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Essential role of Notch signaling in effector memory CD8+ T cell-mediated airway hyperresponsiveness and inflammation.

Okamoto M, Takeda K, Joetham A, Ohnishi H, Matsuda H, Swasey CH, Swanson BJ, Yasutomo K, Dakhama A, Gelfand EW - J. Exp. Med. (2008)

Bottom Line: Examining transcription levels, there was a strong induction of Notch1 in T(EFF) cells compared with central memory CD8(+) T cells.These effects of GSI were also associated with increased expression of the Notch ligand Delta1 in T(EFF) cells.These results demonstrate a role for Notch in skewing the T cell response from a T helper (Th)2 to a Th1 phenotype as a consequence of the inhibition of Notch receptor activation and the up-regulation of the Notch ligand Delta1.

View Article: PubMed Central - PubMed

Affiliation: Division of Cell Biology, Department of Pediatrics, National Jewish Medical and Research Center, Denver, CO 80206, USA.

ABSTRACT
Adoptive transfer of in vivo-primed CD8(+) T cells or in vitro-generated effector memory CD8(+) T (T(EFF)) cells restores airway hyperresponsiveness (AHR) and airway inflammation in CD8-deficient (CD8(-/-)) mice. Examining transcription levels, there was a strong induction of Notch1 in T(EFF) cells compared with central memory CD8(+) T cells. Treatment of T(EFF) cells with a gamma-secretase inhibitor (GSI) strongly inhibited Notch signaling in these cells, and after adoptive transfer, GSI-treated T(EFF) cells failed to restore AHR and airway inflammation in sensitized and challenged recipient CD8(-/-) mice, or to enhance these responses in recipient wild-type (WT) mice. These effects of GSI were also associated with increased expression of the Notch ligand Delta1 in T(EFF) cells. Treatment of sensitized and challenged WT mice with Delta1-Fc resulted in decreased AHR and airway inflammation accompanied by higher levels of interferon gamma in bronchoalveolar lavage fluid. These results demonstrate a role for Notch in skewing the T cell response from a T helper (Th)2 to a Th1 phenotype as a consequence of the inhibition of Notch receptor activation and the up-regulation of the Notch ligand Delta1. These data are the first to show a functional role for Notch in the challenge phase of CD8(+) T cell-mediated development of AHR and airway inflammation, and identify Delta1 as an important regulator of allergic airway inflammation.

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Transfer of GSI-TEFF cells fails to enhance lung allergic responses in WT recipients. Sensitized and challenged WT mice received GSI-TEFF or DMSO-TEFF cells before secondary challenge (OVA/OVA/OVA). Control mice were those that were sensitized and challenged but received PBS at the time of secondary challenge (OVA/OVA/PBS). (A) RL values were obtained in response to increasing concentrations of inhaled MCh, as described in Materials and methods. Data represent the mean ± SEM (n = 12 in each group). #, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice or GSI-TEFF cell recipients. (B) Cellular composition of BAL fluid. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (C) CD4+ and CD8+ T cells in BAL fluid. ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (D) Cytokine levels in BAL fluid of WT mice that received GSI-TEFF or DMSO-TEFF cells. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT control mice. (E) Relative expression levels of GATA-3 and T-bet in the lung were determined by quantitative real-time PCR. Lung cells are from the same groups as described in A. They were isolated using collagenase digestion, and RNA was prepared. Results are representative of three independent experiments. The results for each group are expressed as mean ± SEM. #, significant difference (P < 0.05) between GSI-TEFF cell–transferred mice and DMSO-TEFF cell–transferred mice or WT control mice.
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fig2: Transfer of GSI-TEFF cells fails to enhance lung allergic responses in WT recipients. Sensitized and challenged WT mice received GSI-TEFF or DMSO-TEFF cells before secondary challenge (OVA/OVA/OVA). Control mice were those that were sensitized and challenged but received PBS at the time of secondary challenge (OVA/OVA/PBS). (A) RL values were obtained in response to increasing concentrations of inhaled MCh, as described in Materials and methods. Data represent the mean ± SEM (n = 12 in each group). #, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice or GSI-TEFF cell recipients. (B) Cellular composition of BAL fluid. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (C) CD4+ and CD8+ T cells in BAL fluid. ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (D) Cytokine levels in BAL fluid of WT mice that received GSI-TEFF or DMSO-TEFF cells. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT control mice. (E) Relative expression levels of GATA-3 and T-bet in the lung were determined by quantitative real-time PCR. Lung cells are from the same groups as described in A. They were isolated using collagenase digestion, and RNA was prepared. Results are representative of three independent experiments. The results for each group are expressed as mean ± SEM. #, significant difference (P < 0.05) between GSI-TEFF cell–transferred mice and DMSO-TEFF cell–transferred mice or WT control mice.

