BCL-2 protects human and mouse Th17 cells from glucocorticoid-induced apoptosis

J. Banuelos, S. Shin, Y. Cao, B. S. Bochner, L. Morales-Nebreda, G. R S Budinger, L. Zhou, S. Li, J. Xin, M. W. Lingen, C. Dong, R. P. Schleimer, N. Z. Lu

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Background Glucocorticoid resistance has been associated with Th17-driven inflammation, the mechanisms of which are not clear. We determined whether human and mouse Th17 cells are resistant to glucocorticoid-induced apoptosis. Methods Freshly isolated human blood Th17 cells and in vitro differentiated Th17 cells from IL-17F red fluorescent protein reporter mice were treated with dexamethasone, a potent glucocorticoid. Apoptosis was measured using annexin V and DAPI staining. Screening of apoptosis genes was performed using the apoptosis PCR array. Levels of molecules involved in apoptosis were measured using quantitative RT-PCR, flow cytometry, and Western blotting. Knockdown of BCL-2 in murine Th17 cells was performed via retroviral transduction. Cytokines were measured using ELISA. A murine Th17-driven severe asthma model was examined for Th17 glucocorticoid sensitivity in vivo. Results Human and mouse Th17 cells and mouse Th2 cells were resistant to glucocorticoid-induced apoptosis. Th17 cells had glucocorticoid receptors levels comparable to those in other T effectors cells. Th17 cells had high levels of BCL-2, knockdown of which sensitized Th17 cells to dexamethasone-induced apoptosis. Production of IL-22, but not IL-17A and IL-17F, was suppressed by glucocorticoids. STAT3 phosphorylation in Th17 cells was insensitive to glucocorticoid inhibition. Lung Th17 cells in the murine severe asthma model were enhanced, rather than suppressed, by glucocorticoids. Conclusion Th17 cells are resistant to glucocorticoid-induced apoptosis and cytokine suppression, at least in part due to high levels of BCL-2. These findings support a role of Th17 cells in glucocorticoid-resistant inflammatory conditions such as certain endotypes of asthma.

Original languageEnglish (US)
Pages (from-to)640-650
Number of pages11
JournalAllergy: European Journal of Allergy and Clinical Immunology
Volume71
Issue number5
DOIs
StatePublished - May 1 2016

Fingerprint

Th17 Cells
Glucocorticoids
Apoptosis
Interleukin-17
Asthma
Dexamethasone
Cytokines
Polymerase Chain Reaction
Th2 Cells
Annexin A5
Glucocorticoid Receptors
Blood Cells
Flow Cytometry

Keywords

  • T cells
  • Th17 cells
  • apoptosis
  • asthma
  • glucocorticoids

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Banuelos, J. ; Shin, S. ; Cao, Y. ; Bochner, B. S. ; Morales-Nebreda, L. ; Budinger, G. R S ; Zhou, L. ; Li, S. ; Xin, J. ; Lingen, M. W. ; Dong, C. ; Schleimer, R. P. ; Lu, N. Z. / BCL-2 protects human and mouse Th17 cells from glucocorticoid-induced apoptosis. In: Allergy: European Journal of Allergy and Clinical Immunology. 2016 ; Vol. 71, No. 5. pp. 640-650.
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abstract = "Background Glucocorticoid resistance has been associated with Th17-driven inflammation, the mechanisms of which are not clear. We determined whether human and mouse Th17 cells are resistant to glucocorticoid-induced apoptosis. Methods Freshly isolated human blood Th17 cells and in vitro differentiated Th17 cells from IL-17F red fluorescent protein reporter mice were treated with dexamethasone, a potent glucocorticoid. Apoptosis was measured using annexin V and DAPI staining. Screening of apoptosis genes was performed using the apoptosis PCR array. Levels of molecules involved in apoptosis were measured using quantitative RT-PCR, flow cytometry, and Western blotting. Knockdown of BCL-2 in murine Th17 cells was performed via retroviral transduction. Cytokines were measured using ELISA. A murine Th17-driven severe asthma model was examined for Th17 glucocorticoid sensitivity in vivo. Results Human and mouse Th17 cells and mouse Th2 cells were resistant to glucocorticoid-induced apoptosis. Th17 cells had glucocorticoid receptors levels comparable to those in other T effectors cells. Th17 cells had high levels of BCL-2, knockdown of which sensitized Th17 cells to dexamethasone-induced apoptosis. Production of IL-22, but not IL-17A and IL-17F, was suppressed by glucocorticoids. STAT3 phosphorylation in Th17 cells was insensitive to glucocorticoid inhibition. Lung Th17 cells in the murine severe asthma model were enhanced, rather than suppressed, by glucocorticoids. Conclusion Th17 cells are resistant to glucocorticoid-induced apoptosis and cytokine suppression, at least in part due to high levels of BCL-2. These findings support a role of Th17 cells in glucocorticoid-resistant inflammatory conditions such as certain endotypes of asthma.",
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author = "J. Banuelos and S. Shin and Y. Cao and Bochner, {B. S.} and L. Morales-Nebreda and Budinger, {G. R S} and L. Zhou and S. Li and J. Xin and Lingen, {M. W.} and C. Dong and Schleimer, {R. P.} and Lu, {N. Z.}",
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BCL-2 protects human and mouse Th17 cells from glucocorticoid-induced apoptosis. / Banuelos, J.; Shin, S.; Cao, Y.; Bochner, B. S.; Morales-Nebreda, L.; Budinger, G. R S; Zhou, L.; Li, S.; Xin, J.; Lingen, M. W.; Dong, C.; Schleimer, R. P.; Lu, N. Z.

