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• W2136918536 abstract "BackgroundThymic 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.ObjectiveTo 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.MethodsCD4+ T cells isolated from patients with atopic dermatitis or normal healthy controls were cocultured with autologous dendritic cells in the presence of TH2-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.ResultsTH2-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.ConclusionIn a “proallergic” milieu, TSLP receptor is preferentially expressed on rare actively dividing TH2 cells. The direct action of TSLP on T cells could amplify TH2 responses at sites of allergic inflammation. 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. 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. CD4+ T cells isolated from patients with atopic dermatitis or normal healthy controls were cocultured with autologous dendritic cells in the presence of TH2-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. 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. In a “proallergic” milieu, TSLP receptor is preferentially expressed on rare actively dividing TH2 cells. The direct action of TSLP on T cells could amplify TH2 responses at sites of allergic inflammation." @default.
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• W2136918536 date "2010-11-01" @default.
• W2136918536 modified "2023-10-02" @default.
• W2136918536 title "Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a “proallergic” milieu" @default.
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• W2136918536 doi "https://doi.org/10.1016/j.jaci.2010.07.023" @default.
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