Mentions: To assess the effects of the Notch signaling pathway on the enhancement of the functional activity of CD8+ TEFF cells in vivo, GSI-TEFF cells or DMSO-TEFF cells were transferred into sensitized and challenged recipient WT mice before secondary OVA challenge. Secondary challenge of sensitized and challenged mice led to the development of increased AHR in WT mice, illustrated by significant increases in lung resistance (RL) (Fig. 2 A), as described previously in this model (31).Fig. 2 A also illustrates the changes in RL in WT recipients of GSI-TEFF cells or DMSO-TEFF cells undergoing secondary challenge. AHR to methacholine (MCh) was significantly increased in recipients of DMSO-TEFF cells, but recipients of GSI-TEFF cells failed to increase AHR over that seen in WT mice not receiving TEFF cells. Indeed, transfer of DMSO-TEFF cells significantly increased AHR, whereas transfer of GSI-TEFF cells inhibited the response to a modest degree. In parallel to the assessment of lung function, the inflammatory cell composition of BAL fluid was examined (Fig. 2 B). Eosinophil numbers in BAL fluid were significantly increased in the DMSO-TEFF cell recipient mice, whereas GSI-TEFF cell recipient mice did not show such increases. In contrast, neutrophil numbers were increased in GSI-TEFF cell recipients. Transfer of either DMSO-TEFF cells or GSF TEFF cells increased lymphocyte numbers in BAL fluid compared with WT control mice after secondary allergen challenge.


Essential role of Notch signaling in effector memory CD8+ T cell-mediated airway hyperresponsiveness and inflammation.

Okamoto M, Takeda K, Joetham A, Ohnishi H, Matsuda H, Swasey CH, Swanson BJ, Yasutomo K, Dakhama A, Gelfand EW - J. Exp. Med. (2008)

