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- W2073343689 abstract "RATIONALE: Because transcription factor GATA3 is required for Th2 differentiation of naive T cells, and the 7-transmembrane PGD2 receptor CRTH2 is the most selective Th2 surface marker identified to date, we hypothesized a direct correlation between the expression levels of these two proteins in polarized Th2 cultures. METHODS: Human naive CD4+ T cells were primed with anti-CD3/CD28 and cultured under conventional Th1- or Th2-polarizing conditions. After 7 days of culture, CRTH2+ cells were enriched to >70% by positive selection using immunomagnetic beads (Dynal). Surface antigen expression and single-cell cytokine gene expression in permeabilized, monensin-treated cells were analyzed by flow cytometry. GATA3 expression was analyzed by RT-PCR. RESULTS: On day 7, CRTH2 surface expression was preferentially induced in Th2-polarized cultures (mean±SE: 16.8±1.8% vs 0.5±0.1% in Th1 cells in 8 independent experiments; p<0.05). Conversely, CCR5, a Th1 surface marker, was expressed on a higher proportion of Th1 than Th2 cells (23.0±1.9% vs 0.8±0.6%; p<0.05). After stimulation with PMA and ionomycin the totality of CRTH2+ cells expressed IL-4 but not interferon-γ. As reported, GATA3 mRNA was detected at higher levels in Th2 than in Th1 cells. However, within Th2 cells, there were no differences in GATA3 mRNA expression between CRTH2-enriched and CRTH2-depleted fractions. CONCLUSIONS: Our data show that, while increased expression of GATA3 is consistently observed in differentiating Th2 cultures, fully polarized cells, identified by CRTH2 and IL-4 detection, do not express higher levels of the factor. This suggests that GATA3 is necessary but not sufficient for stable, inheritable activation of these Th2 genes." @default.
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- W2073343689 date "2005-02-01" @default.
- W2073343689 modified "2023-09-27" @default.
- W2073343689 title "Polarized expression of GATA3 is not sufficient for CRTH2 and IL-4 gene activation in differentiated human Th2 clones" @default.
- W2073343689 doi "https://doi.org/10.1016/j.jaci.2004.12.551" @default.
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