCs (left panel). (B) Numbers of resident DCs, migratory DCs and CD11bhi, CD8+ subsets belonging to the resident DCs in tumour non-draining LNs (nDLN) and draining LNs (DLN) at indicated time after injection of tumour cells. (C) Comparison of the Mean Fluorescence Intensity (MFI) of the MHC II on CD11b+ subsets studied at day 7 in the indicated organs. (D) Histograms show the proliferation of CellTrace violet-labeled naïve CD8 T cells stimulated by DC subsets sorted at day 7 in the indicated organs with the gates shown in (A). Data are representative of two independent experiments for the kinetics and for times at day 7. Bars show mean values ± SEM. *,p<0.05, **,p<0.001, ***,p<0.0001. P values were calculated using Student’s t test.
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pone.0157822.g002: Treg depletion induces high numbers of CD11b+ DCs in tumour-draining LNs.Groups (n = 3) of untreated and PC61-treated mice were injected s.c. with 2x105 4T1 tumour cells and DC subsets later analyses. (A) Gating strategy for DC subsets in normal LNs. Lineage- cells (CD3ε- CD19-B220-) were examined for CD11c and MHC-II expression to quantify lymphoid-resident DCs (CD11chiMHC-IIint) and migratory DCs (CD11cintMHC-IIhi) (center panel). Resident DCs were further gated for the expression of CD11bhi and CD8+ and DEC205+ (right panel). Migratory DCs were further gated into CD103+ DCs (left panel). (B) Numbers of resident DCs, migratory DCs and CD11bhi, CD8+ subsets belonging to the resident DCs in tumour non-draining LNs (nDLN) and draining LNs (DLN) at indicated time after injection of tumour cells. (C) Comparison of the Mean Fluorescence Intensity (MFI) of the MHC II on CD11b+ subsets studied at day 7 in the indicated organs. (D) Histograms show the proliferation of CellTrace violet-labeled naïve CD8 T cells stimulated by DC subsets sorted at day 7 in the indicated organs with the gates shown in (A). Data are representative of two independent experiments for the kinetics and for times at day 7. Bars show mean values ± SEM. *,p<0.05, **,p<0.001, ***,p<0.0001. P values were calculated using Student’s t test.

Mentions: Dendritic cells (DCs) are promising targets for immunotherapy because they play an essential role in initiating and regulating T cell immunity. It has been shown that CD25+Foxp3+ Tregs led to increased numbers of LN-resident DCs [14] but neither the impact of the Treg depletion on the different DC subsets nor the cell-mediated response to tumour cells have not been examined in details. To address this issue in the context of tumour regression versus tumour progression, PC61-treated and untreated mice were injected with 4T1 cells and DC numbers were evaluated by performing a kinetic analysis of each subset in tumour draining LNs (DLNs) and non-draining LNs (nDLNs). The gating strategy to identify these subsets was already described [22, 23] and is shown in Fig 2A. Live lineage- cells were examined for CD11c and MHC II expression characteristic of the main subsets of CD11c+ DCs, i.e, CD11chiMHC IIint LN resident DCs including CD11bhi, CD8+DEC205+ and CD11chiMHC IIhi including CD103+ migratory DCs (mDCs).


Depletion of Regulatory T Cells Induces High Numbers of Dendritic Cells and Unmasks a Subset of Anti-Tumour CD8+CD11c+ PD-1lo Effector T Cells.

Goudin N, Chappert P, Mégret J, Gross DA, Rocha B, Azogui O - PLoS ONE (2016)