Transfer of GSI-TEFF cells fails to enhance lung allergic responses in WT recipients. Sensitized and challenged WT mice received GSI-TEFF or DMSO-TEFF cells before secondary challenge (OVA/OVA/OVA). Control mice were those that were sensitized and challenged but received PBS at the time of secondary challenge (OVA/OVA/PBS). (A) RL values were obtained in response to increasing concentrations of inhaled MCh, as described in Materials and methods. Data represent the mean ± SEM (n = 12 in each group). #, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice or GSI-TEFF cell recipients. (B) Cellular composition of BAL fluid. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (C) CD4+ and CD8+ T cells in BAL fluid. ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (D) Cytokine levels in BAL fluid of WT mice that received GSI-TEFF or DMSO-TEFF cells. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT control mice. (E) Relative expression levels of GATA-3 and T-bet in the lung were determined by quantitative real-time PCR. Lung cells are from the same groups as described in A. They were isolated using collagenase digestion, and RNA was prepared. Results are representative of three independent experiments. The results for each group are expressed as mean ± SEM. #, significant difference (P < 0.05) between GSI-TEFF cell–transferred mice and DMSO-TEFF cell–transferred mice or WT control mice.
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fig2: Transfer of GSI-TEFF cells fails to enhance lung allergic responses in WT recipients. Sensitized and challenged WT mice received GSI-TEFF or DMSO-TEFF cells before secondary challenge (OVA/OVA/OVA). Control mice were those that were sensitized and challenged but received PBS at the time of secondary challenge (OVA/OVA/PBS). (A) RL values were obtained in response to increasing concentrations of inhaled MCh, as described in Materials and methods. Data represent the mean ± SEM (n = 12 in each group). #, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice or GSI-TEFF cell recipients. (B) Cellular composition of BAL fluid. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (C) CD4+ and CD8+ T cells in BAL fluid. ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT mice. (D) Cytokine levels in BAL fluid of WT mice that received GSI-TEFF or DMSO-TEFF cells. #, significant difference (P < 0.05) between GSI-TEFF cell recipients and DMSO-TEFF cell recipients; ‡, significant difference (P < 0.05) between DMSO-TEFF cell recipients and WT control mice; *, significant difference (P < 0.05) between GSI-TEFF cell recipients and WT control mice. (E) Relative expression levels of GATA-3 and T-bet in the lung were determined by quantitative real-time PCR. Lung cells are from the same groups as described in A. They were isolated using collagenase digestion, and RNA was prepared. Results are representative of three independent experiments. The results for each group are expressed as mean ± SEM. #, significant difference (P < 0.05) between GSI-TEFF cell–transferred mice and DMSO-TEFF cell–transferred mice or WT control mice.
Mentions: To assess the effects of the Notch signaling pathway on the enhancement of the functional activity of CD8+ TEFF cells in vivo, GSI-TEFF cells or DMSO-TEFF cells were transferred into sensitized and challenged recipient WT mice before secondary OVA challenge. Secondary challenge of sensitized and challenged mice led to the development of increased AHR in WT mice, illustrated by significant increases in lung resistance (RL) (Fig. 2 A), as described previously in this model (31).Fig. 2 A also illustrates the changes in RL in WT recipients of GSI-TEFF cells or DMSO-TEFF cells undergoing secondary challenge. AHR to methacholine (MCh) was significantly increased in recipients of DMSO-TEFF cells, but recipients of GSI-TEFF cells failed to increase AHR over that seen in WT mice not receiving TEFF cells. Indeed, transfer of DMSO-TEFF cells significantly increased AHR, whereas transfer of GSI-TEFF cells inhibited the response to a modest degree. In parallel to the assessment of lung function, the inflammatory cell composition of BAL fluid was examined (Fig. 2 B). Eosinophil numbers in BAL fluid were significantly increased in the DMSO-TEFF cell recipient mice, whereas GSI-TEFF cell recipient mice did not show such increases. In contrast, neutrophil numbers were increased in GSI-TEFF cell recipients. Transfer of either DMSO-TEFF cells or GSF TEFF cells increased lymphocyte numbers in BAL fluid compared with WT control mice after secondary allergen challenge.

Bottom Line: Examining transcription levels, there was a strong induction of Notch1 in T(EFF) cells compared with central memory CD8(+) T cells.These effects of GSI were also associated with increased expression of the Notch ligand Delta1 in T(EFF) cells.These results demonstrate a role for Notch in skewing the T cell response from a T helper (Th)2 to a Th1 phenotype as a consequence of the inhibition of Notch receptor activation and the up-regulation of the Notch ligand Delta1.

View Article: PubMed Central - PubMed

Affiliation: Division of Cell Biology, Department of Pediatrics, National Jewish Medical and Research Center, Denver, CO 80206, USA.

ABSTRACT
Adoptive transfer of in vivo-primed CD8(+) T cells or in vitro-generated effector memory CD8(+) T (T(EFF)) cells restores airway hyperresponsiveness (AHR) and airway inflammation in CD8-deficient (CD8(-/-)) mice. Examining transcription levels, there was a strong induction of Notch1 in T(EFF) cells compared with central memory CD8(+) T cells. Treatment of T(EFF) cells with a gamma-secretase inhibitor (GSI) strongly inhibited Notch signaling in these cells, and after adoptive transfer, GSI-treated T(EFF) cells failed to restore AHR and airway inflammation in sensitized and challenged recipient CD8(-/-) mice, or to enhance these responses in recipient wild-type (WT) mice. These effects of GSI were also associated with increased expression of the Notch ligand Delta1 in T(EFF) cells. Treatment of sensitized and challenged WT mice with Delta1-Fc resulted in decreased AHR and airway inflammation accompanied by higher levels of interferon gamma in bronchoalveolar lavage fluid. These results demonstrate a role for Notch in skewing the T cell response from a T helper (Th)2 to a Th1 phenotype as a consequence of the inhibition of Notch receptor activation and the up-regulation of the Notch ligand Delta1. These data are the first to show a functional role for Notch in the challenge phase of CD8(+) T cell-mediated development of AHR and airway inflammation, and identify Delta1 as an important regulator of allergic airway inflammation.

Show MeSH
Related in: MedlinePlus