Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a "proallergic" milieu

Amanda J. Reefer, Kathryn E. Hulse, Josephine A. Lannigan, Michael D. Solga, Paul W. Wright, Libby A. Kelly, James Patrie, Martin D. Chapman, Judith A. Woodfolk

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Background: Thymic stromal lymphopoietin (TSLP) is expressed at sites of allergic inflammation, including eczematous skin. This cytokine has been reported to exert its TH2-inducing properties through dendritic cells. Expression of TSLP receptor on the surface of activated TH2 cells could amplify TH2 responses at inflamed sites through the direct actions of TSLP. Objective: To test rigorously whether TH2 cells induced by "proallergic" factors express TSLP receptor and characterize these cells using an experimental platform that combines flow cytometry with microscopic capabilities. Methods: CD4+ T cells isolated from patients with atopic dermatitis or normal healthy controls were cocultured with autologous dendritic cells in the presence of T H2-promoting stimuli (TSLP ± allergen and staphylococcal enterotoxin B ± TSLP). Surface expression of TSLP receptor was analyzed by image-based flow cytometry, and responsiveness of purified T cells to TSLP was assessed by phosphorylation of signal transducer and activator of transcription-5 and cytokine secretion. Results: TH2-promoting stimuli induced a robust population of activated TH2 cells (CD25 +IL-4+). Regardless of the nature of the stimulus, flow cytometry imaging confirmed that T cells expressing TSLP receptor were rare, constituting a minor fraction of the IL-4+ T cell pool; however, TSLP responsiveness was nonetheless detectable. Analysis of cell size and nuclear morphology revealed preferential expression of TSLP receptor on IL-4-expressing cells undergoing mitosis. Analysis of lesional skin in atopic dermatitis supported the view that rare IL-4+ T cells expressing TSLP receptor are present at inflamed sites. Conclusion: In a "proallergic" milieu, TSLP receptor is preferentially expressed on rare actively dividing T H2 cells. The direct action of TSLP on T cells could amplify T H2 responses at sites of allergic inflammation.

Original languageEnglish (US)
JournalJournal of Allergy and Clinical Immunology
Volume126
Issue number5
DOIs
StatePublished - Jan 1 2010

Fingerprint

Flow Cytometry
T-Lymphocytes
Interleukin-4
Atopic Dermatitis
thymic stromal lymphopoietin
Dendritic Cells
Cell Nucleus Size
STAT5 Transcription Factor
Interleukin-4 Receptors
Cytokines
Inflammation
Skin
Mitosis
Allergens
Phosphorylation

Keywords

  • Atopic dermatitis
  • Flow cytometry imaging
  • TSLP
  • TSLP receptor

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Reefer, Amanda J. ; Hulse, Kathryn E. ; Lannigan, Josephine A. ; Solga, Michael D. ; Wright, Paul W. ; Kelly, Libby A. ; Patrie, James ; Chapman, Martin D. ; Woodfolk, Judith A. / Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a "proallergic" milieu. In: Journal of Allergy and Clinical Immunology. 2010 ; Vol. 126, No. 5.
@article{9b875ee469cc4ae799a4c128129dc32e,
title = "Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a {"}proallergic{"} milieu",
abstract = "Background: Thymic stromal lymphopoietin (TSLP) is expressed at sites of allergic inflammation, including eczematous skin. This cytokine has been reported to exert its TH2-inducing properties through dendritic cells. Expression of TSLP receptor on the surface of activated TH2 cells could amplify TH2 responses at inflamed sites through the direct actions of TSLP. Objective: To test rigorously whether TH2 cells induced by {"}proallergic{"} factors express TSLP receptor and characterize these cells using an experimental platform that combines flow cytometry with microscopic capabilities. Methods: CD4+ T cells isolated from patients with atopic dermatitis or normal healthy controls were cocultured with autologous dendritic cells in the presence of T H2-promoting stimuli (TSLP ± allergen and staphylococcal enterotoxin B ± TSLP). Surface expression of TSLP receptor was analyzed by image-based flow cytometry, and responsiveness of purified T cells to TSLP was assessed by phosphorylation of signal transducer and activator of transcription-5 and cytokine secretion. Results: TH2-promoting stimuli induced a robust population of activated TH2 cells (CD25 +IL-4+). Regardless of the nature of the stimulus, flow cytometry imaging confirmed that T cells expressing TSLP receptor were rare, constituting a minor fraction of the IL-4+ T cell pool; however, TSLP responsiveness was nonetheless detectable. Analysis of cell size and nuclear morphology revealed preferential expression of TSLP receptor on IL-4-expressing cells undergoing mitosis. Analysis of lesional skin in atopic dermatitis supported the view that rare IL-4+ T cells expressing TSLP receptor are present at inflamed sites. Conclusion: In a {"}proallergic{"} milieu, TSLP receptor is preferentially expressed on rare actively dividing T H2 cells. The direct action of TSLP on T cells could amplify T H2 responses at sites of allergic inflammation.",
keywords = "Atopic dermatitis, Flow cytometry imaging, TSLP, TSLP receptor",
author = "Reefer, {Amanda J.} and Hulse, {Kathryn E.} and Lannigan, {Josephine A.} and Solga, {Michael D.} and Wright, {Paul W.} and Kelly, {Libby A.} and James Patrie and Chapman, {Martin D.} and Woodfolk, {Judith A.}",
year = "2010",
month = "1",
day = "1",
doi = "10.1016/j.jaci.2010.07.023",
language = "English (US)",
volume = "126",
journal = "Journal of Allergy and Clinical Immunology",
issn = "0091-6749",
publisher = "Mosby Inc.",
number = "5",

}

Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a "proallergic" milieu. / Reefer, Amanda J.; Hulse, Kathryn E.; Lannigan, Josephine A.; Solga, Michael D.; Wright, Paul W.; Kelly, Libby A.; Patrie, James; Chapman, Martin D.; Woodfolk, Judith A.

