Limits...
The transcription factor lymphoid enhancer factor 1 controls invariant natural killer T cell expansion and Th2-type effector differentiation.

Carr T, Krishnamoorthy V, Yu S, Xue HH, Kee BL, Verykokakis M - J. Exp. Med. (2015)

Bottom Line: These cells acquire multiple effector fates during their thymic development that parallel those of CD4(+) T helper cells.LEF1 also directly augments expression of the effector fate-specifying transcription factor GATA3, thus promoting the development of Th2-like effector iNKT cells that produce IL-4, including those that also produce interferon-γ.Our data reveal LEF1 as a central regulator of iNKT cell number and Th2-type effector differentiation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Committee on Immunology, Committee on Molecular Pathogenesis and Molecular Medicine, and Department of Pathology, The University of Chicago, Chicago, IL 60637.

Show MeSH

Related in: MedlinePlus

LEF1 promoted development of iNKT cells that produce IL-4 or IL-4 plus IFNγ. (A) Expression of IL-4 and IFNγ in PLZFhi and PLZFlo iNKT cells was determined by flow cytometry, after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. (B) Expression of IL-4, IFNγ, or IL-17A in thymic Tetr+TCRβ+ iNKT cells from control and Lef1Δ/Δ mice after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. Plots are representative of four to six independent experiments. (C) The mean percentage of control and Lef1Δ/Δ iNKT cells that expressed IL-4, IFNγ, IL-4 plus IFNγ, or IL-17A. Data were averaged from six independent experiments. (D) Expression of CD8α and EOMES in TCRβ+CD8α+ thymocytes was determined by flow cytometry. (E) The mean percentage of EOMES+ cells among TCRβ+CD8α+ thymocytes from four independent experiments. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4419352&req=5

fig4: LEF1 promoted development of iNKT cells that produce IL-4 or IL-4 plus IFNγ. (A) Expression of IL-4 and IFNγ in PLZFhi and PLZFlo iNKT cells was determined by flow cytometry, after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. (B) Expression of IL-4, IFNγ, or IL-17A in thymic Tetr+TCRβ+ iNKT cells from control and Lef1Δ/Δ mice after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. Plots are representative of four to six independent experiments. (C) The mean percentage of control and Lef1Δ/Δ iNKT cells that expressed IL-4, IFNγ, IL-4 plus IFNγ, or IL-17A. Data were averaged from six independent experiments. (D) Expression of CD8α and EOMES in TCRβ+CD8α+ thymocytes was determined by flow cytometry. (E) The mean percentage of EOMES+ cells among TCRβ+CD8α+ thymocytes from four independent experiments. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Mentions: The cytokine production profiles for the iNKT cell effector fates are substantially more flexible than in their CD4+ Th cell counterparts. Dual production of IL-4 and IFNγ is a hallmark of iNKT cells that has been attributed to their high expression of PLZF (Savage et al., 2008). Indeed, some PLZFhi iNKT cells produce both IFNγ and IL-4 after in vitro stimulation with PMA and ionomycin, whereas PLZFlo iNKT cells produced IFNγ exclusively (Fig. 4 A). Given that the majority of PLZFhi cells are iNKT2 cells, we anticipated that LEF1 would be required for the development of the dual IL-4 plus IFNγ–producing population. Indeed, in vitro stimulation of total iNKT cells with PMA and ionomycin revealed that a lower frequency of Lef1Δ/Δ iNKT cells produced the Th2 cytokine IL-4 and the frequency of IL-4 plus IFNγ producers was also reduced (Fig. 4, B and C). In contrast, the frequency of Lef1Δ/Δ iNKT cells producing IFNγ only was not different from control mice, and IL-17A–producing cells were increased (Fig. 4, B and C). Therefore, LEF1 was also required for the development of functional iNKT2 cells that produced IL-4 and IFNγ.


The transcription factor lymphoid enhancer factor 1 controls invariant natural killer T cell expansion and Th2-type effector differentiation.

Carr T, Krishnamoorthy V, Yu S, Xue HH, Kee BL, Verykokakis M - J. Exp. Med. (2015)

LEF1 promoted development of iNKT cells that produce IL-4 or IL-4 plus IFNγ. (A) Expression of IL-4 and IFNγ in PLZFhi and PLZFlo iNKT cells was determined by flow cytometry, after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. (B) Expression of IL-4, IFNγ, or IL-17A in thymic Tetr+TCRβ+ iNKT cells from control and Lef1Δ/Δ mice after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. Plots are representative of four to six independent experiments. (C) The mean percentage of control and Lef1Δ/Δ iNKT cells that expressed IL-4, IFNγ, IL-4 plus IFNγ, or IL-17A. Data were averaged from six independent experiments. (D) Expression of CD8α and EOMES in TCRβ+CD8α+ thymocytes was determined by flow cytometry. (E) The mean percentage of EOMES+ cells among TCRβ+CD8α+ thymocytes from four independent experiments. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4419352&req=5

fig4: LEF1 promoted development of iNKT cells that produce IL-4 or IL-4 plus IFNγ. (A) Expression of IL-4 and IFNγ in PLZFhi and PLZFlo iNKT cells was determined by flow cytometry, after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. (B) Expression of IL-4, IFNγ, or IL-17A in thymic Tetr+TCRβ+ iNKT cells from control and Lef1Δ/Δ mice after a 4-h in vitro stimulation of thymocytes with PMA and ionomycin. Plots are representative of four to six independent experiments. (C) The mean percentage of control and Lef1Δ/Δ iNKT cells that expressed IL-4, IFNγ, IL-4 plus IFNγ, or IL-17A. Data were averaged from six independent experiments. (D) Expression of CD8α and EOMES in TCRβ+CD8α+ thymocytes was determined by flow cytometry. (E) The mean percentage of EOMES+ cells among TCRβ+CD8α+ thymocytes from four independent experiments. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Mentions: The cytokine production profiles for the iNKT cell effector fates are substantially more flexible than in their CD4+ Th cell counterparts. Dual production of IL-4 and IFNγ is a hallmark of iNKT cells that has been attributed to their high expression of PLZF (Savage et al., 2008). Indeed, some PLZFhi iNKT cells produce both IFNγ and IL-4 after in vitro stimulation with PMA and ionomycin, whereas PLZFlo iNKT cells produced IFNγ exclusively (Fig. 4 A). Given that the majority of PLZFhi cells are iNKT2 cells, we anticipated that LEF1 would be required for the development of the dual IL-4 plus IFNγ–producing population. Indeed, in vitro stimulation of total iNKT cells with PMA and ionomycin revealed that a lower frequency of Lef1Δ/Δ iNKT cells produced the Th2 cytokine IL-4 and the frequency of IL-4 plus IFNγ producers was also reduced (Fig. 4, B and C). In contrast, the frequency of Lef1Δ/Δ iNKT cells producing IFNγ only was not different from control mice, and IL-17A–producing cells were increased (Fig. 4, B and C). Therefore, LEF1 was also required for the development of functional iNKT2 cells that produced IL-4 and IFNγ.

Bottom Line: These cells acquire multiple effector fates during their thymic development that parallel those of CD4(+) T helper cells.LEF1 also directly augments expression of the effector fate-specifying transcription factor GATA3, thus promoting the development of Th2-like effector iNKT cells that produce IL-4, including those that also produce interferon-γ.Our data reveal LEF1 as a central regulator of iNKT cell number and Th2-type effector differentiation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Committee on Immunology, Committee on Molecular Pathogenesis and Molecular Medicine, and Department of Pathology, The University of Chicago, Chicago, IL 60637.

Show MeSH
Related in: MedlinePlus