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A unique DNA methylation signature defines a population of IFN-γ/IL-4 double-positive T cells during helminth infection.

Deaton AM, Cook PC, De Sousa D, Phythian-Adams AT, Bird A, MacDonald AS - Eur. J. Immunol. (2014)

Bottom Line: These IFN-γ(+) IL-4(+) cells showed differences in DNA methylation at the Ifng and Il4 loci when compared with IFN-γ(+) IL-4(-) (Th1) and IFN-γ(-) IL-4(+) (Th2) cells, demonstrating that they represent a distinct effector cell population.DNA methylation at this region correlated with decreased Gata3 levels, suggesting a possible role in controlling Gata3 expression.These data provide important insight into the molecular mechanisms behind the co-existence of Th1 and Th2 characteristics.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, UK.

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IFN-γ+IL-4+ cells are generated during Schistosoma mansoni infection. Splenocytes were isolated from infected mice taken from the same experiment (Wk 8) or uninfected age-matched controls (N). (A) The proportion and number of splenic CD4+ T cells was assessed by flow cytometry. Each symbol represents an individual animal and the mean of each sample group is shown as a horizontal line. Statistical significance was assessed using a Student's t-test and data are representative of two independent experiments. (B) Purified splenic CD4+ T cells were stimulated with PMA, ionomycin, and GolgiStop. FACS plots show intracellular staining of IFN-γ and IL-4. Dead cells, doublets and TCR-β−CD4− cells were excluded prior to assessing cytokine expression (Supporting Information Fig. 1). Plots are representative of five independent infections where either pools of spleens were analyzed or animals were analyzed separately. Results were similar in both cases. The percentage of splenic CD4+ cells expressing IFN-γ alone, both IFN-γ and IL-4, or IL-4 alone for individual mice taken from the same experiment are plotted on the right hand side of the figure, significance was assessed using two-way ANOVA. *p < 0.05, **p < 0.01 and ****p < 0.0001.
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fig01: IFN-γ+IL-4+ cells are generated during Schistosoma mansoni infection. Splenocytes were isolated from infected mice taken from the same experiment (Wk 8) or uninfected age-matched controls (N). (A) The proportion and number of splenic CD4+ T cells was assessed by flow cytometry. Each symbol represents an individual animal and the mean of each sample group is shown as a horizontal line. Statistical significance was assessed using a Student's t-test and data are representative of two independent experiments. (B) Purified splenic CD4+ T cells were stimulated with PMA, ionomycin, and GolgiStop. FACS plots show intracellular staining of IFN-γ and IL-4. Dead cells, doublets and TCR-β−CD4− cells were excluded prior to assessing cytokine expression (Supporting Information Fig. 1). Plots are representative of five independent infections where either pools of spleens were analyzed or animals were analyzed separately. Results were similar in both cases. The percentage of splenic CD4+ cells expressing IFN-γ alone, both IFN-γ and IL-4, or IL-4 alone for individual mice taken from the same experiment are plotted on the right hand side of the figure, significance was assessed using two-way ANOVA. *p < 0.05, **p < 0.01 and ****p < 0.0001.

Mentions: In order to examine DNA methylation in an in vivo infection setting we isolated splenic CD4+ T cells from mice that had been infected with S. mansoni for 8 weeks and from age-matched uninfected controls (Fig.1A). A marked proportion of CD4+ T cells displayed properties of both Th1 and Th2 cells in that they simultaneously made both IFN-γ and IL-4 8 (Fig.1B and Supporting Information Fig. 1). Conventional IFN-γ+IL-4− Th1 cells and IFN-γ−IL-4+ Th2 cells were also present, consistent with previous reports 18,19 and CD4+ T cells from uninfected mice showed significantly less expression of IFN-γ or IL-4 (Fig.1B). IFN-γ+IL-4+ cells were observed in five separate S. mansoni infections with the proportion varying from approximately 2–9% of CD4+ T cells (data not shown), demonstrating that IFN-γ+IL-4+ cells can be found in the spleen in a Th2-dominated infection setting.


