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CD161 expression characterizes a subpopulation of human regulatory T cells that produces IL-17 in a STAT3-dependent manner.

Afzali B, Mitchell PJ, Edozie FC, Povoleri GA, Dowson SE, Demandt L, Walter G, Canavan JB, Scotta C, Menon B, Chana PS, Khamri W, Kordasti SY, Heck S, Grimbacher B, Tree T, Cope AP, Taams LS, Lechler RI, John S, Lombardi G - Eur. J. Immunol. (2013)

Bottom Line: Importantly, we find that IL-17 production is STAT3 dependent, with Treg cells from patients with STAT3 mutations unable to make IL-17.Finally, we show that CD161(+) population III Treg cells accumulate in inflamed joints of patients with inflammatory arthritis and are the predominant IL-17-producing Treg-cell population at these sites.As IL-17 production from this Treg-cell subpopulation is not accompanied by a loss of regulatory function, in the context of cell therapy, exclusion of these cells from the cell product may not be necessary.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Centre for Transplantation, King's College London, King's Health Partners, Guy's Hospital, London, UK. susan.john@kcl.ac.uk

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Population III CD161+ Treg cells express less FOXP3 but are as suppressive as other Treg-cell subpopulations. (A) A representative histogram of FOXP3 expression in freshly isolated Treg-cell populations III CD161− (solid line) and III CD161+ (dashed line) relative to isotype control (shaded) is shown. (B) The percentage of FOXP3+ cells is also shown as mean + SD of four independent donors * < 0.05, paired t-test. (C, D) The suppressive effects of freshly isolated population I CD161−, population III CD161−, and population III CD161+ cells on proliferation of autologous CFSE-labeled Teff cells are shown as (C) a representative flow cytometry example and (D) mean + SD of five samples pooled from five independent experiments. (E, F) The suppressive effects of population I CD161−, population III CD161− and population III CD161+, Treg cells on (E) IFN-γ and (F) IL-2 production by autologous Teff cells were determined by supernatant cytokine concentrations (left) and percentage suppression (right) and expressed as mean + SD of two samples pooled from two independent experiments. (G) The suppressive function of population III CD161+ Treg cells after culture under IL-17-inducing conditions was determined by inhibition of CFSE dilution and is shown as a representative example (left) and mean +SD of two donors (right). (H) The concentration of IFN-γ (top left) and IL-2 (bottom left) and the percentage suppression of each cytokine (right) are shown as mean + SD of two samples pooled from two independent experiments. *p < 0.05, one-way ANOVA.
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fig03: Population III CD161+ Treg cells express less FOXP3 but are as suppressive as other Treg-cell subpopulations. (A) A representative histogram of FOXP3 expression in freshly isolated Treg-cell populations III CD161− (solid line) and III CD161+ (dashed line) relative to isotype control (shaded) is shown. (B) The percentage of FOXP3+ cells is also shown as mean + SD of four independent donors * < 0.05, paired t-test. (C, D) The suppressive effects of freshly isolated population I CD161−, population III CD161−, and population III CD161+ cells on proliferation of autologous CFSE-labeled Teff cells are shown as (C) a representative flow cytometry example and (D) mean + SD of five samples pooled from five independent experiments. (E, F) The suppressive effects of population I CD161−, population III CD161− and population III CD161+, Treg cells on (E) IFN-γ and (F) IL-2 production by autologous Teff cells were determined by supernatant cytokine concentrations (left) and percentage suppression (right) and expressed as mean + SD of two samples pooled from two independent experiments. (G) The suppressive function of population III CD161+ Treg cells after culture under IL-17-inducing conditions was determined by inhibition of CFSE dilution and is shown as a representative example (left) and mean +SD of two donors (right). (H) The concentration of IFN-γ (top left) and IL-2 (bottom left) and the percentage suppression of each cytokine (right) are shown as mean + SD of two samples pooled from two independent experiments. *p < 0.05, one-way ANOVA.

Mentions: To further characterize the population of interest, FOXP3 expression was evaluated. Population III CD161+ Treg cells expressed less FOXP3 protein in comparison with population III CD161− cells (Fig. 3A and B). Given the lower expression of FOXP3, and previous suggestions that population III is nonsuppressive [32], we tested their capacity to suppress proliferation of auto-logous CFSE-labeled Teff cells. We found that suppressive function in population III CD161+ Treg cells was similar to other Treg-cell populations (Fig. 3C and D). Likewise, their ability to suppress production of IFN-γ and IL-2 production from Teff cells was comparable to those of other Treg subpopulations (Fig. 3E and F). These data demonstrate that population III CD161+ cells have regulatory properties and further confirmed that these are not contaminating Teff cells of the Th17 or Th1 lineage.


