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 journalArticlepeer-review

25 Scopus citations

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)
Pages (from-to)1049-1058.e10
JournalJournal of Allergy and Clinical Immunology
Volume126
Issue number5
DOIs
StatePublished - Nov 2010

Keywords

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

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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