A unique DNA methylation signature defines a population of IFN-γ/IL-4 double-positive T cells during helminth infection.

Deaton AM, Cook PC, De Sousa D, Phythian-Adams AT, Bird A, MacDonald AS - Eur. J. Immunol. (2014)

IFN-γ+IL-4+ cells are generated during Schistosoma mansoni infection. Splenocytes were isolated from infected mice taken from the same experiment (Wk 8) or uninfected age-matched controls (N). (A) The proportion and number of splenic CD4+ T cells was assessed by flow cytometry. Each symbol represents an individual animal and the mean of each sample group is shown as a horizontal line. Statistical significance was assessed using a Student's t-test and data are representative of two independent experiments. (B) Purified splenic CD4+ T cells were stimulated with PMA, ionomycin, and GolgiStop. FACS plots show intracellular staining of IFN-γ and IL-4. Dead cells, doublets and TCR-β−CD4− cells were excluded prior to assessing cytokine expression (Supporting Information Fig. 1). Plots are representative of five independent infections where either pools of spleens were analyzed or animals were analyzed separately. Results were similar in both cases. The percentage of splenic CD4+ cells expressing IFN-γ alone, both IFN-γ and IL-4, or IL-4 alone for individual mice taken from the same experiment are plotted on the right hand side of the figure, significance was assessed using two-way ANOVA. *p < 0.05, **p < 0.01 and ****p < 0.0001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4231227&req=5

fig01: IFN-γ+IL-4+ cells are generated during Schistosoma mansoni infection. Splenocytes were isolated from infected mice taken from the same experiment (Wk 8) or uninfected age-matched controls (N). (A) The proportion and number of splenic CD4+ T cells was assessed by flow cytometry. Each symbol represents an individual animal and the mean of each sample group is shown as a horizontal line. Statistical significance was assessed using a Student's t-test and data are representative of two independent experiments. (B) Purified splenic CD4+ T cells were stimulated with PMA, ionomycin, and GolgiStop. FACS plots show intracellular staining of IFN-γ and IL-4. Dead cells, doublets and TCR-β−CD4− cells were excluded prior to assessing cytokine expression (Supporting Information Fig. 1). Plots are representative of five independent infections where either pools of spleens were analyzed or animals were analyzed separately. Results were similar in both cases. The percentage of splenic CD4+ cells expressing IFN-γ alone, both IFN-γ and IL-4, or IL-4 alone for individual mice taken from the same experiment are plotted on the right hand side of the figure, significance was assessed using two-way ANOVA. *p < 0.05, **p < 0.01 and ****p < 0.0001.
Mentions: In order to examine DNA methylation in an in vivo infection setting we isolated splenic CD4+ T cells from mice that had been infected with S. mansoni for 8 weeks and from age-matched uninfected controls (Fig.1A). A marked proportion of CD4+ T cells displayed properties of both Th1 and Th2 cells in that they simultaneously made both IFN-γ and IL-4 8 (Fig.1B and Supporting Information Fig. 1). Conventional IFN-γ+IL-4− Th1 cells and IFN-γ−IL-4+ Th2 cells were also present, consistent with previous reports 18,19 and CD4+ T cells from uninfected mice showed significantly less expression of IFN-γ or IL-4 (Fig.1B). IFN-γ+IL-4+ cells were observed in five separate S. mansoni infections with the proportion varying from approximately 2–9% of CD4+ T cells (data not shown), demonstrating that IFN-γ+IL-4+ cells can be found in the spleen in a Th2-dominated infection setting.

Bottom Line: These IFN-γ(+) IL-4(+) cells showed differences in DNA methylation at the Ifng and Il4 loci when compared with IFN-γ(+) IL-4(-) (Th1) and IFN-γ(-) IL-4(+) (Th2) cells, demonstrating that they represent a distinct effector cell population.DNA methylation at this region correlated with decreased Gata3 levels, suggesting a possible role in controlling Gata3 expression.These data provide important insight into the molecular mechanisms behind the co-existence of Th1 and Th2 characteristics.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, UK.

Show MeSH
Related in: MedlinePlus