Figure 7
Effects of recombinant thioredoxin (hr-Trx) on cytokine production in primary immune cells. A. Thioredoxin modulates IFN-γ- vs. IL-4-producing CD4+ T cell populations. Human peripheral lymphocytes (1 × 106) were treated with media alone or with human recombinant thioredoxin (hr-Trx) for 2 or 16 h. The cells were then stimulated with PMA (30 ng/ml) + ionomycin (1 μg/ml) in the presence of brefeldin A (10 μg/ml) for 5 h, after which they were permeabilized and stained with FITC-conjugated anti-human IFN-γ antibodies and PE-conjugated anti-human IL-4 antibodies. These cells were then restained with PerCP-conjugated anti-human CD4 antibodies to enable separation of the Th cells. FACS analysis was performed to determine the intracellular cytokine levels. B. Thioredoxin increases the IFN-γ mRNA levels in human PBMCs. Human PBMCs (1 × 106) were treated with media alone or with hr-Trx for 16 h. The cells were then stimulated with PMA (20 ng/ml) + ionomycin (0.5 μg/ml) for 5 h. The total RNA was then isolated and analyzed by real time RT-PCR using primers specific for IFN-γ. IFN-γ mRNA expression levels were normalized with the internal control, GAPDH.