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CD4 dim CD25 bright Treg cell frequencies above a standardized gating threshold are similar in asthmatics and controls
Author(s) -
Hoffmann Hans Jürgen,
Malling Tine M.,
Topcu Ayfer,
Ryder Lars P.,
Nielsen Kaspar R.,
Varming Kim,
Dahl Ronald,
Omland Øyvind,
Sigsgaard Torben
Publication year - 2007
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20389
Subject(s) - il 2 receptor , interleukin 7 receptor , foxp3 , flow cytometry , immunology , population , medicine , immune system , biology , t cell , environmental health
Background: Thymus selected CD4 + CD25 bright natural regulatory Treg cells expressing FOXP3 may contribute to control of immune responses. No unique markers have been available to identify and characterize Treg. We present a gating strategy that allows enumeration of Treg on the basis of CD4 and CD25 and investigate whether asthmatics have fewer Treg than controls. Methods: Asthmatics and controls were selected from responses to a mailed questionnaire. CD25, CD4, HLA DR, and appropriate isotypes were recorded by flow cytometry. Results: The CD4 T cells expressing most CD25 are a separate population expressing FOXP3 and lower levels of CD4 and CD127. On a CD4 CD25 dot‐plot, the CD4 MFI of Treg for 152 participants was calculated to be 0.83 ± 0.043*MFI of CD25 bright T‐cells. CD4 dim CD25 bright T cells in a rectangular gate with a CD4 MFI ≤ 0.9 (0.83 + [2*0.043])*MFI of CD25 + T cells were enumerated and shown to be similar for controls (median 8.34%) and asthmatics (median 10.1%). HLA DR expression on Treg correlated with CD25 expression. Conclusions: A standardized two color gating method defines Treg. It may be applied in most clinical scenarios and is useful for sorting viable Treg. Asthmatics and controls have similar numbers of Treg. © 2007 International Society for Analytical Cytology