Figure 3
Unstimulated CD4+T lymphocytes release IL-2 and TNF-α when treated with exosomes from F12/Hut-78 cells. A. Detection of IL-2-producing cells. 2 × 105 unstimulated CD4+ T lymphocytes were challenged with 60 μU of exosomes from either Hut-78 or F12/Hut-78 cells, or mock-challenged (Nil), and then incubated for 48 hours in ELISPOT microwells previously coated with an anti-IL-2 monoclonal antibody. Shown are the mean number + SD of IL-2 spot forming units (SFU)/106 cells calculated from five independent experiments. *p < 0.05. B. Detection of TNF-α on supernatants. 105 unstimulated CD4+ T lymphocytes were challenged with increasing amounts (i.e., from 15 to 60 μU) of exosomes from Hut-78 or F12/Hut-78 cells, or mock-challenged (Nil). As control, cells were also treated with 2 μg/mL of PHA. After extensive washings, the cells were seeded in complete medium, and supernatants were harvested 6 hours later. TNF-α contents were determined by ELISA. Shown are the mean concentrations + SD as calculated from three independent experiments. Nd: not detectable. *p < 0.05.