In: Journal of Allergy and Clinical Immunology, Vol. 126, No. 5, 01.01.2010.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a "proallergic" milieu

AU - Reefer, Amanda J.

AU - Hulse, Kathryn E.

AU - Lannigan, Josephine A.

AU - Solga, Michael D.

AU - Wright, Paul W.

AU - Kelly, Libby A.

AU - Patrie, James

AU - Chapman, Martin D.

AU - Woodfolk, Judith A.

PY - 2010/1/1

Y1 - 2010/1/1

N2 - Background: Thymic stromal lymphopoietin (TSLP) is expressed at sites of allergic inflammation, including eczematous skin. This cytokine has been reported to exert its TH2-inducing properties through dendritic cells. Expression of TSLP receptor on the surface of activated TH2 cells could amplify TH2 responses at inflamed sites through the direct actions of TSLP. Objective: To test rigorously whether TH2 cells induced by "proallergic" factors express TSLP receptor and characterize these cells using an experimental platform that combines flow cytometry with microscopic capabilities. Methods: CD4+ T cells isolated from patients with atopic dermatitis or normal healthy controls were cocultured with autologous dendritic cells in the presence of T H2-promoting stimuli (TSLP ± allergen and staphylococcal enterotoxin B ± TSLP). Surface expression of TSLP receptor was analyzed by image-based flow cytometry, and responsiveness of purified T cells to TSLP was assessed by phosphorylation of signal transducer and activator of transcription-5 and cytokine secretion. Results: TH2-promoting stimuli induced a robust population of activated TH2 cells (CD25 +IL-4+). Regardless of the nature of the stimulus, flow cytometry imaging confirmed that T cells expressing TSLP receptor were rare, constituting a minor fraction of the IL-4+ T cell pool; however, TSLP responsiveness was nonetheless detectable. Analysis of cell size and nuclear morphology revealed preferential expression of TSLP receptor on IL-4-expressing cells undergoing mitosis. Analysis of lesional skin in atopic dermatitis supported the view that rare IL-4+ T cells expressing TSLP receptor are present at inflamed sites. Conclusion: In a "proallergic" milieu, TSLP receptor is preferentially expressed on rare actively dividing T H2 cells. The direct action of TSLP on T cells could amplify T H2 responses at sites of allergic inflammation.

AB - Background: Thymic stromal lymphopoietin (TSLP) is expressed at sites of allergic inflammation, including eczematous skin. This cytokine has been reported to exert its TH2-inducing properties through dendritic cells. Expression of TSLP receptor on the surface of activated TH2 cells could amplify TH2 responses at inflamed sites through the direct actions of TSLP. Objective: To test rigorously whether TH2 cells induced by "proallergic" factors express TSLP receptor and characterize these cells using an experimental platform that combines flow cytometry with microscopic capabilities. Methods: CD4+ T cells isolated from patients with atopic dermatitis or normal healthy controls were cocultured with autologous dendritic cells in the presence of T H2-promoting stimuli (TSLP ± allergen and staphylococcal enterotoxin B ± TSLP). Surface expression of TSLP receptor was analyzed by image-based flow cytometry, and responsiveness of purified T cells to TSLP was assessed by phosphorylation of signal transducer and activator of transcription-5 and cytokine secretion. Results: TH2-promoting stimuli induced a robust population of activated TH2 cells (CD25 +IL-4+). Regardless of the nature of the stimulus, flow cytometry imaging confirmed that T cells expressing TSLP receptor were rare, constituting a minor fraction of the IL-4+ T cell pool; however, TSLP responsiveness was nonetheless detectable. Analysis of cell size and nuclear morphology revealed preferential expression of TSLP receptor on IL-4-expressing cells undergoing mitosis. Analysis of lesional skin in atopic dermatitis supported the view that rare IL-4+ T cells expressing TSLP receptor are present at inflamed sites. Conclusion: In a "proallergic" milieu, TSLP receptor is preferentially expressed on rare actively dividing T H2 cells. The direct action of TSLP on T cells could amplify T H2 responses at sites of allergic inflammation.

KW - Atopic dermatitis

KW - Flow cytometry imaging

KW - TSLP

KW - TSLP receptor

UR - http://www.scopus.com/inward/record.url?scp=78049465932&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78049465932&partnerID=8YFLogxK

U2 - 10.1016/j.jaci.2010.07.023

DO - 10.1016/j.jaci.2010.07.023

M3 - Article

VL - 126

JO - Journal of Allergy and Clinical Immunology

JF - Journal of Allergy and Clinical Immunology

SN - 0091-6749

IS - 5

ER -