Table 1.
Expression profiles of key T cell lineage transcription factors in CD46-induced effector (TH1) and suppressor (Treg) cell populations
Transcription factor | IFN-γ+IL-10−(TH1) | IFN-γ+IL-10+ (Treg) | IFN-γ−IL-10+ (Treg) | TH1a | TH2a |
---|---|---|---|---|---|
T-bet | +++ | +++ | +++ | ++++ | − |
GATA-3b | −/+ | + | + | −/+ | ++++ |
Foxp3 | − | − | − | − | − |
p-STAT4 | ++ | +++ | ++ | ++++ | ++ |
p-STAT5 | +++ | +++ | + | ++++ | ++ |
p-STAT6 | + | + | −/+ | + | ++++ |
p-Erk | ++ | +++ | +++ | ++++ | ++ |
ICER/CREM | +++ | ++ | + | ++ | ++ |
IL-10- and IFN-γ-secretion assays plus intracellular flow cytometry staining for T cell lineage markers in purified CD4+ T cells activated for 36 h with anti-CD3 and anti-CD46 in a high concentration of IL-2; transcription factor expression is presented relative to the expression in TH1 or TH2 cells as follows: ++++, highest; +++, strong; ++, moderate; +, low; −/+, weak but detectable; −, undetectable; p-, phosphorylated.
Purity of TH1 and TH2 cells assessed as percent IFN-γ+IL-4- cells (TH1) or IFN-γ−IL-4+ cells (TH2) after restimulation with anti-CD3 and anti-CD28 (>85% in all assays).
GATA-3 mRNA assessed by quantitative real-time RT-PCR analysis because of the lack of a suitable antibody.