In: Allergy: European Journal of Allergy and Clinical Immunology, Vol. 71, No. 5, 01.05.2016, p. 640-650.

Research output: Contribution to journalArticle

TY - JOUR

T1 - BCL-2 protects human and mouse Th17 cells from glucocorticoid-induced apoptosis

AU - Banuelos, J.

AU - Shin, S.

AU - Cao, Y.

AU - Bochner, B. S.

AU - Morales-Nebreda, L.

AU - Budinger, G. R S

AU - Zhou, L.

AU - Li, S.

AU - Xin, J.

AU - Lingen, M. W.

AU - Dong, C.

AU - Schleimer, R. P.

AU - Lu, N. Z.

PY - 2016/5/1

Y1 - 2016/5/1

N2 - Background Glucocorticoid resistance has been associated with Th17-driven inflammation, the mechanisms of which are not clear. We determined whether human and mouse Th17 cells are resistant to glucocorticoid-induced apoptosis. Methods Freshly isolated human blood Th17 cells and in vitro differentiated Th17 cells from IL-17F red fluorescent protein reporter mice were treated with dexamethasone, a potent glucocorticoid. Apoptosis was measured using annexin V and DAPI staining. Screening of apoptosis genes was performed using the apoptosis PCR array. Levels of molecules involved in apoptosis were measured using quantitative RT-PCR, flow cytometry, and Western blotting. Knockdown of BCL-2 in murine Th17 cells was performed via retroviral transduction. Cytokines were measured using ELISA. A murine Th17-driven severe asthma model was examined for Th17 glucocorticoid sensitivity in vivo. Results Human and mouse Th17 cells and mouse Th2 cells were resistant to glucocorticoid-induced apoptosis. Th17 cells had glucocorticoid receptors levels comparable to those in other T effectors cells. Th17 cells had high levels of BCL-2, knockdown of which sensitized Th17 cells to dexamethasone-induced apoptosis. Production of IL-22, but not IL-17A and IL-17F, was suppressed by glucocorticoids. STAT3 phosphorylation in Th17 cells was insensitive to glucocorticoid inhibition. Lung Th17 cells in the murine severe asthma model were enhanced, rather than suppressed, by glucocorticoids. Conclusion Th17 cells are resistant to glucocorticoid-induced apoptosis and cytokine suppression, at least in part due to high levels of BCL-2. These findings support a role of Th17 cells in glucocorticoid-resistant inflammatory conditions such as certain endotypes of asthma.

AB - Background Glucocorticoid resistance has been associated with Th17-driven inflammation, the mechanisms of which are not clear. We determined whether human and mouse Th17 cells are resistant to glucocorticoid-induced apoptosis. Methods Freshly isolated human blood Th17 cells and in vitro differentiated Th17 cells from IL-17F red fluorescent protein reporter mice were treated with dexamethasone, a potent glucocorticoid. Apoptosis was measured using annexin V and DAPI staining. Screening of apoptosis genes was performed using the apoptosis PCR array. Levels of molecules involved in apoptosis were measured using quantitative RT-PCR, flow cytometry, and Western blotting. Knockdown of BCL-2 in murine Th17 cells was performed via retroviral transduction. Cytokines were measured using ELISA. A murine Th17-driven severe asthma model was examined for Th17 glucocorticoid sensitivity in vivo. Results Human and mouse Th17 cells and mouse Th2 cells were resistant to glucocorticoid-induced apoptosis. Th17 cells had glucocorticoid receptors levels comparable to those in other T effectors cells. Th17 cells had high levels of BCL-2, knockdown of which sensitized Th17 cells to dexamethasone-induced apoptosis. Production of IL-22, but not IL-17A and IL-17F, was suppressed by glucocorticoids. STAT3 phosphorylation in Th17 cells was insensitive to glucocorticoid inhibition. Lung Th17 cells in the murine severe asthma model were enhanced, rather than suppressed, by glucocorticoids. Conclusion Th17 cells are resistant to glucocorticoid-induced apoptosis and cytokine suppression, at least in part due to high levels of BCL-2. These findings support a role of Th17 cells in glucocorticoid-resistant inflammatory conditions such as certain endotypes of asthma.

KW - T cells

KW - Th17 cells

KW - apoptosis

KW - asthma

KW - glucocorticoids

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U2 - 10.1111/all.12840

DO - 10.1111/all.12840

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JO - Allergy: European Journal of Allergy and Clinical Immunology

JF - Allergy: European Journal of Allergy and Clinical Immunology

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