CD161 expression characterizes a subpopulation of human regulatory T cells that produces IL-17 in a STAT3-dependent manner.

Afzali B, Mitchell PJ, Edozie FC, Povoleri GA, Dowson SE, Demandt L, Walter G, Canavan JB, Scotta C, Menon B, Chana PS, Khamri W, Kordasti SY, Heck S, Grimbacher B, Tree T, Cope AP, Taams LS, Lechler RI, John S, Lombardi G - Eur. J. Immunol. (2013)

Population III CD161+ Treg cells express less FOXP3 but are as suppressive as other Treg-cell subpopulations. (A) A representative histogram of FOXP3 expression in freshly isolated Treg-cell populations III CD161− (solid line) and III CD161+ (dashed line) relative to isotype control (shaded) is shown. (B) The percentage of FOXP3+ cells is also shown as mean + SD of four independent donors * < 0.05, paired t-test. (C, D) The suppressive effects of freshly isolated population I CD161−, population III CD161−, and population III CD161+ cells on proliferation of autologous CFSE-labeled Teff cells are shown as (C) a representative flow cytometry example and (D) mean + SD of five samples pooled from five independent experiments. (E, F) The suppressive effects of population I CD161−, population III CD161− and population III CD161+, Treg cells on (E) IFN-γ and (F) IL-2 production by autologous Teff cells were determined by supernatant cytokine concentrations (left) and percentage suppression (right) and expressed as mean + SD of two samples pooled from two independent experiments. (G) The suppressive function of population III CD161+ Treg cells after culture under IL-17-inducing conditions was determined by inhibition of CFSE dilution and is shown as a representative example (left) and mean +SD of two donors (right). (H) The concentration of IFN-γ (top left) and IL-2 (bottom left) and the percentage suppression of each cytokine (right) are shown as mean + SD of two samples pooled from two independent experiments. *p < 0.05, one-way ANOVA.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3815561&req=5

fig03: Population III CD161+ Treg cells express less FOXP3 but are as suppressive as other Treg-cell subpopulations. (A) A representative histogram of FOXP3 expression in freshly isolated Treg-cell populations III CD161− (solid line) and III CD161+ (dashed line) relative to isotype control (shaded) is shown. (B) The percentage of FOXP3+ cells is also shown as mean + SD of four independent donors * < 0.05, paired t-test. (C, D) The suppressive effects of freshly isolated population I CD161−, population III CD161−, and population III CD161+ cells on proliferation of autologous CFSE-labeled Teff cells are shown as (C) a representative flow cytometry example and (D) mean + SD of five samples pooled from five independent experiments. (E, F) The suppressive effects of population I CD161−, population III CD161− and population III CD161+, Treg cells on (E) IFN-γ and (F) IL-2 production by autologous Teff cells were determined by supernatant cytokine concentrations (left) and percentage suppression (right) and expressed as mean + SD of two samples pooled from two independent experiments. (G) The suppressive function of population III CD161+ Treg cells after culture under IL-17-inducing conditions was determined by inhibition of CFSE dilution and is shown as a representative example (left) and mean +SD of two donors (right). (H) The concentration of IFN-γ (top left) and IL-2 (bottom left) and the percentage suppression of each cytokine (right) are shown as mean + SD of two samples pooled from two independent experiments. *p < 0.05, one-way ANOVA.
Mentions: To further characterize the population of interest, FOXP3 expression was evaluated. Population III CD161+ Treg cells expressed less FOXP3 protein in comparison with population III CD161− cells (Fig. 3A and B). Given the lower expression of FOXP3, and previous suggestions that population III is nonsuppressive [32], we tested their capacity to suppress proliferation of auto-logous CFSE-labeled Teff cells. We found that suppressive function in population III CD161+ Treg cells was similar to other Treg-cell populations (Fig. 3C and D). Likewise, their ability to suppress production of IFN-γ and IL-2 production from Teff cells was comparable to those of other Treg subpopulations (Fig. 3E and F). These data demonstrate that population III CD161+ cells have regulatory properties and further confirmed that these are not contaminating Teff cells of the Th17 or Th1 lineage.

Bottom Line: Importantly, we find that IL-17 production is STAT3 dependent, with Treg cells from patients with STAT3 mutations unable to make IL-17.Finally, we show that CD161(+) population III Treg cells accumulate in inflamed joints of patients with inflammatory arthritis and are the predominant IL-17-producing Treg-cell population at these sites.As IL-17 production from this Treg-cell subpopulation is not accompanied by a loss of regulatory function, in the context of cell therapy, exclusion of these cells from the cell product may not be necessary.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Centre for Transplantation, King's College London, King's Health Partners, Guy's Hospital, London, UK. susan.john@kcl.ac.uk

Show MeSH
Related in: MedlinePlus