Treg depletion induces high numbers of CD11b+ DCs in tumour-draining LNs.Groups (n = 3) of untreated and PC61-treated mice were injected s.c. with 2x105 4T1 tumour cells and DC subsets later analyses. (A) Gating strategy for DC subsets in normal LNs. Lineage- cells (CD3ε- CD19-B220-) were examined for CD11c and MHC-II expression to quantify lymphoid-resident DCs (CD11chiMHC-IIint) and migratory DCs (CD11cintMHC-IIhi) (center panel). Resident DCs were further gated for the expression of CD11bhi and CD8+ and DEC205+ (right panel). Migratory DCs were further gated into CD103+ DCs (left panel). (B) Numbers of resident DCs, migratory DCs and CD11bhi, CD8+ subsets belonging to the resident DCs in tumour non-draining LNs (nDLN) and draining LNs (DLN) at indicated time after injection of tumour cells. (C) Comparison of the Mean Fluorescence Intensity (MFI) of the MHC II on CD11b+ subsets studied at day 7 in the indicated organs. (D) Histograms show the proliferation of CellTrace violet-labeled naïve CD8 T cells stimulated by DC subsets sorted at day 7 in the indicated organs with the gates shown in (A). Data are representative of two independent experiments for the kinetics and for times at day 7. Bars show mean values ± SEM. *,p<0.05, **,p<0.001, ***,p<0.0001. P values were calculated using Student’s t test.
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pone.0157822.g002: Treg depletion induces high numbers of CD11b+ DCs in tumour-draining LNs.Groups (n = 3) of untreated and PC61-treated mice were injected s.c. with 2x105 4T1 tumour cells and DC subsets later analyses. (A) Gating strategy for DC subsets in normal LNs. Lineage- cells (CD3ε- CD19-B220-) were examined for CD11c and MHC-II expression to quantify lymphoid-resident DCs (CD11chiMHC-IIint) and migratory DCs (CD11cintMHC-IIhi) (center panel). Resident DCs were further gated for the expression of CD11bhi and CD8+ and DEC205+ (right panel). Migratory DCs were further gated into CD103+ DCs (left panel). (B) Numbers of resident DCs, migratory DCs and CD11bhi, CD8+ subsets belonging to the resident DCs in tumour non-draining LNs (nDLN) and draining LNs (DLN) at indicated time after injection of tumour cells. (C) Comparison of the Mean Fluorescence Intensity (MFI) of the MHC II on CD11b+ subsets studied at day 7 in the indicated organs. (D) Histograms show the proliferation of CellTrace violet-labeled naïve CD8 T cells stimulated by DC subsets sorted at day 7 in the indicated organs with the gates shown in (A). Data are representative of two independent experiments for the kinetics and for times at day 7. Bars show mean values ± SEM. *,p<0.05, **,p<0.001, ***,p<0.0001. P values were calculated using Student’s t test.
Mentions: Dendritic cells (DCs) are promising targets for immunotherapy because they play an essential role in initiating and regulating T cell immunity. It has been shown that CD25+Foxp3+ Tregs led to increased numbers of LN-resident DCs [14] but neither the impact of the Treg depletion on the different DC subsets nor the cell-mediated response to tumour cells have not been examined in details. To address this issue in the context of tumour regression versus tumour progression, PC61-treated and untreated mice were injected with 4T1 cells and DC numbers were evaluated by performing a kinetic analysis of each subset in tumour draining LNs (DLNs) and non-draining LNs (nDLNs). The gating strategy to identify these subsets was already described [22, 23] and is shown in Fig 2A. Live lineage- cells were examined for CD11c and MHC II expression characteristic of the main subsets of CD11c+ DCs, i.e, CD11chiMHC IIint LN resident DCs including CD11bhi, CD8+DEC205+ and CD11chiMHC IIhi including CD103+ migratory DCs (mDCs).

Bottom Line: These events are concordant with a substantial increase in CD11b+ resident dendritic cells (DCs) subsets in draining lymph nodes followed by CD8+ DCs.These results indicate that Treg depletion leads to tumour regression by unmasking an increase of DC subsets as a part of a program that optimizes the microenvironment by orchestrating the activation, amplification, and migration of high numbers of fully differentiated CD8+CD11c+PD1lo effector T cells to the tumour sites.They also indicate that a critical pattern of DC subsets correlates with the evolution of the anti-tumour response and provide a template for Treg depletion and DC-based therapy.

View Article: PubMed Central - PubMed

Affiliation: Plateau technique de Cytometrie et d'Imagerie Cellulaire, Structure Fédérative de Recherche Necker, INSERM US 24-CNRS, UMS 3633, Paris, France.

ABSTRACT
Natural regulatory T (Treg) cells interfere with multiple functions, which are crucial for the development of strong anti-tumour responses. In a model of 4T1 mammary carcinoma, depletion of CD25+Tregs results in tumour regression in Balb/c mice, but the mechanisms underlying this process are not fully understood. Here, we show that partial Treg depletion leads to the generation of a particular effector CD8 T cell subset expressing CD11c and low level of PD-1 in tumour draining lymph nodes. These cells have the capacity to migrate into the tumour, to kill DCs, and to locally regulate the anti-tumour response. These events are concordant with a substantial increase in CD11b+ resident dendritic cells (DCs) subsets in draining lymph nodes followed by CD8+ DCs. These results indicate that Treg depletion leads to tumour regression by unmasking an increase of DC subsets as a part of a program that optimizes the microenvironment by orchestrating the activation, amplification, and migration of high numbers of fully differentiated CD8+CD11c+PD1lo effector T cells to the tumour sites. They also indicate that a critical pattern of DC subsets correlates with the evolution of the anti-tumour response and provide a template for Treg depletion and DC-based therapy.

No MeSH data available.


Related in: MedlinePlus