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

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY

卷:126

Article

Lannigan, Josephine A.²  Solga, Michael D.²  Patrie, James³  Woodfolk, Judith A.¹ 

[1] Univ Virginia Hlth Syst, Div Allergy, Asthma & Allerg Dis Ctr, Charlottesville, VA 22908 USA

[2] Univ Virginia Hlth Syst, Dept Microbiol, Charlottesville, VA 22908 USA

[3] Univ Virginia Hlth Syst, Dept Publ Hlth Sci, Charlottesville, VA 22908 USA

关键词: TSLP;    TSLP receptor;    atopic dermatitis;    T(H)2 cells;    flow cytometry imaging;   

DOI  :  10.1016/j.jaci.2010.07.023

来源: Elsevier

【 摘 要 】

Background: Thymic stromal lymphopoietin (TSLP) is expressed at sites of allergic inflammation, including eczematous skin. This cytokine has been reported to exert its T(H)2-inducing properties through dendritic cells. Expression of TSLP receptor on the surface of activated T(H)2 cells could amplify T(H)2 responses at inflamed sites through the direct actions of TSLP. Objective: To test rigorously whether T(H)2 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(H)2-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: T(H)2-promoting stimuli induced a robust population of activated T(H)2 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(H)2 cells. The direct action of TSLP on T cells could amplify T(H)2 responses at

【 授权许可】

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