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Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture
Published on: April 8, 2016
Skin-homing CD4+ Foxp3+ T cells exert Th2-like function after staphylococcal superantigen stimulation in atopic
1Department of Pediatrics, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
Background:
The effect of staphylococcal superantigens (SsAgs) on cutaneous lymphocyte-associated antigen (CLA)(+) CD4(+) Foxp3(+) T cells of atopic dermatitis (AD) patients is unknown.
Objective:
To compare the effects of SsAgs on the ratio, function, and apoptosis of CCR6(+) subtype and CCR6(-) subtype of CLA(+) CD4(+) Foxp3(+) T cells among AD patients, asthma/allergic rhinitis (AR) patients without AD, and healthy subjects.
Methods:
Using immunofluorescence staining followed by flow cytometric analysis, we analysed peripheral blood mononuclear cells cultured with or without staphylococcal enterotoxin B (SEB) stimulation in 20 AD patients, 20 asthma/AR patients without AD, and 20 healthy subjects.
Results:
SEB decreased CCR6(+) /CCR6(-) ratio in CLA(+) CD4(+) Foxp3(+) T cells from AD patients and increased CCR6(+) /CCR6(-) ratio in those from healthy subjects. SEB induced the production of type 2 T helper cell (Th2) cytokine interleukin (IL)-5 in CCR6(-) subtype and anti-inflammatory cytokine IL-10 in CCR6(+) subtype of CLA(+) CD4(+) Foxp3(+) T cells. CLA(+) CD4(+) Foxp3(+) T cells from AD patients produced more IL-5 and less IL-10 after SEB stimulation than those from healthy subjects. CCR6(-) subtype of CLA(+) CD4(+) Foxp3(+) T cells from AD patients and CCR6(+) subtype of those cells from healthy subjects were more resistant to SEB-induced caspase-3 activation than the other subtype and those from other subjects.
Conclusions And Clinical Relevance:
Despite a phenotype of regulatory T cells, skin-homing CD4(+) Foxp3(+) T cells of AD patients exert effector Th2-like function after SsAgs stimulation, which may aggravate allergic skin inflammation.
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