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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.

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

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LEF1 prevented CD8α expression on peripheral iNKT cells. (A) CD4 and CD8 expression on Tetr+TCRβ+ iNKT cells from the liver of control or Lef1Δ/Δ mice was determined by flow cytometry. (B) The mean number of CD4+, DN, and CD8+ iNKT cells from the indicated tissues in control and Lef1Δ/Δ mice. Data were averaged from at least six independent experiments. (C) CD45.1+CD45.2+ control and CD45.1−CD45.2+Lef1Δ/Δ FACS-sorted LK bone marrow cells were mixed at 1:1 ratio and transferred into lethally irradiated CD45.1+ C57BL/6 mice. 6–8 wk later, reconstituted mice were analyzed by flow cytometry for CD4 and CD8 expression on control or Lef1Δ/Δ iNKT cells from the liver. (D) The mean percentage of the CD4+, DN, and CD8+ liver iNKT cells from competitive BM chimeras. Data are representative of three independent experiments with two to three chimeras each. (E) Th-POK and CD8α expression in splenic iNKT cells from the indicated mice, as determined by flow cytometry. Data are representative of five independent experiments with one to two mice per genotype. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
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fig6: LEF1 prevented CD8α expression on peripheral iNKT cells. (A) CD4 and CD8 expression on Tetr+TCRβ+ iNKT cells from the liver of control or Lef1Δ/Δ mice was determined by flow cytometry. (B) The mean number of CD4+, DN, and CD8+ iNKT cells from the indicated tissues in control and Lef1Δ/Δ mice. Data were averaged from at least six independent experiments. (C) CD45.1+CD45.2+ control and CD45.1−CD45.2+Lef1Δ/Δ FACS-sorted LK bone marrow cells were mixed at 1:1 ratio and transferred into lethally irradiated CD45.1+ C57BL/6 mice. 6–8 wk later, reconstituted mice were analyzed by flow cytometry for CD4 and CD8 expression on control or Lef1Δ/Δ iNKT cells from the liver. (D) The mean percentage of the CD4+, DN, and CD8+ liver iNKT cells from competitive BM chimeras. Data are representative of three independent experiments with two to three chimeras each. (E) Th-POK and CD8α expression in splenic iNKT cells from the indicated mice, as determined by flow cytometry. Data are representative of five independent experiments with one to two mice per genotype. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Mentions: In the liver and spleen of Lef1Δ/Δ mice, the frequency of CD4+ iNKT cells was reduced and a novel population of CD8+ iNKT cells emerged (Fig. 6, A and B). The loss of CD4 and increased expression of CD8 on liver iNKT cells was intrinsic to the Lef1Δ/Δ cells as determined in mixed bone marrow chimeras (Fig. 6, C and D). We examined the expression of Th-POK in Lef1Δ/Δ splenic iNKT cells because Th-POK has been implicated as a GATA3 target that represses Cd8 expression in these cells (Wang et al., 2010), and LEF1 expression was correlated with both GATA3 and Th-POK (Fig. 5 B). We found that Th-POK expression was reduced in the absence of LEF1, particularly in the CD8-expressing iNKT cells (Fig. 6 E). Therefore, similar to GATA3, LEF1 was required for proper expression of Th-POK and promoted the development of CD4+ cells while preventing the expression of CD8 on peripheral iNKT cells.


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 prevented CD8α expression on peripheral iNKT cells. (A) CD4 and CD8 expression on Tetr+TCRβ+ iNKT cells from the liver of control or Lef1Δ/Δ mice was determined by flow cytometry. (B) The mean number of CD4+, DN, and CD8+ iNKT cells from the indicated tissues in control and Lef1Δ/Δ mice. Data were averaged from at least six independent experiments. (C) CD45.1+CD45.2+ control and CD45.1−CD45.2+Lef1Δ/Δ FACS-sorted LK bone marrow cells were mixed at 1:1 ratio and transferred into lethally irradiated CD45.1+ C57BL/6 mice. 6–8 wk later, reconstituted mice were analyzed by flow cytometry for CD4 and CD8 expression on control or Lef1Δ/Δ iNKT cells from the liver. (D) The mean percentage of the CD4+, DN, and CD8+ liver iNKT cells from competitive BM chimeras. Data are representative of three independent experiments with two to three chimeras each. (E) Th-POK and CD8α expression in splenic iNKT cells from the indicated mice, as determined by flow cytometry. Data are representative of five independent experiments with one to two mice per genotype. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
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fig6: LEF1 prevented CD8α expression on peripheral iNKT cells. (A) CD4 and CD8 expression on Tetr+TCRβ+ iNKT cells from the liver of control or Lef1Δ/Δ mice was determined by flow cytometry. (B) The mean number of CD4+, DN, and CD8+ iNKT cells from the indicated tissues in control and Lef1Δ/Δ mice. Data were averaged from at least six independent experiments. (C) CD45.1+CD45.2+ control and CD45.1−CD45.2+Lef1Δ/Δ FACS-sorted LK bone marrow cells were mixed at 1:1 ratio and transferred into lethally irradiated CD45.1+ C57BL/6 mice. 6–8 wk later, reconstituted mice were analyzed by flow cytometry for CD4 and CD8 expression on control or Lef1Δ/Δ iNKT cells from the liver. (D) The mean percentage of the CD4+, DN, and CD8+ liver iNKT cells from competitive BM chimeras. Data are representative of three independent experiments with two to three chimeras each. (E) Th-POK and CD8α expression in splenic iNKT cells from the indicated mice, as determined by flow cytometry. Data are representative of five independent experiments with one to two mice per genotype. All error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Mentions: In the liver and spleen of Lef1Δ/Δ mice, the frequency of CD4+ iNKT cells was reduced and a novel population of CD8+ iNKT cells emerged (Fig. 6, A and B). The loss of CD4 and increased expression of CD8 on liver iNKT cells was intrinsic to the Lef1Δ/Δ cells as determined in mixed bone marrow chimeras (Fig. 6, C and D). We examined the expression of Th-POK in Lef1Δ/Δ splenic iNKT cells because Th-POK has been implicated as a GATA3 target that represses Cd8 expression in these cells (Wang et al., 2010), and LEF1 expression was correlated with both GATA3 and Th-POK (Fig. 5 B). We found that Th-POK expression was reduced in the absence of LEF1, particularly in the CD8-expressing iNKT cells (Fig. 6 E). Therefore, similar to GATA3, LEF1 was required for proper expression of Th-POK and promoted the development of CD4+ cells while preventing the expression of CD8 on peripheral iNKT cells.

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