Skip to main content
. 2017 Sep 21;36(20):3012–3028. doi: 10.15252/embj.201696003

Figure 5. Comparison of the effect of DC‐derived EVs versus the whole DC on T cells.

Figure 5

  1. Total CD4+ T cells were isolated, stained with Cell Trace Violet dye and cultured for 6 days with increasing numbers of in vitro differentiated DCs or with EVs purified from these DCs (2K, 10K and 100K). Proliferation was calculated by dilution of the fluorescent dye on CD3+CD4+ cells (n = 8, one symbol per individual DC‐EV:T‐cell combination). Red line indicates median.
  2. The concentration of IL‐13 and IFN‐γ was measured in culture supernatant after 6 days of culture of total CD4+ T cell with 125 and 250 whole DCs or with their secreted EVs. Results are expressed as fold induction compared to untreated CD4+ T cells (n = 8, one symbol per individual DC‐EV:T‐cell combination). Red line indicates median. The range of cytokines secreted by untreated CD4+ T cells was 20.8–402.7 pg/ml of IL‐13; 24.8–143.6 pg/ml of IFN‐γ.
  3. Th1 to Th2 ratio was calculated by dividing the concentration of IFN‐γ to the concentration of IL‐13 for each donor, for the whole DC‐T‐cell co‐culture (125 and 250 DCs) and for the EV‐T‐cell culture (n = 8). Results are shown as a box plot (25th to 75th percentiles, the line represents the median; whiskers, min to max, points represent outliers as calculated by Tukey's test). *P < 0.05; **P < 0.01; ***P < 0.001 (Friedman test).
  4. DC and T cells were co‐cultured in a Transwell system, separated through either a 0.4‐μm or a 1‐μm porous membrane at a 1:1 or 1:8 DC:T‐cell ratio. After 6 days, the supernatants were analysed for the presence of IL‐13 and IFN‐γ by cytometric bead array. Experiments done with 0.4‐μm or with 1‐μm pore are completely independent (not paired donors) (n = 4–5, one symbol per individual DC:T‐cell combination). Red line indicates median. *P < 0.05, **P < 0.01; one‐way ANOVA followed by Bonferroni's multiple comparison test.