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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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Notch receptor expression and signaling in CD8+ TEFF cells. (A) Relative gene expression of Notch family members in TEFF versus TCM cells. The expression of Notch on TEFF and TCM cells was determined by gene chip analysis. The relative gene expression difference for the hybridization signal is depicted as a ratio. #, P < 0.05 summarizing the results of three separate experiments performed in duplicate. (B) Notch signaling is activated in TEFF cells. TEFF cells were stimulated with anti-CD3 and anti-CD28 or SIINFEKL for 24 h after incubation of the cells with DMSO or GSI. Cell lysates were analyzed for the expression of Notch1 by Western blotting, and β-actin was used as a loading control. One representative of three similar experiments is shown.
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fig1: Notch receptor expression and signaling in CD8+ TEFF cells. (A) Relative gene expression of Notch family members in TEFF versus TCM cells. The expression of Notch on TEFF and TCM cells was determined by gene chip analysis. The relative gene expression difference for the hybridization signal is depicted as a ratio. #, P < 0.05 summarizing the results of three separate experiments performed in duplicate. (B) Notch signaling is activated in TEFF cells. TEFF cells were stimulated with anti-CD3 and anti-CD28 or SIINFEKL for 24 h after incubation of the cells with DMSO or GSI. Cell lysates were analyzed for the expression of Notch1 by Western blotting, and β-actin was used as a loading control. One representative of three similar experiments is shown.

Mentions: We previously showed that the development of AHR and eosinophilic inflammation in CD8−/− mice was lower than in WT mice but could be fully restored after transfer of CD8+ T cells from antigen-primed donors or after transfer of in vitro–generated CD8+ TEFF cells but not CD8+ TCM cells (10). Reconstitution of heightened airway responsiveness by TEFF cells was paralleled by restoration of bronchoalveolar lavage (BAL) and tissue eosinophilia, BAL IL-13 levels, and goblet cell metaplasia. To determine if there were differences in gene expression between TEFF and TCM cells, microarray analysis was performed (the microarray data have been deposited in the GEO database under accession number GSM8632). In the analysis of TEFF and TCM cell total RNA, an up-regulation of Notch1 was detected in TEFF cells, which was >1,000-fold higher than in TCM cells (P < 0.05) (Fig. 1 A). In contrast, the expression of Notch2 and Notch3 was only minimally higher in TEFF cells.


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)

Notch receptor expression and signaling in CD8+ TEFF cells. (A) Relative gene expression of Notch family members in TEFF versus TCM cells. The expression of Notch on TEFF and TCM cells was determined by gene chip analysis. The relative gene expression difference for the hybridization signal is depicted as a ratio. #, P < 0.05 summarizing the results of three separate experiments performed in duplicate. (B) Notch signaling is activated in TEFF cells. TEFF cells were stimulated with anti-CD3 and anti-CD28 or SIINFEKL for 24 h after incubation of the cells with DMSO or GSI. Cell lysates were analyzed for the expression of Notch1 by Western blotting, and β-actin was used as a loading control. One representative of three similar experiments is shown.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2373841&req=5

fig1: Notch receptor expression and signaling in CD8+ TEFF cells. (A) Relative gene expression of Notch family members in TEFF versus TCM cells. The expression of Notch on TEFF and TCM cells was determined by gene chip analysis. The relative gene expression difference for the hybridization signal is depicted as a ratio. #, P < 0.05 summarizing the results of three separate experiments performed in duplicate. (B) Notch signaling is activated in TEFF cells. TEFF cells were stimulated with anti-CD3 and anti-CD28 or SIINFEKL for 24 h after incubation of the cells with DMSO or GSI. Cell lysates were analyzed for the expression of Notch1 by Western blotting, and β-actin was used as a loading control. One representative of three similar experiments is shown.
Mentions: We previously showed that the development of AHR and eosinophilic inflammation in CD8−/− mice was lower than in WT mice but could be fully restored after transfer of CD8+ T cells from antigen-primed donors or after transfer of in vitro–generated CD8+ TEFF cells but not CD8+ TCM cells (10). Reconstitution of heightened airway responsiveness by TEFF cells was paralleled by restoration of bronchoalveolar lavage (BAL) and tissue eosinophilia, BAL IL-13 levels, and goblet cell metaplasia. To determine if there were differences in gene expression between TEFF and TCM cells, microarray analysis was performed (the microarray data have been deposited in the GEO database under accession number GSM8632). In the analysis of TEFF and TCM cell total RNA, an up-regulation of Notch1 was detected in TEFF cells, which was >1,000-fold higher than in TCM cells (P < 0.05) (Fig. 1 A). In contrast, the expression of Notch2 and Notch3 was only minimally higher in TEFF cells.

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